Semen Analysis. laboratory body fluid .ppt

lab. test for body fluid in seminal analysis

1. 1
Semen Semen
An
alysis
An
alysis
Dr. Amal BaalashDr. Amal Baalash

2. IndicationsIndications

Assessment of fertilityAssessment of fertility

Forensic purposesForensic purposes

Effectiveness of vasectomy - 2 samples 1 Effectiveness of vasectomy - 2 samples 1
month apart negativemonth apart negative

Suitability for artificial inseminationSuitability for artificial insemination

3. Semen Analysis IncludeSemen Analysis Include

MacroscopicMacroscopic

viscosityviscosity

coagulation + coagulation +
liquifactionliquifaction

volumevolume

pHpH

MicroscopicMicroscopic

concentration/concentration/
countcount

motilitymotility

morphologymorphology

viabilityviability
Motility &Viability must be performed within 1½ - 2 Motility &Viability must be performed within 1½ - 2
hrs of collectionhrs of collection

4. REMEMBERREMEMBER
SEMEN IS A BODY FLUIDSEMEN IS A BODY FLUID
BIOHAZARDOUSBIOHAZARDOUS

5. Semen CollectionSemen Collection

Name Name

Period of abstinence - 2-7daysPeriod of abstinence - 2-7days

Time of collection Time of collection

Entire ejaculate and not coitus interruptus in Entire ejaculate and not coitus interruptus in
a wide mouth containera wide mouth container

Delivered within 1 hour of collectionDelivered within 1 hour of collection

Avoid temperature extremesAvoid temperature extremes

6. Reference RangesReference Ranges

Volume 2.0-6.0 mlVolume 2.0-6.0 ml

pH 7.2-8.0pH 7.2-8.0

Count >20 million/mlCount >20 million/ml

Total count > 40 millionTotal count > 40 million

Morphology > 30% normal Morphology > 30% normal
formform

Viability > 75%(50% in other)Viability > 75%(50% in other)

WBC< 1million/mlWBC< 1million/ml

RBC noneRBC none

7. Macroscopic ExaminationMacroscopic Examination

Semen is viscous, yellow grayish.Semen is viscous, yellow grayish.

Forms gel-like clot immediately.Forms gel-like clot immediately.

Liquefies completely in 5-60 minutes; this Liquefies completely in 5-60 minutes; this
must be complete before further testing must be complete before further testing
(mix before further testing).(mix before further testing).

Appearance: homogenous white-gray Appearance: homogenous white-gray
opalescence.opalescence.

Brown/red in hematospermiaBrown/red in hematospermia

Dense white turbid if inflammation and high Dense white turbid if inflammation and high
WBCWBC

8. 
Volume: in graduated cylinder to the Volume: in graduated cylinder to the
nearestnearest 0.1 ml or centrifuge tube free of 0.1 ml or centrifuge tube free of
contamination.contamination.

Viscosity: 5ml pipette or plastic pipetteViscosity: 5ml pipette or plastic pipette

normal, more viscous, very viscousnormal, more viscous, very viscous

pH: important parameter of motility and pH: important parameter of motility and
viabilityviability7.2-8.0; measured by pH paper. 7.2-8.0; measured by pH paper.
Macroscopic ExaminationMacroscopic Examination

9. MotilityMotility

While estimating countWhile estimating count

No stainNo stain

Count 200 total sperm and then the motileCount 200 total sperm and then the motile

Calculate the percentage ofCalculate the percentage of

Progressive motile Progressive motile

Sluggishly motile (<5 um/s )Sluggishly motile (<5 um/s )

nonmotilenonmotile

>50% motile>50% motile

11. 11

13. AgglutinationAgglutination

Reported when Reported when motilemotile sperm stick to sperm stick to
each other in a definite pattern.each other in a definite pattern.

Head-headHead-head

Tail-tailTail-tail

Head-tailHead-tail

Immunological cause of infertilityImmunological cause of infertility

Done on several HPFDone on several HPF

14. ViabilityViability

Supravital stain: Supravital stain:

Eosin +/- NigrosinEosin +/- Nigrosin

Viable do not take up the stainViable do not take up the stain

This distinguish live nonmotile from This distinguish live nonmotile from
dead; it is important to compare dead; it is important to compare
viability and motility.viability and motility.

15. MorphologyMorphology

Smear: Smear:

H&E, Papanicolaou, Wright stainsH&E, Papanicolaou, Wright stains

Feathering like blood smear or 2 slidesFeathering like blood smear or 2 slides

Count and classify 100-200 spermatozoaCount and classify 100-200 spermatozoa

Examine the head, midpiece, tailExamine the head, midpiece, tail

Normal >30%Normal >30%

ImmatureImmature

AbnormalAbnormal

16. Mira1000 Semen Analyzer Mira1000 Semen Analyzer
(CASA)(CASA)
16

17. 
Aspermia:Aspermia: No semen ejaculated No semen ejaculated

Hematospermia:Hematospermia: Blood present in semen Blood present in semen

LeucocytospermiaLeucocytospermia:: White blood cells present in White blood cells present in
semen semen

Azospermia:Azospermia: No spermatozoa found in semen No spermatozoa found in semen

Normospermia:Normospermia: Normal semen parameters Normal semen parameters

Oligospermia:Oligospermia: Low sperm concentration Low sperm concentration

Asthenospermia:Asthenospermia: Poor motility and/or forward Poor motility and/or forward
progression progression

Teratospermia:Teratospermia: Reduced percentage of Reduced percentage of
morphologicall normal sperm morphologicall normal sperm

Necrospermia:Necrospermia: No live sperm in semen No live sperm in semen

18. 
Head abnormalities:Head abnormalities:

absence absence

double headdouble head

micro/megalomicro/megalo
Other Sperm AbnormalitiesOther Sperm Abnormalities

Tail abnormalities:Tail abnormalities:

coiledcoiled

kinkedkinked

lengthenedlengthened

20. 20

21. Sperm CountSperm Count

Manual methodsManual methods

Hemocytometer or counting chamberHemocytometer or counting chamber

Computer assisted Computer assisted

Oligospermia<20 millionOligospermia<20 million

If azospermia: fructose level must be If azospermia: fructose level must be
ordered to verify the integrity of the vas ordered to verify the integrity of the vas
and seminal vesiclesand seminal vesicles

22. PreparationPreparation

Manual methodsManual methods

Hemocytometer or counting chamberHemocytometer or counting chamber

Computer assistedComputer assisted
1. Thoroughly mix specimen and dilute 1:10 with 1. Thoroughly mix specimen and dilute 1:10 with
diluent. (To obtain this dilution, dilute 100 uL of diluent. (To obtain this dilution, dilute 100 uL of
liquefied semen with 900 uL of diluent)liquefied semen with 900 uL of diluent)
2. Thoroughly mix diluted specimen and allow a 2. Thoroughly mix diluted specimen and allow a
drop (10 - 20 uL) to into each side of the drop (10 - 20 uL) to into each side of the
hemocytometer covered with a coverglass.hemocytometer covered with a coverglass.
3. Allow chamber to stand for about 5 minutes in a 3. Allow chamber to stand for about 5 minutes in a
humid container to prevent drying. During this humid container to prevent drying. During this
period, the cells settle and can be more easily period, the cells settle and can be more easily
counted.counted.

23. 4. After cells have settled, place chamber under a 4. After cells have settled, place chamber under a
microscope microscope
5.5.Count spermatozoa present in 5 1/25mm squares Count spermatozoa present in 5 1/25mm squares
in center square millimeter X5 or sperms in one of in center square millimeter X5 or sperms in one of
the 9 large squares. the 9 large squares.
Only morphologically mature germinal cells with tails Only morphologically mature germinal cells with tails
are counted. are counted.
No of sperms per large square X dilution factor (10) X No of sperms per large square X dilution factor (10) X
Depth of chamber (10) X 1000 = count in million/ mlDepth of chamber (10) X 1000 = count in million/ ml
PreparationPreparation

24. 24

25. 
Decreased:Decreased:

vasectomy (should be 0 after 3-6 months)vasectomy (should be 0 after 3-6 months)

varicocelevaricocele

primary testicular failure (Klinefelters)primary testicular failure (Klinefelters)

secondary testicular failuresecondary testicular failure

congenital vas obstructioncongenital vas obstruction

retrograde ejaculationretrograde ejaculation

endocrine causes (prolactinemia, low testosterone)endocrine causes (prolactinemia, low testosterone)
Sperm CountSperm Count