| Materials Needed The following is a list of materials needed for semen analysis. Laboratories will differ slightly in the equipment used. Use of this equipment will be described further in the later pages of this course.
Materials needed include:graduated test tube or serological pipets with safety bulb to measure volumepH paper in neutral to basic range (e.g. 7.2-8.8)counting chamber and/or automated counting machineglass slides and coverslips for wet mount if motility and sperm count are to be assessed separatelyhand counterif dilution is donediluting fluid calibrated automatic pipetspositive pressure pipets and glass boreslight microscope with phase contrast objectives for sperm count and bright field objectives for morphology assessmentglass slides and fixative for morphology slidesset-up for performing Papanicolaou or other morphology stainingEvery laboratory should also have a copy of the "WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction", published on behalf of the WHO by Cambridge University Press. The fourth edition was published in 1999. | View Page |
| In summary the procedure will involve the following: The following aspects of semen analysis will be described in further detail during this course: Check the identity of the patient Record information that has been obtained from the patient including: time of collection, collection method, problems during collection, medications the patient is taking Note time to liquefaction Measure the volume by pouring into a graduated test tube or by drawing the specimen into an appropriately sized graduated serological pipet Assess viscosity Note color Measure pH by putting a drop on a strip of pH paper Count the sperm in the specimen Assess motility Count round cells, if present Assess the proportion of round cells that are white cells Fix and prepare specimen for morphology assessment; assess morphology | View Page |
| Examination of semen is important because: | View Page |
| Importance of Semen Analysis Semen analysis is important for several reasons:To determine whether compromised semen parameters contribute to infertilityTo confirm that vasectomy has reduced sperm count to zeroTo confirm the presence of sperm following reversal of vasectomyTo confirm or refute the presence of sperm in cases where rape is suspected or paternity is in question | View Page |
| Limits of Semen Analysis Semen analysis can provide important information related to the function of the male reproductive system but, even when results are within normal limits, it does not ensure that a male is fertile.
A normal semen analysis result does not mean that all causes of male infertility have been ruled out. One reason for this is that there can be considerable differences between one semen analysis result and another in a single individual.
On the other hand, an abnormal result does not always mean that a couple cannot conceive a pregnancy. Men with suboptimal sperm counts have been known to father children. Also, infection, trauma, stress, febrile illness and medications can cause temporary subfertility.
For all of these reasons multiple specimens are recommended for a complete analysis of the semen.
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| Evaluating Male Infertility Semen analysis has become an important screening test when evaluating male infertility. Infertility has been defined as the failure of a couple to conceive after one year of unprotected intercourse. The results of the semen analysis may provide sufficient information for a diagnosis or may indicate the need for additional testing. | View Page |
| Purpose This course will give you an overview of the methods involved in performing a semen analysis.
Semen analysis may be performed for one of several reasons. One of these is evaluation to assess male fertility. Infertility is a problem for approximately 1 in 7 couples who attempt a first pregnancy. In almost half of these cases (~50%) the cause of infertility can be traced, at least in part, to an abnormality in the male. Examination of sperm is the first step in evaluating male infertility.
Semen analysis can also be used to: confirm the absence of sperm in post vasectomy patients; confirm the presence of sperm after vasectomy reversal; and to determine the presence of sperm for certain legal purposes, such as rape. | View Page |
| Prerequisites The basic laboratory skills that you will need to do a semen analysis include:
Using a microscopePerforming manual cell counts and doing calculations to determine the concentration of those cells per milliliter of fluidMeasuring volumeMeasuring pHMeasuring viabilityKnowledge of OSHA regulations for handling potentially infectious human fluids | View Page |
| Summary: Reference values The following are reference values for a normal semen analysis. It should be noted that these are recommended reference ranges only and that they may require adjustment for your particular laboratory or region of the country:Liquefaction: ≤30 minutesVolume: ≥2.0 mlColor: white, yellowish, grayViscosity: non-viscouspH: ≥7.0Sperm count: ≥20 million / mlMotility: ≥50%Leukocytes: ≤1 million / mlWHO III Morphology: ≥30%Strict Morphology: ≥14%
In addition some people find it useful to have a total motile count (TMC). This is calculated by multiplying the concentration x the percent motility x the volume. Normal TMC is 10 million or greater. | View Page |
| Sample semen analysis report The semen analysis report must show all of the information collected and must contain reference values for normal ranges. | View Page |
| Fructose Fructose makes up 99% of the reducing sugar present in semen. This sugar is produced in the seminal vesicles and its absence may indicate an obstruction proximal to these glands.
Although a fructose test is NOT part of a routine semen analysis, the clinician may want to measure this in cases of azoospermia. In azoospermia secondary to obstruction of the ejaculatory ducts or absence of the vas deferens, fructose is usually absent. When azoospermia is caused by failure of the testes to produce sperm, fructose is present. Measuring fructose levels can thus help the clinician determine the cause of azoospermia, although measurement of pH is often more useful in this regard.
The procedure for determining the amount of fructose in semen involves heating semen in a strong acid in the presence of resorcinol. Fructose gives a red color to this solution when present. | View Page |
| Liquefaction Immediately following ejaculation, semen is in a gel-like condition.Liquefaction, or resolution of the gel-like consistency, is expected within 15 minutes. If liquefaction does not occur within 60 minutes you should note this on the report sheet.Occasionally a specimen does not liquefy. If this occurs, mechanical mixing or enzyme treatment may be necessary in order for the sperm count, motility analysis and other microscopic aspects of semen analysis to be performed. | View Page |
| Collection Accurate semen analysis results require appropriate sample collection. Patients must receive detailed directions for proper specimen collection and transport. Directions should be in writing. Specific instructions should include: The period of abstinence prior to collection should be between 2 and 5 days.The entire specimen must be collected because the different portions have varying concentrations of spermatozoa.An appropriate collection container must be used.Each laboratory should designate an appropriate, wide mouth, collection container.Each lot of collection containers should be tested to ensure that it is non-toxic to sperm.Alternative collection containers should be discouraged because their level of toxicity is unknown.Use of condoms for collection should be discouraged particularly when the purpose of the semen analysis is to test for fertility. Some condoms are toxic to sperm. Collection in condoms often results in inaccurate results for semen volume and other parameters. | View Page |
| Identity of patient When doing a semen analysis it is important to be certain that the patient has been correctly identified prior to collection. At some locations the patient is asked to show a picture ID at the time of collection or when accepting a specimen produced at home.
In situations where rape is suspected or paternity is in question, law enforcement officials should be consulted. It may be necessary for law enforcement officials to be present to identify the patient and to ensure that chain of custody has been correctly established. | View Page |