Practical step of seminal fluid analysis

EXAMINATION PARAMETER

MACROSCOPIC EXAMINATION

• Time of arrival
• Appearances of the specimen
• Viscosity/liquefaction
• Volume of the specimen

MICROSCOPIC EXAMINATION

MOTILITY

1. Quantitative motility
2. Qualitative motility

Sperm count

HOW TO COLLECT SAMPLE

• The sample should be collected after abstaining from sex for 2-5 days
• The sample should be collected in the laboratory by well trained laboratory health personnel via masturbation
• The sample must be delivered to the laboratory within 2 hours after collection when the sample is collected outside of the laboratory
• The used of condom is not allowed, but it can be use if he condom if free of spermicides or lubricant
• The sample should be discarded when leakage where found in the container during transportation of the sample to the laboratory
• The container must be free and sterile and the name of the patient must be fully written on it.
• The temperature range should be maintain, the average temperature should be less that 20^oC and it should not be more than 40^oC for exposure
• The examination of single specimen should not be encourage, at list two to three specimen should be collected a week or two week interval.

medical laboratory

NORMAL SEMEN CHARACTERISTIC

Freshly ejaculated semen should have the following characteristic;

• Highly viscid opaque and creamy
• White in colour and have a distinct musky or sickly smell
• The specimen should liquefy within 20 minute after collection
• On liquefaction the specimen may change slightly viscous
• The specimen may be turbid and alkaline with Ph 7.7
• The total volume is measure, which after 2-5 days abstinence from sex, the volume should be from 2-3ml

PARAMETER PROCEDURE

1 TIME OF ARIVAL

2 APEARANCE

3 VISCOSITY/LIQUEFACTION

PROCEDURE

A sterile pipette should be used to suck up the fluid and then release it, if the liquid drop slowly one by one, it viscosity is normal. Then report normal viscosity.

But if the fluid fallout from the pipette without dropping, then report watery semen

NB: increase viscosity of the sperm can inhibit the sperm motility.

 

4 VOLUME

Liquefy semen can be measure with 10ml or 5ml pipette. For viscosity semen, wide mouth pipette should be used.

NB: Less volume of the spermatozoa may be due to lack of abstinence from sex, of which it can also interfere with the sperm count.

MICROSCOPY EXAMINATIION

MORTILITY

 Include Quantitative motility and Qualitative motility

Normal sperm Quantitative motility is from 60% upward

LABORATORY PROCEDURE

One drop of liquefied mixed semen should be place on a clean grease free slide, cover with cover slip and examine microscopically with x10 objective lens with iris diaphragm close, then focus properly with x40 objective.

WHO (world health organization) recommend for motility analysis.

QUANTITATIVE MOTILITY

This should be determined by counting both number of motile and non motile for at list 10 randomly selected fields, and at list 100 must be counted.

The percentage of motile sperm is calculated from the means percentage motility for the entire field counted. This is then adjusted to the nearest 5% (e.g 73% = 75%).

QUALITATIVE MOTILITY

This are determine by the percentage progress movement of the sperm under examination.

The result should be reported as follow

• NON: when there are no progressive movement seen
• POOR: when the number of the progressive movement is less than non motile
• GOOD: when the number of the progressive movement is slightly higher than the number of non motile
• EXCELLENT: when almost or all the number of sperm are motile

SPERM COUNT

Sperm count is carried out with the use of haemocytometer. Also electronic coulter counter can be used if it is available.

METHOD

The specimen should be diluted in 1:20 by using WBC (white blood cell) pipette.

Draw 0.5 mark of semen dilute it with diluting fluid of 11 mark.

Alternatively; micro pipette can be used, when using micro pipette,

Measure 10µl, 50µl, or 100µ volume can be used to make 1:20 dilution in a test tube.

NB: the diluting fluid should immobilize and preserve sperms.

COMMON DILUTING FLUID COMPOSITION

1. Sodium bicarbonate (NaHCO₃) 5g
2. Formalin (neutral) 1ml
3. Distilled water      100ml

NB: this diluting is satisfactory for both bright and phase contrast microscopy.

But 0.5ml of saturated aqueous solution of gentian violet can be added for light microscopy.

 

Also tap water that is not acidic can be used for dilution.

 

                                 COUNTING PROCEDURE CHART

1									2
	5a							5b
					5e
	5c							5d
3									4

It only area label 5 should be counted, which is 5a, 5b, 5c, 5d, and 5e are the area that should counted.

CALCULATION

Sperm count = n

Multiplication factor = 50,000

Dilution factor = 20

 Total calculation will be : n x 50,000 x 20 = n x 10⁶ml