Cognitive Behavioural Therapy (CBT) Case Study Example | Topics and Well Written Essays - 3000 words

Intellectual Behavioral Therapy (CBT) - Case Study Example Some data on Karen accessible to Dr. Banks before embraced psychotherapy is sketched out underneath. Karen left her parental home at 18 years old in the wake of moving on from secondary school and moved to lodgings in the neighboring district. She looked for some kind of employment as a server at a nearby coffee shop. Only half a month subsequent to looking for some kind of employment, she wedded George whom she had met while at work. An exceptionally controlling and savage man, George ruled Karen in what appeared to be a reiteration of the conditions under which Karen grew up. Her dad had been vicious and explicitly oppressive from the time she was six years of age. As she developed more established (13) her most seasoned sibling proceeded with the sexual maltreatment, until her other sibling, closer to her in age, shut down it. She was 16 at that point.  George, her significant other, would be pleasant to her on uncommon events, taking her out to supper and moves, and would praise her on her attractive features. George end up being a desirous spouse and constrained her to surrender work. She was practically kept detainee bound to the house. Indeed, even with the incidental beatings, and not knowing any better, she set up with it and seemed to acknowledge the circumstance as ‘normal’. Sadly, George passed on in a mishap scarcely three years into their marriage. Karen was crushed. She had been absolutely reliant on him. He had made, so to speak, both the inside and the limits of her universe. She was analyzed a clinical burdensome and experienced mental hospitalization. From that point forward, in the course of recent years, until the opportunity she went under the psychotherapeutic consideration of Dr. Banks, she had been hospitalized in any event on 10 additional events. During that time she had been under the watchfu l eye of in any event nine distinct psychotherapists, therapists, social laborers, and clinicians as an outpatient, one of whom were to mishandle her explicitly. Â

The Equity of Water Distribution Assignment Example | Topics and Well Written Essays - 1000 words

The Equity of Water Distribution - Assignment Example In 1998, Houston’s water clients expend 1.35 million cubic meters of water every day. The previous greatest every day pumpage, recorded in July of 1986, was 1.79 million cubic meters for every day. All the more as of late, be that as it may, Houston had siphoned a record-breaking record high of 1.96 million cubic meters for every day, during the warmth wave in 1998. Houston utilizes water for retail utilization, water system, and modern purposes. Directly, long droughts have required preservation endeavors to broaden accessible water gracefully. For the Sugar Land region, to diminish top interest and all out water utilization, Sugar Land inhabitants are approached to water yards as indicated by the accompanying calendar: Surat is situated at the mouth of the stream in floodplains and during the rainstorm season, it is the site of the convergence of water stream. Water is utilized for horticulture, drinking, water flexibly businesses and vitality age. Its water flexibly is given by the Ukai dam, which, put something aside for the yearly lean season, isn't shy of gracefully because of the yearly storm that brings substantial precipitation and flooding (Bhat, 2011). As of late the Maranoa Regional Council presented another water evaluating framework for private and business clients. The new utilization based framework requires the installment of a fixed assistance expense for progressing association, to which is included a charge for each kilolitre of water utilized. Be that as it may, the Council has barred Surat from the new evaluating framework; rather, the occupants and business will be distributed a yearly volume for use, with the overabundance charge to apply just for the utilization of extra kilolitres over the portion. The framework is intended to take care of the higher fixed expenses of working Surat’s double reticulation framework and stream water treatment process. The greater expense of treatment is because of the way that Surat’s water is drawn from the Balonne waterway rather than a drag, requiring a progressively costly technique for treatment.

DUI vs. DWI Whats the Difference

DUI vs. DWI What's the Difference Addiction Alcohol Use Drunk Driving Print Driving Under the Influence vs. Intoxicated By Buddy T facebook twitter Buddy T is an anonymous writer and founding member of the Online Al-Anon Outreach Committee with decades of experience writing about alcoholism. Learn about our editorial policy Buddy T Medically reviewed by Medically reviewed by Steven Gans, MD on February 09, 2017 Steven Gans, MD is board-certified in psychiatry and is an active supervisor, teacher, and mentor at Massachusetts General Hospital. Learn about our Medical Review Board Steven Gans, MD Updated on October 10, 2019 More in Addiction Alcohol Use Drunk Driving Binge Drinking Withdrawal and Relapse Children of Alcoholics Addictive Behaviors Drug Use Nicotine Use Coping and Recovery DUI is an acronym for driving under the influence. DWI stands for driving while intoxicated, or in some cases, driving while impaired. The terms can have different meanings or they can refer to the same offense, depending on the state in which you were pulled over.   In any case, DUI and DWI both mean that a driver is being charged with a serious offense that risked the health and safety of himself and others. They can apply not only to alcohol and recreational drugs  but also to driving when your prescription drugs impair your abilities. Its also important to understand that one is not worse than the other and that both can have a big effect on your life. Use of Terms DUI vs. DWI Differ From State to State Depending on state law, the two terms are both used to describe impaired or drunken driving. Some state laws refer to the offense of drunken driving as a DUI while others call it a DWI. It gets tricky when states use both terms. Quite often, they will refer one to alcohol and the other to impairment by drugs or an unknown substance and the meaning can flip-flop from state to state. In some states, DWI refers to driving while intoxicated of alcohol with a blood alcohol content (BAC) over the legal limit, while DUI is used when the driver is charged with being under the influence of alcohol or drugs.?? In other states where both terms are used, DWI means driving while impaired (by drugs, alcohol, or some unknown substance), while DUI means driving under the influence of alcohol. Its best to check the definitions of the state youre in. OUI and OWI There are other acronyms for drunk driving. OUI, or operating under the influence, is used in only three states: Maine, Massachusetts, and Rhode Island. OWI is an acronym for operating while intoxicated  which is used in some jurisdictions. The operating distinction encompasses more than just driving the vehicle. Even if the vehicle is stopped and not running, someone can be charged with operating under the influence. Blood-Alcohol Concentration Isnt the Only Factor in Determining Impaired Driving Any of these charges mean the arresting officer has reason to believe the driver is too impaired to continue to drive. In some jurisdictions, drivers can be charged with impaired driving (or driving under the influence) even if they do not meet the blood alcohol concentration levels for legal intoxication.?? For example, if you fail a field sobriety test or otherwise show signs of impairment, you can be charged with driving while impaired even if your blood-alcohol concentration is under the legal limit of 0.08. Drugged Driving Is Impaired Driving If you appear to be impaired by the arresting officer, but your breathalyzer test shows that you are not under the influence of alcohol, he may suspect that you have been using drugs and this is impairing your driving ability. These include prescription and nonprescription medications in addition to illegal drugs. The officer can call a Drug Recognition Expert (DRE) to the sceneâ€"or he may be one himselfâ€"to perform a series of tests.?? If the DRE officers multi-step evaluation process determines that you are indeed under the influence of drugs, you can be charged with DWI or DUI. The charge depends on what the state calls the offense of drugged driving. Taking prescription or nonprescription medications can impair your driving ability. You are at risk of drugged driving charges even when you have not had a sip of alcohol. Consequences of an Impaired Driving Arrest No matter what the offense is called in your jurisdiction, if you are arrested for impaired driving, you will be facing serious consequences. If you are convicted or plead guilty, you will probably lose your drivers license and pay fines and court fees. For a second offense, you may spend some time in jail. It is also likely that you will be placed on probation and be required to perform community service. To get your drivers license back, you will probably have to attend defensive driving classes. In most states, you will probably undergo an evaluation of your drinking or substance use patterns  as well. Based on the results of that evaluation, you may have to take part in a drug or  alcohol treatment program. That program could range from attending a few support group meetings like  Alcoholics Anonymous  to entering a residential treatment facility. The Ongoing Effects of a DUI or DWI Conviction When you get your drivers license back, you will likely need SR-22 insurance. This could double or triple your premiums, depending on the laws in your state. On average, you can expect to pay higher premiums for three years. Also, depending on the state in which you reside, you may be required to have an ignition interlock device installed on your vehicle. You will not be able to start your car unless you blow into the device and it determines you have not been drinking alcohol. This requires that you pay for the device, its installation, and a monthly monitoring fee.?? The bottom line is that getting arrested for driving under the influence is a time-consuming and very expensive ordeal. It is, however, 100 percent avoidable. Just dont get behind the wheel while you are drinking or taking any type of drug. This includes any prescription medications that warn about impaired driving or any that may affect your attention or focus, or cause drowsiness. A Word From Verywell You can protect your health and safetyâ€"as well as that of othersâ€"by  never driving after drinking any amount of alcohol. Your abilities will be impaired even if your blood alcohol content is below the legal limit.?? If you are taking any prescription or illicit drugs, its best not to get behind the wheel, either. The laws are in place to avoid potentially dangerous situations that are far worse than a DUI or DWI conviction.

The Importance Of A Teacher s Tools - 1371 Words

A Teacher’s Tools BACKGROUND The purpose of a teacher is to plan and organize a learning environment that guides students to achieve their academic potential. Teachers are required to have certain skills, knowledge, and tools. According to Dawn McKay states, â€Å"A teacher instructs students in subjects such as science, mathematics, language arts, social studies, art, and music, and then helps them apply those concepts. Teachers work in public or private elementary schools, middle schools, and high schools. Those working in middle and high schools usually specialize in teaching one subject.† Teachers teach students how to accomplish something, talk to students to convey information effectively, use learning strategies, active listening,†¦show more content†¦I walked into the classroom and saw other teachers carrying identification lanyard cards. Most of the teachers carried keys which I believe their keys to be able to enter their classroom and or building. I saw that the teacher carried a ring which I assume she is married. They also carried pens and notebook planners and talked about the schedule plan for the next week because a teacher was going to be gone so they needed to adjust certain things. Another teacher asked if they could borrow computers that students are able to use in their classrooms, so teachers are always sharing their tools. I sat down and looked around. The classroom was set for a small group with different type of stations. The teacher had a lecture desk and had a couple of papers there. It seemed like she was comfortable using it when lecturing. There were a large amount of reading books. In the teacher’s board was objectives for the students. In the teacher s desk there were many pictures of her family and of students’ drawings/notes. In her desk there were many papers and books stacked on top of each other and there were compartment files that had more papers and books. I saw projects all around the classroom that 7th graders and 8th graders completed. There were posters that encouraged students learning and thinking. There were 4 computer stations. While in classroom environment, students and teachers carried pencils, colored pencils,Show MoreRelatedCurriculum Based Learning, Data Informed Decisions And World Class Standards And Personalized Professional Development767 Words   |  4 Pagesdistrict administrators normally make these decisions for schools. World-Class Standards and Personalized Professional Development is defined as a new tool for teachers, as they are now conducting training by using computers instead of in-person. In today’s society teachers are taking the initiative of educating themselves professionally. Chandrasekaran, S., Al-Ameri, R. (2016). Assessing Team Learning Practices in Project/Design Based Learning Approach. International Journal of Engineering PedagogyRead MoreAnnotated Bibliography Ni Technology Education781 Words   |  4 Pagesdistrict administrators normally make these decisions for schools. World-Class Standards and Personalized Professional Development is defined as a new tool for teachers, as they are now conducting training by using computers instead of in-person. In today’s society teachers are taking the initiative of educating themselves professionally. Chandrasekaran, S., Al-Ameri, R. (2016). Assessing Team Learning Practices in Project/Design Based Learning Approach. International Journal of Engineering PedagogyRead MoreReading Comprehension Of English Language Learners At Harlem Success Academy1292 Words   |  6 PagesAs time Prevailed, school s, students and teacher’s expectations increased; we have noticed a decrease in academic success. Our schools have many different types of learners who are required to meet the same standard at the end of the year to be considered for promotion. In common classrooms, there are about twenty five- thirty students per one teacher. However, we must keep in mind that students learn differently and at a different pace. Unfortunately, there are standards and expectations studentsRead MoreReflection on the Integration of Technology in the Classroom Essay1255 Words   |  6 PagesThe following reflective essay will focus on technology and its importance in addressing the needs of digital learners. The essay will begin by addressing ways reasons for the integration of technology in education, as well as discussing ways in which teachers can use technology to enhance learning and student engagement. Secondly, the essay will examine how teachers can become part of the learning process by empowering students to serve as knowledge brokers. The essay will close by assessing waysRead MoreTeachers Can Start Using Technology833 Words   |  4 Pages1- Active: Teachers can start using technology not only to deliver a lesson but also let students to engage with the technological tools. For instance, using smart-board is becoming more popular day by day, and teachers are using it in their teaching activities. However, just showing videos on smart-boards to students relevant to the context that they are learning will not enhance students’ engagement and learning. Thus, teachers can allow students to deliberately engage with technological devicesRead MoreTaking a Look at Behaviourism635 Words   |  3 Pagesrecess time The teacher can reward a child, who is on toilet training, with a sticker or stamp to reinforce positive behaviour. What is Constructivism: learners construct knowledge for themselves Principles of constructivism: Learning is defined as the acquisition of new information which can be recalled later Role of teacher as a helper to construct knowledge by proving different tools Learner as an active participant rather than passive recipient of knowledge Gives importance to native languageRead MoreCreating A Data Driven Decision Making School Essay1339 Words   |  6 PagesLeadership, P.O Box 998 Normal, Alabama Contact: tcrook2@bulldogs.aamu.edu Abstract Collecting data has been a decades old practice of educators. The No Child Left Behind (NCLB) Act prompted front-runners in education to ascertain the importance of data in jump starting and carrying out school improvement plans. Student achievement across all socioeconomic frontiers in low performing schools was the basis for this legislation. Districts across the country were ushered into using data toRead MoreClassroom s : Run By Incentives896 Words   |  4 Pageswhile observing the first-grade classroom, the teacher was giving out points to groups. These points were given to students depending where they were sitting as a group. However, she will also remove points if one of the group s member was not following direction, which affected the entire group. At the end of the day, the group with most points will get a special treat. The treat consisted of either extra computer time, a candy, or a small toy. The teacher used incentives to keep her students on taskRead MoreAccountabi lity : High Performance And Not Fear Or Stress870 Words   |  4 Pagesindividually. â€Å"Accountability permeates education in the United States while focusing on process and product in education† (Thurlow, 2009). Responsibilities are shared with educational leaders, administrators, teachers, other school staff, and as well as students. While this term carries great importance, many educators may not be aware of its origin. According to the online periodical, Educational Research, written by Mintrop and Sunderman (2009), â€Å"The federal accountability system, made universal throughRead MoreHow Do Digital Media Affect The Classroom?1716 Words   |  7 Pageshow it works when it is in people s minds and mouths. In this essay I will firstly be explaining my understanding of the topic sentence, as well as discussing how it is important for a teacher to understand language, both as an object, as its parts as well as language as a whole, in motion. I will then be talking about how digital media in the classroom can provide a way to involve all children over coming the differences in language they may have, but that a teacher would have to keep in mind that

Violence at Work Westside Health Sytems - 1413 Words

Violence At Work: Westside Health Systems Located in Chicago, Illinois is a private nonprofit health care system called â€Å"Westside Health Systems†. The company consisted of a hospital, a nursing home and 5 minor emergency clinics. Maryanne Walker is the Director of Pharmacy Services, and oversees the main pharmacy located in the hospital, a pharmacy in the nursing home and four satellite pharmacies. She directly reports to her supervisor Nancy Smith. Maryanne is one of 5 total supervisors in Pharmacy Services. Rhonda Carter the inventory supervisor , is the direct supervisor of both employees involved in the allegation : Susan Miller and Brenda Lawson, both pharmacy technicians in the receiving area. The episode claimed that Susan had†¦show more content†¦Skills in taking disciplinary actions. Basic skills in handling crisis situations. Basic emergency procedures, including who to call and what support resources and services are available. Appropriate screening of pre-employment references. Basic skills in conflict resolution. 2. In light of this incident, should West-side change any of its standards of behavior policies or Corrective action policies? Explain. The only changes that I would suggest to West-Side regarding their standards of behavior policies would include: In any verbal counseling, both the employees direct supervisors should be present along with a member of upper management. This would ensure that no favoritism is being placed in one employee over another. Further, I would ensure that employees sign a statement acknowledging the fact that verbal counseling was undertaken. In written counseling, I again would require that each employee sign a statement that they received said written counseling. I am not sure how well written counseling would be as it does not require personal interaction between the employer and employee. Include a statement that indicates all parties involved will be required to meet management to discuss the matter. Failure to do so may require said employee to be reprimanded for simply refusing said meeting. The system of correction that West-side has is not a progressive one. I would suggest that it is progressive. If an employee is

Selecting Employee Free Essays

Learn how to select and hire the best employees for your open positions. Selection and evaluation techniques are explored that help you pick among qualified candidates. Employee selection processes are critical to hiring a superior staff. We will write a custom essay sample on Selecting Employee or any similar topic only for you Order Now Learn to improve your employee selection methods. 10 Tips for Hiring the Right Employee Top Ten Tips for Selecting and Hiring the Right Employee Hiring the right employee is a challenging process. Hiring the wrong employee is expensive, costly to your work environment, and time consuming. Hiring the right employee, on the other hand, pays you back in employee productivity, a successful employment relationship, and a positive impact on your total work environment. Hiring the right employee enhances your work culture and pays you back a thousand times over in high employee morale, positive forward thinking planning, and accomplishing challenging goals. This is not a comprehensive guide to hiring an employee. But, these are key steps to hiring the right employee. 1. Define the Job Before Hiring an Employee Hiring the right employee starts with a job analysis.  The job analysis enables you to collect information about the duties, responsibilities, necessary skills, outcomes, and work environment of a particular job. The information from the job analysis is fundamental to developing the job description for the new employee. The job description assists you to plan your recruiting strategy for hiring the right employee. Job HiringEasy Search Posting: AyosDito Free Job Posting, No Sign Ups! www. AyosDito. ph Interview Strategy GuideGet a Free interview strategy for HR professionals. hr. mcleanco. om/interview-guide Employment ScreeningInternational Background Checks No hidden fees or minimum orders 2. Plan Your Employee Recruiting Strategy With the job description in hand, set up a recruiting planning meeting that involves the key employees who are hiring the new employee. The hiring manager is crucial to the planning. At this meeting, your recruiting strategy is planned and the execution begins. Teams that have worked together frequently in hiring an employee can often complete this step via email. 3. Use a Checklist for Hiring an Employee  This checklist for hiring an employee will help you systematize your process for hiring an employee. Whether it’s your first employee or one of many employees you are hiring, this checklist for hiring an employee helps you keep track of your recruiting efforts. The checklist for hiring an employee keeps your recruiting efforts on track and communicates progress to interested employees and the hiring manager. 4. Recruit the Right Candidates When Hiring an Employee You can develop relationships with potential candidates long before you need them when hiring an employee.  These ideas will also help you in recruiting a large pool of candidates when you have a current position available. The more qualified candidates you can develop when hiring an employee, the more likely you are to locate a qualified potential employee. Read on to discover the best ways to develop your talent pool when hiring an employee. 5. Review Credentials and Applications Carefully The work of reviewing resumes, cover letters, job applications, and job application letters starts with a well-written job description.  Your bulletted list of the most desired characteristics of the most qualified candidate was developed as part of the recruiting planning process. Screen all applicants against this list of qualifications, skills, experience, and characteristics. You’ll be spending your time with your most qualified candidates when hiring an employee. And, that is a good use of your time. 6. Prescreen Your Candidates The most important reason to prescreen candidates when hiring an employee is to save the interviewing and selection committee time.  While a candidate may look good on paper, a prescreening interview will tell you if their qualifications are truly a fit with your job. Additionally, in a prescreening interview, you can determine whether their salary expectations are congruent with your job. A skilled telephone interviewer will also obtain evidence about whether the candidate may fit within your culture – or not. 7. Ask the Right Job Interview Questions The job interview is a powerful factor in hiring an employee. The job interview is a key tool employers utilize in hiring.  The job interview questions asked are critical in magnifying the power of the job interview to help you in hiring the right employee. Interview questions that help you separate desirable candidates from average candidates are fundamental when hiring an employee. Job interview questions matter to employers. Here are sample job interview questions. 8. Check Backgrounds and References When Hiring an Employee Effective background checks are one of the most important steps when hiring an employee. You need to verify that all the presented, sterling credentials, skills, and experience are actually possessed by your candidate.  The background checks must include work references, especially former supervisors, educational credentials, employment references and actual jobs held, and criminal history. Other background checks when hiring an employee, such as credit history, must be specifically related to the job for which you are hiring an employee. 9. Extend a Job Offer The job offer letter is provided to the candidate you have selected for the position. Most frequently, the candidate and the organization have verbally negotiated the conditions of hire and the job offer letter confirms the verbal agreements about salary and benefits.  The more senior the position, however, the more likely the job offer will turn into a protracted negotiation about salary, benefits, employment termination, bonus potential, severance pay, stock options, and more. 10. Use Effective Employment Letters These sample employment letters will assist you to reject job candidates, make job offers, welcome employees, and more when hiring an employee. Use these sample employment letters to develop the employment letters you use in your organization when hiring an employee. New Employee Orientation: Employee Onboarding New employee orientation is the process you use for welcoming a new employee nto your organization. New employee orientation, often spearheaded by a meeting with the Human Resources department, generally contains information about safety, the work environment, the new job description, benefits and eligibility, company culture, company history, the organization chart and anything else relevant to working in the new company. New employee orientation often includes an introduction to each department in the company and training on-the-job. New employee orientation frequently includes spending time doing the jobs in each department to understand the flow of the product or service through the organization. Tips for a Better New Employee Orientation When we orient new hourly (non-exempt) employees, we provide a standard HR couple of hours on policies, procedures, company history, goals, culture, punching in and work rules. We give a company tour and hourly employees then train and cross-train on the job. Managerial and salaried (exempt) employees participate in an orientation that is custom-designed for them. It includes the above information that is received by all employees. Additionally, their orientation may last one to two weeks and it enables them to meet the whole organization, their direct reports and more. They should leave this orientation with a clear picture of the organization, its challenges, its goals and their opportunity to assist with progress. It is challenging to make sure salaried employees have the chance to do the orientation while also beginning their new job. Neither can be put on hold. My current new director spent the morning helping to write an RFP for a potential customer rather than attending his scheduled meetings. This is okay, but I don’t want his orientation to get off track. It provides fundamental information he needs to succeed in this organization. From an HR perspective, this may not be ideal for making sure he gets the organization overview, but it is ideal for helping him integrate quickly into the working business of the company – and that’s the point. Right? The best orientation I have ever known was instituted at Edgewood Tool and Manufacturing. Every manager who hired a new employee was required to write a 120 day orientation for the new employee. It involved one action a day. Actions included meeting the Director of Quality, calling on a customer and having lunch with the CEO. You can bet that new employee was thoroughly welcomed and integrated into the organization after 120 different orientation events. Orientation and Training of New Employees New employee orientation effectively integrates the new employee into your organization and assists with retention, motivation, job satisfaction, and quickly enabling each individual to become contributing members of the work team. New Employee Welcome Letter-A welcome letter to a new employee who has accepted your job offer confirms the employe’s decision to accept the position. The welcome letter helps the new employee feel wanted and welcomed. Depending on the goal of your new employee welcome letter, these sample welcome letters will give you a template. See sample welcome letters for new employees. Onboarding-Onboarding is the process of acquiring, accommodating, assimilating and accelerating new team members, whether they come from outside or inside the organization. Effective onboarding of new team members is one of the most important contributions any hiring manager or HR professional can make to long-term success. Onboarding done right drives new employee productivity, accelerates results, and significantly improves talent retention. Yet few organizations manage the pieces of onboarding well. Purposes of Orientation Employers have to realize that orientation isn’t just a nice gesture put on by the organization. It serves as an important element of the recruitment and retention process. Some key purposes are: * To Reduce Startup Costs: Proper orientation can help the employee get â€Å"up to speed† much more quickly, thereby reducing the costs associated with learning the job. To Reduce Anxiety: Any employee, when put into a new, strange situation, will experience anxiety that can impede his or her ability to learn to do the job. Proper orientation helps to reduce anxiety that results from entering into an unknown situation, and helps provide guidelines for behavior and conduct, so the employee doesn’t have to experience the stress of guessing. * To Reduce Employee Turnover: Employee turnover increases as employees feel they are not valued, or are put in positions where they can’t possibly do their jobs. Orientation shows that the organization values the employee, and helps provide the tools necessary for succeeding in the job. * To Save Time for the Supervisor: Simply put, the better the initial orientation, the less likely supervisors and co-workers will have to spend time teaching the employee. * To Develop Realistic Job Expectations, Positive Attitudes and Job Satisfaction: It is important that employees learn as soon as possible what is expected of them, and what to expect from others, in addition to learning about the values and attitudes of the organization. While people can learn from experience, they will make many mistakes that are unnecessary and potentially damaging. The main reasons orientation programs fail: The program was not planned; the employee was unaware of the job requirements; the employee does not feel welcome. Employee orientation is important – orientation provides a lot of benefits, and you can use feedback to make your orientations even better. Use Training and Development to Motivate Staff Building Your Employee Training and Development Program Want to keep your staff motivated about learning new concepts? The quality and variety of the employee training you provide is key for motivation. Reasons for employee training range from new-hire training about your operation, to introducing a new concept to a workgroup to bringing in a new computer system. Whatever your reason for conducting an employee training session, you need to develop the employee training within the framework of a comprehensive, ongoing, and consistent employee training program. This quality employee training program is essential to keep your staff motivated about learning new concepts and your department profitable. Essential Components of Employee Training Programs A complete employee training program includes a formal new hire training program with an overview of the job expectations and performance skills needed to perform the job functions. A new hire training program provides a fundamental understanding of the position and how the position fits within the organizational structure. The more background knowledge the new associate has about how one workgroup interrelates with ancillary departments, the more the new associate will understand his or her impact on the organization. Another aspect of a comprehensive employee training program is continuing education. The most effective employee training programs make continuing education an ongoing responsibility of one person in the department. This is an important function that will keep all staff members current about policies, procedures and the technology used in the department. New Hire Training A solid new hire training program begins with the creation of an employee training manual, in either notebook format or online. This manual acts as a building block of practical and technical skills needed to prepare the new individual for his or her position. In order for the department to understand current policies and procedures, a manager must ensure the department manuals or online employee training are kept current. This includes any system enhancements and / or change in policy or procedure. In addition, keep the user in mind when designing training manuals or online training; keep the employee training material interesting for the learner. Use language that is not â€Å"corporate† and include images and multi-media. Much of this employee training and reference material belongs online these days in a company Intranet. But, if your organization is not ready to embrace the online world, keep the manuals up-to-date and interesting. When possible, in computer training, incorporate visual images of the computer screen (multi-media screen capture) to illustrate functions, examples, and how tos. On the Job Training Another form of new hire training includes having the new associate train directly next to an existing associate. Some call this On the Job Training (OJT) or side-by-side training. This type of employee training allows the new associate to see first hand the different facets of the position. Also, OJT allows the new hire the opportunity to develop a working relationship with an existing associate. This type of employee training reinforces concepts learned in the initial training and should be used to reinforce and apply those same learned concepts. Continuing Education in Employee Training A continuing education program for a department is just as important as the new hire training. When training a new associate, I have found that they will only retain approximately 40 percent of the information learned in the initial training session. Therefore, a continuous effort must be placed on reminding the staff about various procedures and concepts. This continuing education can be formal or informal. (The author’s preference is always with a more informal approach. ) The formal, or traditional approach, to employee training often includes a member of management sending a memo to each associate. The informal, and often more appealing approach to a visual learner, is to send a one-page information sheet to staff. This information sheet, called a training alert, should be informative and presented in a non-threatening manner. Therefore, if the policy or procedure changes, the informal approach would better prepare the department to receive this presentation. New Employee Training – Is It Worth The Investment Getting off on the right foot Many companies provide some sort of introductory training or orientation for most of their new employees. It may take the form of an older employee assigned to show the new employee â€Å"the ropes. † Or it may be left to the HR department or the individual’s new supervisor to show them where the coffee pot is and how to apply for time off. Many organizations, especially in government and academia, have created new employee training that is designed, exclusively or primarily, to provide mandated safety familiarization. Yet some companies in highly competitive industries recognize the value in New Employee Orientation (NEO) that goes much farther. They require several weeks or even months of training to familiarize every new employee with the company, its products, its culture and policies, even its competition. There is a measurable cost to that training, but is it worth it? Let’s look at some of the issues. Some Background Facts The technology in the workplace is changing very rapidly and companies that can’t keep up will drop out of competition. A survey by the Ontario (Canada) Skills Development Office found 63% of the respondents planned to â€Å"introduce new technology into the workplace that would require staff training. A third of the respondents included â€Å"improving employee job performance† and â€Å"keeping the best employees† as desired outcomes. The American Society for Training and Development (ASTD) reports that less than $1500 per employee was spent for training in 1996. The largest part of that (49 percent) was spent for technical and professional training. Only two percent was spent for New Employee Orientation and three percent on quality, competition and business practices training. How to cite Selecting Employee, Essay examples

Journeys free essay sample

The character that is seen to undergo the most profound change within the text ‘Away’ , by Michael Gow in my eyes, is Coral. We are introduced to Coral to be in an emotionally fragile condition, grieving the death of her son. She is seen to have alienated herself from society, and has a strained relationship with her husband Roy, unable to conform to his expectations. Coral’s psychological state is clearly depicted in the soliloquy Gow has utilised in Act One – Scene Three. Through her speech we understand that she is in an unstable state, as suggested at the beginning of the soliloquy, where she states, â€Å"When that woman woke up and saw that donkey at her feet I thought my heart would break. † This line generally depicts her detachment and alienation from society, through the inconceivable language used. Throughout irene gleesons life ( before the thought of the cinderella children project even started ) she had experienced pain and truma due to her past but she had a very strong faith in Christ which she believed helped her through her drakest days. We will write a custom essay sample on Journeys or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page With her fatherless home and harsh childhood, he grew to become a motherfigure to her family, even though later down the track her marriage broke down which resulting in her leaving her faith in god for a spiritual search, but through trialing other religions she returned to her Christian faith even more determined to help. She grew to know jesus as the only husband she needed. So before she knew it she had sold her dream beach house and her possessions to afford the big move with her caravan over to kitgum, Uganda. This is where the idea that gaining wisdom through out past experiences or journeys it may not seem relevant at the time but fundememntally help you when your struggling. The main character in Fight Club is the narrator and the main themes of the story are loneliness, materialism, and freedom from society. Tyler was created because of the lack of connection the narrator had with the people around him. The narrator was lonely and attended so many support groups because of it. He was not rejected at the support groups because the members thought he was sick just like they were. Materialism is a reoccurring theme as the narrator mentions how he has worked his entire life for the Ikea items in his apartment. He tried to fill the void in his life by buying worthless, meaningless stuff. People spend too much time working for things they do not need. The narrator comes to the conclusion that, â€Å"You are not your job or your possessions. † Only once a person realizes that can he or she finally let go and start living. â€Å"It’s only after you’ve lost everything,† Tyler says, â€Å"that you’re free to do anything. † In order to be free, we must not care about the stuff we own. When Tyler states â€Å"The things you own, end up owning you† it really opens the narrators eyes too see what he has based his life around †stuff†. Our whole lives are spent working to pay for stuff. If we did not have stuff to pay for, we would not have to work as hard and our time could be spent doing something more meaningful. This idea is also conveyed through the character Gwen from Michael Gow’s Away. A major conflict near the end of the story is between Tyler Durden and the narrator. The narrator discovered Tyler was a figment of his imagination and he wanted to stop him. The narrator wanted to get rid of Tyler, end Project Mayhem, and all of the Fight Clubs. Tyler did not want to leave and this conflict was resolved with the narrator shooting himself and killing Tyler. Tyler was created as someone the narrator always wanted to be. Tyler was the narrator’s hero and the narrator envied everything Tyler stood for. The narrator started to lose his own conformist identity and become more and more like Tyler. As the novel progressed, the narrator grew more miserable while becoming Tyler. Becoming Tyler was emotionally and physically draining for the narrator. Throughout the novel, the narrator is battling his former self and Tyler. He tried to find a happy medium between the two extremes. Towards the end, the narrator found this to be an impossible task as Tyler began to take over more and more. The narrator could not allow Tyler to continue controlling his life and destroying society so he had to shoot Tyler and himself in the process. â€Å"We need a break. We need a change† (Act Two – Scene Four). Gow has applied short and direct sentences to correspond Coral’s obstinacy and determination towards change. The repetition of ‘we need a†¦Ã¢â‚¬â„¢ reflects how Coral has prioritised change, and her views of change being a necessity. These techniques effectively suggest Coral’s acceptance towards change. Coral conjuncts the connotation of the holiday with positive implications, evident through her statement; ‘We’ll have a wonderful, wonderful time’ (Act Two – Scene Four). Repetition is prevalent once again in this excerpt, and has been used by Gow to portray her positive attitude in regards to the opportunity to change. This is where the idea that transformation on a journey comes from acceptance and letting go of past hurts that control you. The explicit meaning of the story in Fight Club is that Tyler made Fight Club for a way for men to overcome the frustrations of their professional and personal lives. There are no rules or limits as to how far Tyler will go to fulfil his goal. The implicit meaning is that Fight Club was made as an answer to the rejection of a consumer society is with the use of violence. Fighting was a way to free a man from society.

Questions on Platos Republic Essays

Questions on Platos Republic Essays Questions on Platos Republic Essay Questions on Platos Republic Essay Essay Topic: Questions The Republic Bearing in mind that a certain form of hierarchy has evolved into being in every sort of society known the man, the concept of the state is not something most people seem to criticize. The idea that a central body has control over a substantial amount of people because of its superiority in terms of education, intellect or experience, is however not enough to satisfy my questions. The problem with democracy is that everyone is given an equal vote, irrespective of whether or not they are a stakeholder or not, or have the intelligence or knowledge to make smart decisions. Within the status quo, we see democracy being upheld and held sacrosanct by almost everyone, but it too has its failures. If within a representative democracy the common man is assumed to be smart enough to elect the right candidate to rule the country, then the idea that the common man is not smart enough to make decisions for himself is completely ignored. If we are assumed to be of sound state of mind and with reasonable intelligence enough for us to select someone to rule us, then why are we not smart enough to know how to make decisions for ourselves, uphold peace, and do everything the state does on an individual level circumventing the entire process of the selection of the state? If the possibility of evil doers and criminals is acknowledged, and the idea of how the state and its legis lative and law enforcement bodies are the only ones capable enough to protect us from these individuals because these bodies are the only ones that can be trusted to make sound decisions comes into play, then the fact of the matter is it is that WE are the ones who have elected them in the first place. All of these statesmen have been at one point in time, a part of us. If the civilians are trusted to make sound decisions about their leaders, then I wonder why Donald Trump has won over Nevada, New Hampshire, and South Carolina. It is hence, extremely possible for democracy to resu

Different Versions of the Birth of Dionysus

Different Versions of the Birth of Dionysus In Greek mythology, there are often different and conflicting versions of mythological events. The story of the birth of Dionysus is no different, and Dionysus complicates matters by having different names. Here are two versions of the birth of Dionysus and one of the related birth of Zagreus: From a union between Persephone and Zeus in serpent form sprang the horned god Zagreus. Jealous Hera persuaded the Titans to attack the infant god as he looked into a mirror. Not only did they tear him to pieces, but the Titans ate him all but his heart which Athena rescued. From this organ, the rest of the god was resurrected. Semele is impregnated by drinking a preparation made from the heart of Dionysus who had been torn to pieces by the Titans. [Pseudo-Hyginus, Fabulae 167]Most familiar is the story of Semeles impregnation by Zeus but failure to live long enough to give birth to the child. To save the fetus, Zeus sewed him inside himself and gave birth through his leg when the time came.(ll. 940-942) And Semele, daughter of Cadmus was joined with him in love and bare him a splendid son, joyous Dionysus, a mortal woman an immortal son. And now they both are gods. Hesiod, Theogony (trans. Evelyn-White) Homeric Hymn1 to Dionysus ((LACUNA))(ll. 1-9) For some say, at Dracanum; and some, on windy Icarus; and some, in Naxos, O Heaven-born, Insewn; and others by the deep-eddying river Alpheus that pregnant Semele bare you to Zeus the thunder-lover. And others yet, lord, say you were born in Thebes; but all these lie. The Father of men and gods gave you birth remote from men and secretly from white-armed Hera. There is a certain Nysa, a mountain most high and richly grown with woods, far off in Phoenice, near the streams of Aegyptus.((LACUNA))(ll. 10-12) ...and men will lay up for her many offerings in her shrines. And as these things are three, so shall mortals ever sacrifice perfect hecatombs to you at your feasts each three years.(ll. 13-16) The Son of Cronos spoke and nodded with his dark brows. And the divine locks of the king flowed forward from his immortal head, and he made great Olympus reel. So spake wise Zeus and ordained it with a nod.(ll. 17-21) Be favourable, O Insewn, Inspirer of frenzied women! we singers sing of you as we begin and as we end a strain, and none forgetting you may call holy song to mind. And so, farewell, Dionysus, Insewn, with your mother Semele whom men call Thyone.Source: The Homeric Hymns I. To Dionysus [3.4.3] But Zeus loved Semele and bedded with her unknown to Hera. Now Zeus had agreed to do for her whatever she asked, and deceived by Hera she asked that he would come to her as he came when he was wooing Hera. Unable to refuse, Zeus came to her bridal chamber in a chariot, with lightning and thunderings, and launched a thunderbolt. But Semele expired of fright, and Zeus, snatching the sixth-month abortive child from the fire, sewed it in his thigh. On the death of Semele, the other daughters of Cadmus spread a report that Semele had bedded with a mortal man, and had falsely accused Zeus and that therefore she had been blasted by thunder. But at the proper time, Zeus undid the stitches and gave birth to Dionysus, and entrusted him to Hermes. And he conveyed him to Ino and Athamas, and persuaded them to rear him as a girl.- Apollodorus 3.4.3

Benefits of Going Green Essay Example | Topics and Well Written Essays - 3000 words

Benefits of Going Green - Essay Example At a global level, sustainable development has gained importance ever since the United Nations Framework Convention on Climate Change. Based on the UNFCC Article 3.4, sustainable development needs to be integrated with national development policies by considering the need for economic development to address climate change problems. In this convention, the focus has been more on reduction of greenhouse gas emission and their targets than on sustainable development. After this, much discussion has been done on the links between sustainable development and climate change and the inclusion of sustainable development concept in environmental policy. The sustainable development requirements approach to environmental policy started in developed nations first. Reports show that greenhouse gas emissions are going on rising with the ongoing economic reforms that aim at improving economic growth (Mc Kibbin, 2004). Based on economic theory, there are two main mitigation measures to reduce the greenhouse gas emission. One is a tradable permit system for emission rights and the other is taxing carbon emissions.... Studies have shown that the permit system for emission rights developing nations will result in a rise in costs in terms of sacrificing economic growth since most economic activities rely on energy here. The shift from cheap fossil fuels to expensive non-carbon energy will adversely affect economic growth and development (Prasad and Kochher, 2009). Recent estimates show that the cost for investment in reducing greenhouse gas emissions by 550 MtCO2 in the major energy emitting sectors cost S$25billion, which is equivalent to the amount needed for the development goals here(Prasad and Kochher,2009). The next possible option is carbon taxing. Taxing can be practically problematic since the taxes will be imposed on not only the emissions that are removed on the margin but on all emissions. Consequently, the income transfers from the firms to government will be very much larger than the costs of greenhouse gas emission abatements (McKibbin, 2004).Thus it can be shown that any form of abat ement measure will impose significant costs in terms of sacrificing economic growth and development in developing nations. This will not be a problem for developed nations, which have already achieved a high level of development in terms of all the indicators while in developing nations, where most people live below the poverty line, sacrificing development means a lot. Based on the different scandals that arose in the recent years like Enron scandal, it is argued that business ethics is essential which plays a major role in the functioning of companies (Broomhill, 2007).Studies show the beneficial effects of green policy depend on the expectations of stakeholders and the constraints placed by the restrictive legislations produced by the states(Broomhill,

Expose Article Example | Topics and Well Written Essays - 500 words

Expose - Article Example However, the electrician did nothing and said he would come to fix it later. Many residents believe that short circuit catalysed the occurrence of the fire. After the fire, the casualties were moved to another labour camp In AlShahaniya. Also, the employees were promised 200 riyals above their April salaries as compensation for the damage. Many volunteers have been appealing for food and clothe donations for the displaced men. Firstly, the article highlighted the troubling facts about the living conditions of the employees in the labour camps. Kovessy (2015) indicates that over eight men live in one room and there are no fire safety measures in the labour camps such as fire extinguishers and emergency exits despite the overcrowding of the rooms. Questions have been raised about Qatar’s commitment to safeguarding the welfare of their labour force despite the enormous wealth of the country. The evident negligence of the camp supervisors should have been questioned in courts since it was the major cause of the fire. The majority of the Sri Lankan employees are opting to go home despite the Sri Lankan embassy encouraging them to go back to work. They claim that they still don’t trust the safety standards of the labour camp. It is not understandable why the Sri Lankan government is not doing enough to cater for the rights of their nationals. A spokesman for the Sri Lankan embassy stated that no one was killed in the disaster; however, unconfirmed reports indicated that two Bangladesh nationals were killed in the fire. The statement exposed how the Sri Lankan Government is trying to cover up Qatar despite the fact that their nationals are suffering. Despite the fact that, the spokesperson was indicating the fire was evidence for the substandard housing conditions for the labourers, they agreed to settle for only 200 Riyal per employee. The displaced employees only escaped with the clothes on their bodies and 200 riyals could not possibly compensate for

Flow Cytometry for the Evaluation of Semen

Flow Cytometry for the Evaluation of Semen State of the Art in Sperm Assessment Using Flow Cytometry Abstract Flow cytometry is emerging as a substantial tool in the domain of modern andrology for the routine analysis of spermatozoa. Recent application of flow cytometry in the artificial insemination industry especially for pig is a new approach. Until very recent, analysis of semen samples was routinely performed by microscopical evaluation and manual techniques by laboratory operators; analysis is inclined due to comprehensive variability among observers, influencing its clinical validity. During last decade, to evaluate farm animal semen, variety of new flow cytometric techniques have been intercalated which made possible a wide spread evaluation of several sperm functionality and characteristics. Here in this paper, an initiative has been taken to explore numerous current flow cytometry developments pressing for andrological tests. After the invention of flow cytometry, sperm evaluation by traditional (microscopic) means became questioned and avoided due to the robust advantages of flow cytometry over the microscopic methods. By the recent development of diverse fluroscence probes, flow cytometry became capable of analyzing number of sperm characteristics like viability, capacitation, acrosomal integrity, membrane permeability, membrane integrity, mitochondrial status, DNA integrity, decondensation of DNA and differences between gametes based on sex. The application of flow cytometry to their detection allows increased numbers of spermatozoa to be assessed over a short time-period, provides the opportunity of working with small sample sizes, increases the repeatability of data obtained, removes the subjectivity of evaluation and allows simultaneous assessment of multiple fluorochromes. Thus, flow cytometry is a technique capable of generating significantly novel data and allows the design and execution of exper iments that are not yet possible with any other technique. Nowadays, semen evaluation using laboratory analyses is very meaningful to the artificial insemination industry to provide the most desired quality product to customers. Future development of flow cytometric techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. In this paper, the main semen attributes that can be analyzed with fluorochromes and adapted for use with a flow cytometer will be reviewed and the relationship of these tests to fertility will be discussed. Introduction Up to now, semen evaluation is considered as the most important laboratory test that has enabled us to identify and predict clear-cut cases of fertility (Jarow et al., 2002), infertility or even of potential sub-fertility (Rodrà ­guez-Martà ­nez, 2007). Determination of the potential fertility of semen sample and, in the long run, of the male from which it has been collected is the ultimate goal of semen evaluations in clinically healthy sires. Now a days, many methods for the estimate the possible fertilizing capacity of a semen sample and, or in the word, of the male (reviewed by Dziuk 1996; Rodrà ­guez-Martà ­nez et al. 1997a; Rodrà ­guez-Martà ­nez and Larsson 1998; Saacke et al. 1998; Larsson and Rodrà ­guez-Martà ­nez 2000; Rodrà ­guez- Martà ­nez 2000, 2003; Popwell and Flowers 2004; Graham and Mocà © 2005; Gillan et al. 2005) are existing. The methods routinely accustomed for evaluation of the quality of a semen sample involved an evaluation of general appearance, volume, pH, sperm concentration, viability, morphology and motility. Most of these evaluations are based on microscopic analyses that only measure relatively a few numbers of spermatozoa within a population. In most of the cases, these are time-consuming; results obtained are controversial and are not translatable. It should also be noted that such conventional techniques are apt to extreme inter-ejaculate variation, even when the laboratory methodology has been standardized. In the wake of this information, new opportunities have arisen for the development of methods for the diagnosis of male infertility, many of which have been shown to exhibit a prognostic value that eludes conventional semen profiling. Moreover, ejaculated spermatozoa are nowadays handled for use in assisted reproductive technologies, such as the artificial insemination of chilled, frozen-thawed or sexed semen, and IVF. During this long processes, number of steps like semen extension, fluorophore loading, ultrav iolet and laser illumination, high-speed sorting, cooling and cryopreservation are followed, which create a scope to impose different degrees of change in sperm functionality followed by suffer of damage to sperm membranes, organelles or the DNA content. Therefore, although several assays have been developed to monitor these sperm parameters, recently it is being claimed by many groups that buck of those so-called procedures are incomplete, time consuming and laborious. Flow cytometry in diverse technical applications proposes many advantages for the analysis of sperm quality. Flow cytometry is a method where multiple fluorescences and light scattering can be induced allowing single cell or particles illumination in suspension while they flow very rapidly through a sensing area. The increasing use over the past decade of flow cytometry in the leading laboratories in human and veterinary andrology has dramatically increased our knowledge of sperm function under physiological and biotechnological conditions. Flow cytometers is capable to acquire data from several subpopulations within a sample in a few minutes, making it perfect for assessing heterogenous populations in a semen sample. Flow cytometry was initially developed in the 1960s, after that flow cytometry is performing automated separation of cells based on the unique recognition of cellular patterns in a population feasible (Hulett et al., 1969). Likewise, cellular patterns can be recognized by utilizing such a separation approach, in each cells within a population (Baumgarth and Roederer, 2000; Herzenberg et al., 2006). The first notion of flow cytometry development was for medical and clinical applications such as haematology and oncology. Although still much research is going on these medical areas and account for the vast majority of publications on this robust technique, but during the past few years it is being used in a diverse areas, such as bioprocess monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Together with elevated use in many areas, recent advancement of flow cytometry increased its application in the reproductive biology especially for andrology. Although flow cytometry may overestimate the population of unlabelled cells (Petrunkina and Harrison, 2009), plethora of research from our group in pig (Pena et al., 2003, 2004, 2005; Spjuth et al., 2007; Fernando et al., 2003; Saravia et al.,2005, 2007,2009; De Ambrogi et al., 2006; ) bull (Bergquist et al., 2007; Nagy et al., 2004; Januskauskas et al., 2003; Bergqvist et al., 2007; Hallap et al., 20 05, 2006;) stallion ( Kavak et al., 2003; Morrell et al., 2008) indicate that newly developed fluorescent stains and techniques of flow cytometry has made possible a more widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, flow cytometry is the current technical solution for rapid, precisely reproducible assessment of sperm suspensions. In this review we have described potentiality and scope of flow cytometry for the evaluation of semen, and the way in which this technique can be used in clinical applications for andrology based on some of our previous experiences. Definition of flow cytometry The definition of a flow cytometer is ‘an instrument which measures the properties of cells in a flowing stream or ‘an instrument that can measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream. In other word, cytometry is a method which measure physical and chemical attributes of cells or other particles. Such a measurement is made when cells or other particles pass in single file through some sort of measuring apparatus in a stream of fluid. The data obtained can be used to understand and monitor biological processes and develop new methods and strategies for cell detection and quantification. Compared to other traditional analytical tools, where a single value for each attribute is obtained for the whole population, flow cytometry provides data for each and every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not only to detect a particular cell type but also to find different subpopulations according to their structural or physiological parameters. Flow cytometry is a technique for measuring components (cells) and the properties of individual cells in liquid suspension. In essence, suspended cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse across the detector, and fluorescence, absorbance, and light scattering are among the cell properties that can be detected. Flow sorting allows individual cells to be sorted on the basis of their measured properties, and one to three or more global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or more excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry presents objective and precise results (Bunthof et al., 2001; Shleeva et al., 2002), which help to overcome the problems with the manual methods described above. Function and types of flow cytometry A flow cytometer is made of three main systems, fluidics, optics and electronics. ItI It can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment came in to function when it was used for measuring their DNA content (Evenson et al., 1980) and its application for analyzing semen has been increased rapidly in last decade. Flow cytometry is now applied for the evaluation semen such as sperm viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity, DNA status and so on. Continuous innovation of new fluorescent stains and techniques facilitated the flow cytometric evaluation of spermatozoa. Flow cytometry allows the observation of physical characteristics, such as cell size, shape and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of experimental repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry also has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, meaning that more than one sperm attribute can be assessed simultaneously. This feature has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the acceptable range of normality (Larsson and Rodriguez-Martinez, 2000) and it is the general statistics that the more sperm parameters can be tested, the more accurate the fertility prediction becomes (Amman and Hammerstedt, 1993). There are two main types of flow cytometers-analysers and sorters are in use. Together with data collection on cells, sorters have the potentiality to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec manufacture a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid detection, characterization and enumeration of microorganisms. Luminex is developing technology for multiplexed analyte quantitation using a combination of microspheres, flow cytometry and high speed digital processing. Advantages of FC compared to other conventional techniques to explore sperm structure and function Use of authentic assays in the fertility clinic and artificial insemination industries increasing day by day. In this respect, use of flow cytometry might be an important attempt to resolve sustaining problem with so called commonly used manual method for the semen analysis. An additional source of laboratory variation is the low number of sperms analyzed with such techniques. It is worth mentinign here that so called method deal only with few hundred sperm. When we deal with such a few sperm population, there is a possibility that obtained result might not be statistically significant (Russel and Curtis, 1993). The methods which are frequently used are enable to determine sperm concentration (Jorgensen et al., 1997), motility or morphology only (Keel et al., 2002). Objectivity, cell number measured, speed of count and precision are the advantages of flow cytometry to conventional light microscopy techniques (Spano and Evenson, 1993). The technique now a days has been used to determi ne a number of factors including those of acrosome status, membrane integrity, mitochondrial function as well as multiparameter measurement in human (Garrido et al., 2002). Flow cytometry has the ability to analyze thousands of cells in few minutes. In our series of studies, we demonstrated the feasibility and reproducibility of an automated method to evaluate sperm cell type, count, and viability in human boar samples. In our hand, the precision of the flow cytometric analysis is satisfactory in a diverse species (boar, bull, stallion etc), and the observed errors were significantly better than those obtained from the so-called manual methods. Although there are diverse benefits of flow cytometer for the analysis of semen, feasibility of applying flow cytometry sometimes restricted to researcher due to the high outlay and difficulties of operation associated with the requirement of a skilled operator. Further, a flow cytometer is very large and cannot resist shocks associated with movement, and it also requires much space in the laboratory. Whatever may be the limitation, the development of more affordable ‘‘bench-top flow cytometers in recent time raised the potential essentialities to semen analysis. If the further application of flow cytometric analysis is considered further, it might be seen that it is growing popularities as a technique for assessing more than one sperm attribute, simultaneously. Compared to traditional microscopic techniques, flow cytometry analysis is allowing to give a far more simplified and objective method of semen analysis, especially in relation to fertilization with acrosome reactivity potential of spermatozoa (Uhler et al., 1993; Purvis et al., 1990; Carver-Ward et al., 1996). A large number of different techniques to estimate sperm concentration have been reported. In the mid-1990s a series of fixed-depth disposable slides were evaluated as rapid and effective pieces of equipment for the estimate of sperm concentration. Data from a number of preliminary studies proposed that, at least in the 20-mm-depth format, such chambers resulted in a noticeable underestimate of sperm concentration compared to the gold standard (improved Neubauer hemocytometer). According to the World Health Organization that ‘‘such chambers, whilst convenient in that they can be used without dilution of the specimen, might lead to inaccuracy (World Health Organization, 1999). Data from Tomlinson and colleagues indicate that two proprietary disposable slides (Microcell, Conception Technologies, San Diego, Calif; Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) can result in a lower concentrations of sperm compared to the hemocytometer method (Tomlinson et al., 2001) . In contrast, plenty of reports document unacceptable differences between different laboratories and even between different individuals, although fewer studies attempt to address these issues. So, what is wrong? Improvement of semen quality testing has been emphasizing in some reports (Jorgensen et al., 1997; Keel et al., 2000). But due to low number of sperm evaluation by the conventional method results in poor reproducibility. These problems might be overcome when using flow cytometry. The validation of method is a challenge due to its essentiality of having specific, precise, objective, and accurate evaluation to establish a correlation of fertility data or to predict potential of a semen sample accurately (Amann, 1989). In a fertility clinic, precision of data in important as the result of semen analysis is frequently used to manage fertility of a patient and treatment of the unfertile couples. Thus, it is important to take into consideration within and between laboratory variations for successful infertility treatments. Sometimes its a matter of argument that compared to flow cytometry, fluorescent microscopy evaluate â€Å"patterns of fluorescence rather than the fluorescence intensity. Flow cytometer has the lack of ability to discriminate sperm containing a fluorescent marker bound to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993; Mortimer and Camenzind, 1989; Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with various lectins and indicated that there is almost no difference between methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosomal region of the sperm (Purvis et al., 1990; Holden and Trounson, 1991) and that other binding sites can be easily distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm. Additionally, conventional light microscopic semen assessment is increasingly being replaced by fluorescent staining techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (PenËÅ"a et al., 2001; Verstegen et al., 2002). Additional advantages over existing techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of somatic cells in a semen sample. Application of flow cytometry for sperm count Sperm count is an important predominant factor for the evaluation of sperm fertility potential. Accurate determination of sperm cell concentration is critical especially in AI industry because it provides assurance to customers that straws of extended semen contain the sperm numbers indicated which will help to decide appropriate doze especially for pig. Accuracy of sperm count is a common problem in the andrological laboratories and accurate measure of sperm concentration is particularly important for export in which verification of numbers may be required. Routine sperm counts can help to identify possible processing errors within a specific batch of semen or on a particular day, should those errors occur. As sperm counting procedures become more refined, routine counting can be used to monitor subtle changes in daily semen processing that might affect the number of sperm packaged in a straw. Every time new and more accurate methods for the sperm count determinations are coming and being replaced by the older ones. Some laboratories are trying the Maklerm counting chamber (Se if- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA); however, these techniques will likely have standard lems similar to those associated with the standard hemacytometers. Although hemacytometers are routinely used for sperm counts, due to the slow process and need for multiple measurements of each sample, the chance of error increase. Freund and Carol (ref) stated that a difference of 20% were not unusual between the determinations by the same technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicians. However, those instruments tend to include in the sperm count any somatic cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (Ref). Spectrophotometer is recently being used in the AI industries to assess sperm concentration by determining turbidity of a semen s ample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (Ref). The accuracy of this method depends on the methods used for spectrophotometer calibration. Although, sperm concentration can also be determined by spectrophotometrically, the debris present in the raw semen crease problem with misestimation. Sperm number in the frozen thawed semen is difficult to ascertain as most of the extender contain egg yolk particles, fats and other particles which affect measurement of sperm with electric cell counter or spectrophotometers (Evenson et al., 1993). On the other hand flow cytometry created possibilities of a rapid determination of sperm number in a precise form. It is the flow cytometry which can reduce intra-laboratory and inter-laboratory variation and conflict regarding sperm concentration assessment. Computer assisted semen analyzer is robust technique for analyzing sperm movement which can count sperm as well; but such an a nalyzer most of the cases use some counting chamber or hemacytometer which itself can generate error. Although, hemacytometer was originally developed for blood cell counting, its use is now diverse including andrological laboratories for sperm counting. Around two-decade ago flow cytometry was suggested for sperm numbers in straws of cryopreserved bull semen. Christensen et al. (-) observed similar results for sperm count with flow cytometry and hemocytometer for a number of species. Now a day a simultaneous determination of sperm viability and sperm concentration is possible which can avoid the chance of occurring differences between ejaculates leading lack of coordination with field fertility and laboratory analyses. Thus the present technology is more precise which can get rid of variation from handling the sperm sample and variation from pipetting and the analysis itself. Further, Prathalingam et al. (2006) concluded that there is similarities for sperm count result between flow cytometry and two newly approached method (image analysis and fluorescent plate reader) for sperm counting. Though, use of fluorescent plate was emphasized due to low cost and allowing large number of cells counting from a large number of ejaculates. Although flow cytometry has become a valuable instrument for andrological determinations, it is also blamed that sperm concentration by flow cytometry signify a higher value than the real one. The possibility arise might be due to that semen samples often contain some alien materials such as immature germ cells, epithelial cells, blood cells, cytoplasmic droplet, cellular debris etc. In the same way, frozen semen has higher chance to introduce such material as they contain diluents components especially egg yolk particles. These particles and cell debris might have frontal and side light scatter parameters those are similar to spermatozoa. Such sperm-count-overestimation problem arisen in our cases also, especially when we deal with frozen semen. Further it is also claimed that flow cytometry has a tendency to overestimate viable spermatozoa. We are also experienced with such trouble which we guess might be due to that egg particles of extender are considered as viable cell as for it s staining pattern. Our yet to publish data indicate that this problem can be mimic by a centrifugation process and by using low concentration sample for evaluation with flow cytometry. Very recently Petrunkina and Harrison (2009) proposed a mathematical equation for fixing this flow cytometric sperm counting. Thus much research is going on and we hope such discrepancy will completely be resolved near future to get advantage from this robust technology for sperm counting. Flow cytometry for detecting sperm intactness -Viability of spermatozoa The viability of spermatozoa is a key determinant of sperm quality and prerequisite for successful fertilization. Viability of spermatozoa can be assessed by numerous methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow cytometry estimate similar trend of viability. Current flow cytometric procedures are able to simultaneously evaluate sperm cell viability together with some other attributes. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994; Thomas et al., 1998), boars (Rodrà ­guez-Martà ­nez, 2007; Garner and Johnson, 1995; Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995; Songsasen et al., 199 7), poultry and wildfowl (Donoghue et al., 1995; Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999; Collins, 2000) and in fish (Martin Flajshans et al., 2004). Considerable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very bright in living sperm. Good agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test; the flow cytometric method showed a precision level higher than that of the manual method. One of the first attempts to assess sperm viability utilized rhodamine 123 for determining potentiality of mitochondrial membrane while ethidium bromide for membrane integrity through flow cytometry (Garner et al., 1986). Other combinations that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988; Watson et al., 1992); carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993; Thomas and Garner, 1994); and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990). The most generally used sperm viability stain combinations is SYBR-14 and PI at present. This stains are now sold commercially as live/dead viability kit. When these two stains are combinely used, the nuclei of viable sperm take fluoresce green and membrane integrity lost cells take red stain. This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995; Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the Live/Dead stain may similarly have no adverse affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been introduced to evaluate sperm concentration [11] and viability. Due to the small size and low cost, this instrument has been attracted for field measurements of both concentration and viability. In our hand this instrument was also became useful for the quick measurement of sperm concentration an d viability in stallion (Morrell et al., 2010). Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that operates in the ultraviolet light range, are less commonly used as this is not a standard feature on the smaller analytical machines. However, one alternative is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect nonviable bull spermatozoa by fluorometry and obtained an inverse correlation between the damaged cells per cent and the field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with motility. Fluorochromes used to assess sperm viability by both approach could be utilized in combination with each other. In that case, when CFDA is used combined with PI, three populations of cells as live, which are green; dead, which are red; and a third population which is stained with both and represents dying spermatozoa can be identified. This combination was found useful by Almlid and Johnson (1988) for frozen-thawed boar spermatozoa for monitoring membrane damage at the time of evaluation of various freezing protocols. Further, Harrison and Vickers (1990) also noticed that this combination with a fluorescent microscope is effective indicator of viability of fresh, incubated or cold-shocked spermatozoa in boar and ram. Contrasting to these, Garner et al. (1986) was failed to find a relationship between bull sperm viability and fertility when using combination of CFDA/PI . Flow cytometry for evaluating sperm viability appears to be a precious tool in the AI industry. When a high number of sperm is packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect. However, sperm viability might be more important when combined with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combined assessment of sperm viability and concentration appears to be useful in the wake of improving quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively bright fluorescence was found also in the mitochondrial sheath of living sperm. But the mechanism and mode of action by which SYBR-14 binds to the DNA of sperm is not known. It is know that PI stains nucleic acids by inte rcalating between the base pairs (Krishan, 1975). Viability stains can also be used in conjugation with fluorescently labeled plant lectins for simultaneous assessment of the plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). It is conceivable that assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garner et al. (5) found that insemination of boar sperm stained with SYBR-14 did not compromise fertilization or even the development of flushed porcine embryos in vitro. Non-viable sperms can be detected using the membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with damaged membranes. Plasma membrane which is intact will not permit these stains entering into the spermatozoa and staining the nucleus. Most frequently used stains include phenanthridines, for example propidium iodide (PI; (Matyus, 1984) ethidium homodimer-1 (EthD-1; (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). After a series of comparison between fertility of cryopreserved stallion spermatozoa with a number of laboratory assessments of semen quality as assessed by flow cytometry using PI, Wilhelm et al. (1996) concluded that viability is the single laboratory assay that correlated with fertility. -Sperm plasma membrane integrity Although the sperm plasma membrane covers the entire cell, it consists of three distinct membrane compartments, one which covers the outer acrosomal membrane, one which covers the post acrosomal portion of the sperm head, and one which covers the middle and principal pieces. Sperm membrane is directly or ind Flow Cytometry for the Evaluation of Semen Flow Cytometry for the Evaluation of Semen State of the Art in Sperm Assessment Using Flow Cytometry Abstract Flow cytometry is emerging as a substantial tool in the domain of modern andrology for the routine analysis of spermatozoa. Recent application of flow cytometry in the artificial insemination industry especially for pig is a new approach. Until very recent, analysis of semen samples was routinely performed by microscopical evaluation and manual techniques by laboratory operators; analysis is inclined due to comprehensive variability among observers, influencing its clinical validity. During last decade, to evaluate farm animal semen, variety of new flow cytometric techniques have been intercalated which made possible a wide spread evaluation of several sperm functionality and characteristics. Here in this paper, an initiative has been taken to explore numerous current flow cytometry developments pressing for andrological tests. After the invention of flow cytometry, sperm evaluation by traditional (microscopic) means became questioned and avoided due to the robust advantages of flow cytometry over the microscopic methods. By the recent development of diverse fluroscence probes, flow cytometry became capable of analyzing number of sperm characteristics like viability, capacitation, acrosomal integrity, membrane permeability, membrane integrity, mitochondrial status, DNA integrity, decondensation of DNA and differences between gametes based on sex. The application of flow cytometry to their detection allows increased numbers of spermatozoa to be assessed over a short time-period, provides the opportunity of working with small sample sizes, increases the repeatability of data obtained, removes the subjectivity of evaluation and allows simultaneous assessment of multiple fluorochromes. Thus, flow cytometry is a technique capable of generating significantly novel data and allows the design and execution of exper iments that are not yet possible with any other technique. Nowadays, semen evaluation using laboratory analyses is very meaningful to the artificial insemination industry to provide the most desired quality product to customers. Future development of flow cytometric techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. In this paper, the main semen attributes that can be analyzed with fluorochromes and adapted for use with a flow cytometer will be reviewed and the relationship of these tests to fertility will be discussed. Introduction Up to now, semen evaluation is considered as the most important laboratory test that has enabled us to identify and predict clear-cut cases of fertility (Jarow et al., 2002), infertility or even of potential sub-fertility (Rodrà ­guez-Martà ­nez, 2007). Determination of the potential fertility of semen sample and, in the long run, of the male from which it has been collected is the ultimate goal of semen evaluations in clinically healthy sires. Now a days, many methods for the estimate the possible fertilizing capacity of a semen sample and, or in the word, of the male (reviewed by Dziuk 1996; Rodrà ­guez-Martà ­nez et al. 1997a; Rodrà ­guez-Martà ­nez and Larsson 1998; Saacke et al. 1998; Larsson and Rodrà ­guez-Martà ­nez 2000; Rodrà ­guez- Martà ­nez 2000, 2003; Popwell and Flowers 2004; Graham and Mocà © 2005; Gillan et al. 2005) are existing. The methods routinely accustomed for evaluation of the quality of a semen sample involved an evaluation of general appearance, volume, pH, sperm concentration, viability, morphology and motility. Most of these evaluations are based on microscopic analyses that only measure relatively a few numbers of spermatozoa within a population. In most of the cases, these are time-consuming; results obtained are controversial and are not translatable. It should also be noted that such conventional techniques are apt to extreme inter-ejaculate variation, even when the laboratory methodology has been standardized. In the wake of this information, new opportunities have arisen for the development of methods for the diagnosis of male infertility, many of which have been shown to exhibit a prognostic value that eludes conventional semen profiling. Moreover, ejaculated spermatozoa are nowadays handled for use in assisted reproductive technologies, such as the artificial insemination of chilled, frozen-thawed or sexed semen, and IVF. During this long processes, number of steps like semen extension, fluorophore loading, ultrav iolet and laser illumination, high-speed sorting, cooling and cryopreservation are followed, which create a scope to impose different degrees of change in sperm functionality followed by suffer of damage to sperm membranes, organelles or the DNA content. Therefore, although several assays have been developed to monitor these sperm parameters, recently it is being claimed by many groups that buck of those so-called procedures are incomplete, time consuming and laborious. Flow cytometry in diverse technical applications proposes many advantages for the analysis of sperm quality. Flow cytometry is a method where multiple fluorescences and light scattering can be induced allowing single cell or particles illumination in suspension while they flow very rapidly through a sensing area. The increasing use over the past decade of flow cytometry in the leading laboratories in human and veterinary andrology has dramatically increased our knowledge of sperm function under physiological and biotechnological conditions. Flow cytometers is capable to acquire data from several subpopulations within a sample in a few minutes, making it perfect for assessing heterogenous populations in a semen sample. Flow cytometry was initially developed in the 1960s, after that flow cytometry is performing automated separation of cells based on the unique recognition of cellular patterns in a population feasible (Hulett et al., 1969). Likewise, cellular patterns can be recognized by utilizing such a separation approach, in each cells within a population (Baumgarth and Roederer, 2000; Herzenberg et al., 2006). The first notion of flow cytometry development was for medical and clinical applications such as haematology and oncology. Although still much research is going on these medical areas and account for the vast majority of publications on this robust technique, but during the past few years it is being used in a diverse areas, such as bioprocess monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Together with elevated use in many areas, recent advancement of flow cytometry increased its application in the reproductive biology especially for andrology. Although flow cytometry may overestimate the population of unlabelled cells (Petrunkina and Harrison, 2009), plethora of research from our group in pig (Pena et al., 2003, 2004, 2005; Spjuth et al., 2007; Fernando et al., 2003; Saravia et al.,2005, 2007,2009; De Ambrogi et al., 2006; ) bull (Bergquist et al., 2007; Nagy et al., 2004; Januskauskas et al., 2003; Bergqvist et al., 2007; Hallap et al., 20 05, 2006;) stallion ( Kavak et al., 2003; Morrell et al., 2008) indicate that newly developed fluorescent stains and techniques of flow cytometry has made possible a more widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, flow cytometry is the current technical solution for rapid, precisely reproducible assessment of sperm suspensions. In this review we have described potentiality and scope of flow cytometry for the evaluation of semen, and the way in which this technique can be used in clinical applications for andrology based on some of our previous experiences. Definition of flow cytometry The definition of a flow cytometer is ‘an instrument which measures the properties of cells in a flowing stream or ‘an instrument that can measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream. In other word, cytometry is a method which measure physical and chemical attributes of cells or other particles. Such a measurement is made when cells or other particles pass in single file through some sort of measuring apparatus in a stream of fluid. The data obtained can be used to understand and monitor biological processes and develop new methods and strategies for cell detection and quantification. Compared to other traditional analytical tools, where a single value for each attribute is obtained for the whole population, flow cytometry provides data for each and every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not only to detect a particular cell type but also to find different subpopulations according to their structural or physiological parameters. Flow cytometry is a technique for measuring components (cells) and the properties of individual cells in liquid suspension. In essence, suspended cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse across the detector, and fluorescence, absorbance, and light scattering are among the cell properties that can be detected. Flow sorting allows individual cells to be sorted on the basis of their measured properties, and one to three or more global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or more excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry presents objective and precise results (Bunthof et al., 2001; Shleeva et al., 2002), which help to overcome the problems with the manual methods described above. Function and types of flow cytometry A flow cytometer is made of three main systems, fluidics, optics and electronics. ItI It can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment came in to function when it was used for measuring their DNA content (Evenson et al., 1980) and its application for analyzing semen has been increased rapidly in last decade. Flow cytometry is now applied for the evaluation semen such as sperm viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity, DNA status and so on. Continuous innovation of new fluorescent stains and techniques facilitated the flow cytometric evaluation of spermatozoa. Flow cytometry allows the observation of physical characteristics, such as cell size, shape and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of experimental repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry also has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, meaning that more than one sperm attribute can be assessed simultaneously. This feature has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the acceptable range of normality (Larsson and Rodriguez-Martinez, 2000) and it is the general statistics that the more sperm parameters can be tested, the more accurate the fertility prediction becomes (Amman and Hammerstedt, 1993). There are two main types of flow cytometers-analysers and sorters are in use. Together with data collection on cells, sorters have the potentiality to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec manufacture a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid detection, characterization and enumeration of microorganisms. Luminex is developing technology for multiplexed analyte quantitation using a combination of microspheres, flow cytometry and high speed digital processing. Advantages of FC compared to other conventional techniques to explore sperm structure and function Use of authentic assays in the fertility clinic and artificial insemination industries increasing day by day. In this respect, use of flow cytometry might be an important attempt to resolve sustaining problem with so called commonly used manual method for the semen analysis. An additional source of laboratory variation is the low number of sperms analyzed with such techniques. It is worth mentinign here that so called method deal only with few hundred sperm. When we deal with such a few sperm population, there is a possibility that obtained result might not be statistically significant (Russel and Curtis, 1993). The methods which are frequently used are enable to determine sperm concentration (Jorgensen et al., 1997), motility or morphology only (Keel et al., 2002). Objectivity, cell number measured, speed of count and precision are the advantages of flow cytometry to conventional light microscopy techniques (Spano and Evenson, 1993). The technique now a days has been used to determi ne a number of factors including those of acrosome status, membrane integrity, mitochondrial function as well as multiparameter measurement in human (Garrido et al., 2002). Flow cytometry has the ability to analyze thousands of cells in few minutes. In our series of studies, we demonstrated the feasibility and reproducibility of an automated method to evaluate sperm cell type, count, and viability in human boar samples. In our hand, the precision of the flow cytometric analysis is satisfactory in a diverse species (boar, bull, stallion etc), and the observed errors were significantly better than those obtained from the so-called manual methods. Although there are diverse benefits of flow cytometer for the analysis of semen, feasibility of applying flow cytometry sometimes restricted to researcher due to the high outlay and difficulties of operation associated with the requirement of a skilled operator. Further, a flow cytometer is very large and cannot resist shocks associated with movement, and it also requires much space in the laboratory. Whatever may be the limitation, the development of more affordable ‘‘bench-top flow cytometers in recent time raised the potential essentialities to semen analysis. If the further application of flow cytometric analysis is considered further, it might be seen that it is growing popularities as a technique for assessing more than one sperm attribute, simultaneously. Compared to traditional microscopic techniques, flow cytometry analysis is allowing to give a far more simplified and objective method of semen analysis, especially in relation to fertilization with acrosome reactivity potential of spermatozoa (Uhler et al., 1993; Purvis et al., 1990; Carver-Ward et al., 1996). A large number of different techniques to estimate sperm concentration have been reported. In the mid-1990s a series of fixed-depth disposable slides were evaluated as rapid and effective pieces of equipment for the estimate of sperm concentration. Data from a number of preliminary studies proposed that, at least in the 20-mm-depth format, such chambers resulted in a noticeable underestimate of sperm concentration compared to the gold standard (improved Neubauer hemocytometer). According to the World Health Organization that ‘‘such chambers, whilst convenient in that they can be used without dilution of the specimen, might lead to inaccuracy (World Health Organization, 1999). Data from Tomlinson and colleagues indicate that two proprietary disposable slides (Microcell, Conception Technologies, San Diego, Calif; Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) can result in a lower concentrations of sperm compared to the hemocytometer method (Tomlinson et al., 2001) . In contrast, plenty of reports document unacceptable differences between different laboratories and even between different individuals, although fewer studies attempt to address these issues. So, what is wrong? Improvement of semen quality testing has been emphasizing in some reports (Jorgensen et al., 1997; Keel et al., 2000). But due to low number of sperm evaluation by the conventional method results in poor reproducibility. These problems might be overcome when using flow cytometry. The validation of method is a challenge due to its essentiality of having specific, precise, objective, and accurate evaluation to establish a correlation of fertility data or to predict potential of a semen sample accurately (Amann, 1989). In a fertility clinic, precision of data in important as the result of semen analysis is frequently used to manage fertility of a patient and treatment of the unfertile couples. Thus, it is important to take into consideration within and between laboratory variations for successful infertility treatments. Sometimes its a matter of argument that compared to flow cytometry, fluorescent microscopy evaluate â€Å"patterns of fluorescence rather than the fluorescence intensity. Flow cytometer has the lack of ability to discriminate sperm containing a fluorescent marker bound to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993; Mortimer and Camenzind, 1989; Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with various lectins and indicated that there is almost no difference between methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosomal region of the sperm (Purvis et al., 1990; Holden and Trounson, 1991) and that other binding sites can be easily distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm. Additionally, conventional light microscopic semen assessment is increasingly being replaced by fluorescent staining techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (PenËÅ"a et al., 2001; Verstegen et al., 2002). Additional advantages over existing techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of somatic cells in a semen sample. Application of flow cytometry for sperm count Sperm count is an important predominant factor for the evaluation of sperm fertility potential. Accurate determination of sperm cell concentration is critical especially in AI industry because it provides assurance to customers that straws of extended semen contain the sperm numbers indicated which will help to decide appropriate doze especially for pig. Accuracy of sperm count is a common problem in the andrological laboratories and accurate measure of sperm concentration is particularly important for export in which verification of numbers may be required. Routine sperm counts can help to identify possible processing errors within a specific batch of semen or on a particular day, should those errors occur. As sperm counting procedures become more refined, routine counting can be used to monitor subtle changes in daily semen processing that might affect the number of sperm packaged in a straw. Every time new and more accurate methods for the sperm count determinations are coming and being replaced by the older ones. Some laboratories are trying the Maklerm counting chamber (Se if- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA); however, these techniques will likely have standard lems similar to those associated with the standard hemacytometers. Although hemacytometers are routinely used for sperm counts, due to the slow process and need for multiple measurements of each sample, the chance of error increase. Freund and Carol (ref) stated that a difference of 20% were not unusual between the determinations by the same technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicians. However, those instruments tend to include in the sperm count any somatic cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (Ref). Spectrophotometer is recently being used in the AI industries to assess sperm concentration by determining turbidity of a semen s ample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (Ref). The accuracy of this method depends on the methods used for spectrophotometer calibration. Although, sperm concentration can also be determined by spectrophotometrically, the debris present in the raw semen crease problem with misestimation. Sperm number in the frozen thawed semen is difficult to ascertain as most of the extender contain egg yolk particles, fats and other particles which affect measurement of sperm with electric cell counter or spectrophotometers (Evenson et al., 1993). On the other hand flow cytometry created possibilities of a rapid determination of sperm number in a precise form. It is the flow cytometry which can reduce intra-laboratory and inter-laboratory variation and conflict regarding sperm concentration assessment. Computer assisted semen analyzer is robust technique for analyzing sperm movement which can count sperm as well; but such an a nalyzer most of the cases use some counting chamber or hemacytometer which itself can generate error. Although, hemacytometer was originally developed for blood cell counting, its use is now diverse including andrological laboratories for sperm counting. Around two-decade ago flow cytometry was suggested for sperm numbers in straws of cryopreserved bull semen. Christensen et al. (-) observed similar results for sperm count with flow cytometry and hemocytometer for a number of species. Now a day a simultaneous determination of sperm viability and sperm concentration is possible which can avoid the chance of occurring differences between ejaculates leading lack of coordination with field fertility and laboratory analyses. Thus the present technology is more precise which can get rid of variation from handling the sperm sample and variation from pipetting and the analysis itself. Further, Prathalingam et al. (2006) concluded that there is similarities for sperm count result between flow cytometry and two newly approached method (image analysis and fluorescent plate reader) for sperm counting. Though, use of fluorescent plate was emphasized due to low cost and allowing large number of cells counting from a large number of ejaculates. Although flow cytometry has become a valuable instrument for andrological determinations, it is also blamed that sperm concentration by flow cytometry signify a higher value than the real one. The possibility arise might be due to that semen samples often contain some alien materials such as immature germ cells, epithelial cells, blood cells, cytoplasmic droplet, cellular debris etc. In the same way, frozen semen has higher chance to introduce such material as they contain diluents components especially egg yolk particles. These particles and cell debris might have frontal and side light scatter parameters those are similar to spermatozoa. Such sperm-count-overestimation problem arisen in our cases also, especially when we deal with frozen semen. Further it is also claimed that flow cytometry has a tendency to overestimate viable spermatozoa. We are also experienced with such trouble which we guess might be due to that egg particles of extender are considered as viable cell as for it s staining pattern. Our yet to publish data indicate that this problem can be mimic by a centrifugation process and by using low concentration sample for evaluation with flow cytometry. Very recently Petrunkina and Harrison (2009) proposed a mathematical equation for fixing this flow cytometric sperm counting. Thus much research is going on and we hope such discrepancy will completely be resolved near future to get advantage from this robust technology for sperm counting. Flow cytometry for detecting sperm intactness -Viability of spermatozoa The viability of spermatozoa is a key determinant of sperm quality and prerequisite for successful fertilization. Viability of spermatozoa can be assessed by numerous methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow cytometry estimate similar trend of viability. Current flow cytometric procedures are able to simultaneously evaluate sperm cell viability together with some other attributes. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994; Thomas et al., 1998), boars (Rodrà ­guez-Martà ­nez, 2007; Garner and Johnson, 1995; Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995; Songsasen et al., 199 7), poultry and wildfowl (Donoghue et al., 1995; Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999; Collins, 2000) and in fish (Martin Flajshans et al., 2004). Considerable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very bright in living sperm. Good agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test; the flow cytometric method showed a precision level higher than that of the manual method. One of the first attempts to assess sperm viability utilized rhodamine 123 for determining potentiality of mitochondrial membrane while ethidium bromide for membrane integrity through flow cytometry (Garner et al., 1986). Other combinations that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988; Watson et al., 1992); carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993; Thomas and Garner, 1994); and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990). The most generally used sperm viability stain combinations is SYBR-14 and PI at present. This stains are now sold commercially as live/dead viability kit. When these two stains are combinely used, the nuclei of viable sperm take fluoresce green and membrane integrity lost cells take red stain. This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995; Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the Live/Dead stain may similarly have no adverse affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been introduced to evaluate sperm concentration [11] and viability. Due to the small size and low cost, this instrument has been attracted for field measurements of both concentration and viability. In our hand this instrument was also became useful for the quick measurement of sperm concentration an d viability in stallion (Morrell et al., 2010). Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that operates in the ultraviolet light range, are less commonly used as this is not a standard feature on the smaller analytical machines. However, one alternative is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect nonviable bull spermatozoa by fluorometry and obtained an inverse correlation between the damaged cells per cent and the field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with motility. Fluorochromes used to assess sperm viability by both approach could be utilized in combination with each other. In that case, when CFDA is used combined with PI, three populations of cells as live, which are green; dead, which are red; and a third population which is stained with both and represents dying spermatozoa can be identified. This combination was found useful by Almlid and Johnson (1988) for frozen-thawed boar spermatozoa for monitoring membrane damage at the time of evaluation of various freezing protocols. Further, Harrison and Vickers (1990) also noticed that this combination with a fluorescent microscope is effective indicator of viability of fresh, incubated or cold-shocked spermatozoa in boar and ram. Contrasting to these, Garner et al. (1986) was failed to find a relationship between bull sperm viability and fertility when using combination of CFDA/PI . Flow cytometry for evaluating sperm viability appears to be a precious tool in the AI industry. When a high number of sperm is packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect. However, sperm viability might be more important when combined with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combined assessment of sperm viability and concentration appears to be useful in the wake of improving quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively bright fluorescence was found also in the mitochondrial sheath of living sperm. But the mechanism and mode of action by which SYBR-14 binds to the DNA of sperm is not known. It is know that PI stains nucleic acids by inte rcalating between the base pairs (Krishan, 1975). Viability stains can also be used in conjugation with fluorescently labeled plant lectins for simultaneous assessment of the plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). It is conceivable that assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garner et al. (5) found that insemination of boar sperm stained with SYBR-14 did not compromise fertilization or even the development of flushed porcine embryos in vitro. Non-viable sperms can be detected using the membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with damaged membranes. Plasma membrane which is intact will not permit these stains entering into the spermatozoa and staining the nucleus. Most frequently used stains include phenanthridines, for example propidium iodide (PI; (Matyus, 1984) ethidium homodimer-1 (EthD-1; (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). After a series of comparison between fertility of cryopreserved stallion spermatozoa with a number of laboratory assessments of semen quality as assessed by flow cytometry using PI, Wilhelm et al. (1996) concluded that viability is the single laboratory assay that correlated with fertility. -Sperm plasma membrane integrity Although the sperm plasma membrane covers the entire cell, it consists of three distinct membrane compartments, one which covers the outer acrosomal membrane, one which covers the post acrosomal portion of the sperm head, and one which covers the middle and principal pieces. Sperm membrane is directly or ind