SlideShare uma empresa Scribd logo
1 de 30
Baixar para ler offline
SEMEN ANALYSIS
DR.RENUKADEVI
SENIOR EMBRYOLOGIST
ARC International Fertility & Research
Centre
Introduction
 A semen analysis measures the amount of semen a man
produces and determines the number and quality of sperm
in the semen sample.
 A semen analysis is usually on e of the first test done to
help determine whether a man has problem fathering a child
(infertility)
 A problem with the semen or sperm affects more than one-
third of the couples who are unable to have children
(infertile)
Semen Analysis
 SA are mainly carried out to determine whether:-
 A man has a reproductive problem that is causing infertility.
 A vasectomy or vasectomy reversal has been successful
 SA is the singular test for fertility in male that can provide
information on
◦ sperm production.
◦ patency of the male ducts.
◦ the function of the accessory glands.
◦ and ejaculative function.
Since semen samples may vary from day to day, 2 or 3 samples may
be evaluated within a 3-6 month period for accurate testing.
A semen analysis to test the effectiveness of a vasectomy is usually
done 6 weeks after the vasectomy
Semen collection
 Masturbation, directing the semen into a clean sample cup. Do
not use a lubricant which can kill sperms
 Coitus interruptus - withdrawing the penis from the partner just
before ejaculating follow by ejaculating into a clean sample cup.
 Coitus - by using a condom. A special (silicon) condom that
does not contain any substance that kills sperm (spermicide).
After ejaculation, carefully remove the condom from the penis.
Tie a knot in the open end of the condom and place it in a
container that can be sealed in case the condom leaks or breaks.
Ordinary condoms should not be used since they usually contain
spermicides
 Assisted ejaculation – electro-ejaculation used in paralegics
 Once after collecting the semen sample from the patient,
receiving time is noted down.
 The sample has to be liquefied for 20-30mins at 37°c by
placing the sample in heating block. (Avoid crystallization
by leaving the sample stationary for a too long time).
 After 20-30mins of liquefaction, mix the sample
homogeneously by using transfer pipette.
 Sample volume, viscosity and pH should be noted.
SEMEN ANALYSIS
Semen Collection
• Abstinence days 2-5
• Pass urine
• Wash hands with soap, dry
• Collect the entire sample into
the wide mouth sterile container,
70% of sperms is in the first
part of the ejaculate
• Keep the sample at body
temperature, no sunlight
• Deliver the sample within one
hour of ejaculation
Good and reliable SA results starts from semen collection, preferably by
masturbation
Parameters (WHO criteria 2010)
Parameters agreed on consensus
Parameter Lower Reference Limit
Semen volume (ml) 1.5
Sperm concentration (106/ml) 15
Total sperm number (106/ejaculate) 39
Progressive motility (PR, %) 32
Total motility (PR +NP, %) 40
Vitality (live sperms, %) 58
Sperm morphology (NF, %) 4
pH* >/=7.2
Leucocyte* (106/ml) <1
MAR/Immunobead test* (%) <50
Terminologies of Semen Analysis
 Oligospermia – sperm concentration <15 million/ml
 Asthenozoospermia – <40% grade (PR+NP) or < 32 PR%
 Teratozoospermia – <4% spermatozoa
 OAT =Oligo-astheno-teratozoospermia
 Azoospermia – no spermatozoa in semen
 Polyzoospermia – ++ high sperm concentration, >200M/ml
 Hypospermia – semen volume < 1.5 ml
 Hyperspermia – semen volume > 4.5 ml
 Aspermia – no semen volume
 Pyospermia – leukocytes present in semen, >1M/ml
 Hematospermia – red blood cell present in semen
 Necrozoospermia – “dead” sperm
Macroscopic Examination
WHO criteria 2010 Description
Appearance Normal: Whitish to grey opalescent
Yellow (urine, jaundice); Pink/Reddish/Brown (RBCs)
Liquefaction Normal: 15–30 minutes after collection
Lumpy >60 min – sperms may be trapped in unliquefied
jelly; maybe sign of prostatic infection, lack of prostatic
protease
Viscosity Normal Smooth and watery
Abnormal –, thick with long threads (21G needle). High
viscosity impede sperm movements
MIX SEMEN THOROUGHLY BEFORE ANALYSIS –USE A
PIPETTE
Macroscopic Examination
Volume Normal: 1.5 ml per ejaculation
Low volume (<1ml) reflect a problem with the seminal
vesicles and prostate – a block, retrograde ejaculation,
infection or lack of androgen.
Low semen volume cannot neutralize vaginal acidity
High semen volume dilute sperms/ active infection
pH Normal: =/>7.2 (alkaline)
Acidic pH (<7.0) in a low volume & density sample
indicates –congenital bilateral absence of vas deferens (in
which seminal vesicles are also poorly developed) and
ejaculatory duct obstruction. pH increases with time as
natural buffering capacity of semen decreases – therefore
high ph is not clinically useful
Semen analysis by Dr.Renukadevi
Semen analysis by Dr.Renukadevi
Semen analysis by Dr.Renukadevi
 A microscope objective of 20x should be used for counting.
 Place a drop of the sample into Makler chamber and count the
sperm heads as follows: 10 squares of the field. This number
represents the concentration of spermatozoa in millions per
milliliter.
 When counting a sample of oligozoospermia (less than
15million/ml) all 100 squares have to be counted. Adding "00000"
to that number, you will get the concentration of the spermatozoa
per milliliter.
SPERM COUNT
Microscopic Examination
 Grade A (fast progressive) sperms are those which swim forward fast
in a straight line - like guided missiles.
 Grade B (slow progressive) sperms swim forward, but either in
a curved or crooked line, or slowly (slow linear or non linear motility).
 Grade C (non-progressive) sperms move their tails, but do not move
forward (local motility only).
 Grade D (immotile ) sperms do not move at all.
 Immotile spermatozoa are to be counted in a defined number of
squares. Afterwards the motile spermatozoa are counted and
classified into four categories (from A "fast progressive" to D "no
motility"). This procedure is to be repeated in different areas of the
grid to have the quality and motility to be calculated on base of this
data in percent.
Motility is graded from A to D,
Semen analysis by Dr.Renukadevi
Semen analysis by Dr.Renukadevi
Semen analysis by Dr.Renukadevi
Semen analysis by Dr.Renukadevi
SPERM MORPHOLOGY
A drop of sample is placed in the sterile glass slide and covered using a
cover slip, name and SID number should be mentioned. The sample is
viewed under light microscope with a magnification of 40x.
A Normal Sperm has:
A smooth, oval-shaped head that is 5-6 micrometers long and 2.5-3.5
micrometers wide (less than the size of a needle point)
A well-defined cap (acrosome) that covers 40% to 60% of the sperm
head
No visible abnormality of neck, mid-piece, or tail
No fluid droplets in the sperm head that are bigger than one half of
the sperm head size.
Semen analysis by Dr.Renukadevi
The following categories of defects should be noted.
Head defects, namely large, small, tapered, pyriform,
globe, amorphous heads, vacuolated heads, double heads and
small acrosome.
Neck and mid-piece defects namely bent neck, abnormal
mid-piece (thin or thick).
Tail defects namely short, multiple, hairpin, broken, bent
tails, coiled tail.
Multiple defects
Agglutination
Pus cells/HPF
Debris
After completion of analysis the results should be noted in
the record and typed in the patient report entry column.
Morphology assessment
 Sperms must be stained for an accurate assessment
 Strict criteria /Kruger /Tygerberg
 The sperm head is oval, smooth-symmetrical outline, a length of 3-5 um and a
width of 2-3um.
 The head should have a well defined acrosome area of 40-70% and vacuoles
(=/<2) that occupies ~ 20% head area.
 The mid-piece must be straight and slender, 0.5 um in width and 7-8um long,
straightly aligned to the head.
 The tail must be straight and 45-50 um long.
 To be classified as normal, the sperm must be normal in all portions (head,
mid-piece, tail).
 At least 400 sperms must be scored on randomly chosen fields.
Normal Forms (%) = normal sperms / the total number of sperms
evaluated x 100.
Diff-Quick Staining
 Simple, quick staining method.
 Moderate differentiation of structures --the acrosome is stained red and the
post acrosomal area dark red. Very thin smears are necessary as the
background is influenced by the presence of protein in the seminal fluid
and tends to be dark.
 The slides are fixed with fixative (Cytospray, Kinetik, Australia).
 Diff Quik 1 for 5-6 seconds.
 Clean water x 2. Removed excess moisture.
 Diff Quik 2 for 10 seconds
 2-3 dips in clean water.
 It is left to dry by standing the slide on one end on absorbent paper towel.
 Sperms stained by Diff-Quik are larger by comparison to H&E staining as
the cells did not undergo dehydration by alcohol.
Abnormal Sperms
Abnormal Sperms
1. Triple head
sperm
2. Acrosome
reacted sperm
3. Sperm with no
acrosome
4. Sperm with a
tapering head
and swollen
mid-piece
Morphology Assessment
 Assessment is subjective due to the
wide variation in sperm sizes and
shapes.
 % NF correlated well with the
fertilization rate in-vitro and
pregnancy rate [Kruger et al, 1996;
Morgenthaler et al, 1995]
 Sperms with defective heads are
more likely to be immotile than
sperms without defects, and
defective motile sperms tend to
show sluggish motility as
compared to normal sperms
[Aitken et al, 1995].
Vitality Assessment
1. Eosin-nigrosin (dead sperm stain pink/red)
2. Eosin (1%) (dead sperm stain pink/red)
3. Trypan (0.4%) blue (dead sperm stain blue)
4. Hypo-osmotic swelling test (HOS) (live sperm shows tail
curling
Test 1, 2 and 3 for diagnostic uses.
 Usually 1:1 ratio of semen to dye mixture, mix well and smear
onto a slide. Read immediately at x40 objective, count 200
sperms
Test 4 is use to choose live (immotile) sperm for ICSI
 Dead sperms will not react in HOS while live sperm will take
up fluids causing their tails to curl within 5 min and stabilize at
30 min. Therefore viable sperms may be selected for ICSI [Lin
et al, 1998; Cayan et al, 2001].
Vitality Assessment
Dead sperms are stained
pink/red
Live sperms shows curling
tails
Semen analysis by Dr.Renukadevi

Mais conteúdo relacionado

Mais procurados (20)

introduction of cytopathology
introduction of cytopathologyintroduction of cytopathology
introduction of cytopathology
 
Semen analysis dr kamlesh
Semen analysis   dr kamleshSemen analysis   dr kamlesh
Semen analysis dr kamlesh
 
Semen analysis
Semen analysisSemen analysis
Semen analysis
 
Semen examination
Semen examinationSemen examination
Semen examination
 
Hormonal cytology
Hormonal cytologyHormonal cytology
Hormonal cytology
 
Preparation and staining of peripheral blood smear
Preparation and staining of peripheral blood smearPreparation and staining of peripheral blood smear
Preparation and staining of peripheral blood smear
 
Coomb's test
Coomb's testCoomb's test
Coomb's test
 
Semen analysis WHO 2010 BY DR ANAMIKA DEV
Semen analysis WHO 2010 BY DR ANAMIKA DEVSemen analysis WHO 2010 BY DR ANAMIKA DEV
Semen analysis WHO 2010 BY DR ANAMIKA DEV
 
Blood grouping
Blood groupingBlood grouping
Blood grouping
 
semen analysis
semen analysissemen analysis
semen analysis
 
Hemoglobin estimation
Hemoglobin estimationHemoglobin estimation
Hemoglobin estimation
 
STOOL EXAMINATION
STOOL EXAMINATIONSTOOL EXAMINATION
STOOL EXAMINATION
 
Liquid based cytology
Liquid based cytologyLiquid based cytology
Liquid based cytology
 
Body fluid analysis
Body fluid analysisBody fluid analysis
Body fluid analysis
 
coombs test
coombs testcoombs test
coombs test
 
Semen analysis Latest WHO (2010)
Semen analysis Latest WHO (2010)Semen analysis Latest WHO (2010)
Semen analysis Latest WHO (2010)
 
Semen Analysis
Semen AnalysisSemen Analysis
Semen Analysis
 
Bleeding time and clotting time
Bleeding time and clotting timeBleeding time and clotting time
Bleeding time and clotting time
 
SEMEN ANALYSIS
SEMEN ANALYSISSEMEN ANALYSIS
SEMEN ANALYSIS
 
CSF Examination
CSF ExaminationCSF Examination
CSF Examination
 

Semelhante a Semen analysis by Dr.Renukadevi

Semelhante a Semen analysis by Dr.Renukadevi (20)

Iui workshop femelife
Iui workshop femelifeIui workshop femelife
Iui workshop femelife
 
chapter 4.pptx
chapter 4.pptxchapter 4.pptx
chapter 4.pptx
 
Recent advances in male infertility
Recent advances in male infertilityRecent advances in male infertility
Recent advances in male infertility
 
Sperm preparation by Dr.Renukadevi
Sperm preparation by Dr.RenukadeviSperm preparation by Dr.Renukadevi
Sperm preparation by Dr.Renukadevi
 
SEMEN ANALYSIS.pptx
SEMEN ANALYSIS.pptxSEMEN ANALYSIS.pptx
SEMEN ANALYSIS.pptx
 
Semen analysis or seminal fluid analysis
Semen analysis or seminal fluid analysisSemen analysis or seminal fluid analysis
Semen analysis or seminal fluid analysis
 
Semen analysis
Semen analysisSemen analysis
Semen analysis
 
SEMEN ANALYSIS PPT.pptx
SEMEN ANALYSIS PPT.pptxSEMEN ANALYSIS PPT.pptx
SEMEN ANALYSIS PPT.pptx
 
SEMEN ANALYSIS POWERPOINT
SEMEN ANALYSIS POWERPOINTSEMEN ANALYSIS POWERPOINT
SEMEN ANALYSIS POWERPOINT
 
Infertility and Sperm analysis
Infertility and Sperm analysisInfertility and Sperm analysis
Infertility and Sperm analysis
 
SEMEN EVALUATION
SEMEN EVALUATIONSEMEN EVALUATION
SEMEN EVALUATION
 
sperm assessment- traditional and novel approaches.pptx
sperm assessment- traditional and novel approaches.pptxsperm assessment- traditional and novel approaches.pptx
sperm assessment- traditional and novel approaches.pptx
 
Male infertility investigations-Dr.Vishnu Bawane
Male infertility investigations-Dr.Vishnu BawaneMale infertility investigations-Dr.Vishnu Bawane
Male infertility investigations-Dr.Vishnu Bawane
 
semen analysis.ppt
semen analysis.pptsemen analysis.ppt
semen analysis.ppt
 
histo semen.pptx
histo semen.pptxhisto semen.pptx
histo semen.pptx
 
Semen Analysis
Semen AnalysisSemen Analysis
Semen Analysis
 
Andrology lab
Andrology labAndrology lab
Andrology lab
 
Dog vaginal cytology by Dr.mehdi moradi
Dog vaginal cytology by Dr.mehdi moradiDog vaginal cytology by Dr.mehdi moradi
Dog vaginal cytology by Dr.mehdi moradi
 
Fertility tests
Fertility testsFertility tests
Fertility tests
 
Histological and cytological specimens
Histological and cytological specimensHistological and cytological specimens
Histological and cytological specimens
 

Mais de Morris Jawahar

An overview of IVF/ICSI by Dr.Renukadevi
An overview of IVF/ICSI by Dr.RenukadeviAn overview of IVF/ICSI by Dr.Renukadevi
An overview of IVF/ICSI by Dr.RenukadeviMorris Jawahar
 
Sperm DNA fragmentation by Dr.Renukadevi
Sperm DNA fragmentation by Dr.RenukadeviSperm DNA fragmentation by Dr.Renukadevi
Sperm DNA fragmentation by Dr.RenukadeviMorris Jawahar
 
An overview of IVF/ICSI
An overview of IVF/ICSIAn overview of IVF/ICSI
An overview of IVF/ICSIMorris Jawahar
 
Thyroid and fertility by Dr. Gayathiri
Thyroid and fertility by Dr. GayathiriThyroid and fertility by Dr. Gayathiri
Thyroid and fertility by Dr. GayathiriMorris Jawahar
 
Overview of infertility by Dr.Gayathiri
Overview of infertility by Dr.GayathiriOverview of infertility by Dr.Gayathiri
Overview of infertility by Dr.GayathiriMorris Jawahar
 
History taking in infertility by Dr. Gayathiri
History taking in infertility by Dr. GayathiriHistory taking in infertility by Dr. Gayathiri
History taking in infertility by Dr. GayathiriMorris Jawahar
 
Spermcryoperservation by Dr.Chandan
Spermcryoperservation by Dr.Chandan Spermcryoperservation by Dr.Chandan
Spermcryoperservation by Dr.Chandan Morris Jawahar
 
Tubal factor and fertility by Dr.Gayathiri
Tubal factor and fertility by Dr.GayathiriTubal factor and fertility by Dr.Gayathiri
Tubal factor and fertility by Dr.GayathiriMorris Jawahar
 
Advanced reproductive age and fertility by Dr. Gayathiri
Advanced reproductive age and fertility by Dr. GayathiriAdvanced reproductive age and fertility by Dr. Gayathiri
Advanced reproductive age and fertility by Dr. GayathiriMorris Jawahar
 
Setting up of art lab by Dr. Renukadevi
Setting up of art lab by Dr. RenukadeviSetting up of art lab by Dr. Renukadevi
Setting up of art lab by Dr. RenukadeviMorris Jawahar
 
Scoring of embryos by Dr.Renukadevi
Scoring of embryos by Dr.RenukadeviScoring of embryos by Dr.Renukadevi
Scoring of embryos by Dr.RenukadeviMorris Jawahar
 
Third party reproduction ppt by Dr.Gayathiri
Third party reproduction ppt by Dr.GayathiriThird party reproduction ppt by Dr.Gayathiri
Third party reproduction ppt by Dr.GayathiriMorris Jawahar
 
Surrogacy by Dr. Gayathiri
Surrogacy by Dr. GayathiriSurrogacy by Dr. Gayathiri
Surrogacy by Dr. GayathiriMorris Jawahar
 
Azoospermia by Dr.Saravanan
Azoospermia by Dr.SaravananAzoospermia by Dr.Saravanan
Azoospermia by Dr.SaravananMorris Jawahar
 
Ivf stimulation protocols by Dr. Mahalakshmi Saravanan
Ivf stimulation protocols by Dr. Mahalakshmi SaravananIvf stimulation protocols by Dr. Mahalakshmi Saravanan
Ivf stimulation protocols by Dr. Mahalakshmi SaravananMorris Jawahar
 
Stem cells and infertility by Dr. Gayathiri
Stem cells and infertility by Dr. GayathiriStem cells and infertility by Dr. Gayathiri
Stem cells and infertility by Dr. GayathiriMorris Jawahar
 
Endometrial receptivity assay, by Dr.Gayathiri
Endometrial receptivity assay, by Dr.Gayathiri Endometrial receptivity assay, by Dr.Gayathiri
Endometrial receptivity assay, by Dr.Gayathiri Morris Jawahar
 

Mais de Morris Jawahar (17)

An overview of IVF/ICSI by Dr.Renukadevi
An overview of IVF/ICSI by Dr.RenukadeviAn overview of IVF/ICSI by Dr.Renukadevi
An overview of IVF/ICSI by Dr.Renukadevi
 
Sperm DNA fragmentation by Dr.Renukadevi
Sperm DNA fragmentation by Dr.RenukadeviSperm DNA fragmentation by Dr.Renukadevi
Sperm DNA fragmentation by Dr.Renukadevi
 
An overview of IVF/ICSI
An overview of IVF/ICSIAn overview of IVF/ICSI
An overview of IVF/ICSI
 
Thyroid and fertility by Dr. Gayathiri
Thyroid and fertility by Dr. GayathiriThyroid and fertility by Dr. Gayathiri
Thyroid and fertility by Dr. Gayathiri
 
Overview of infertility by Dr.Gayathiri
Overview of infertility by Dr.GayathiriOverview of infertility by Dr.Gayathiri
Overview of infertility by Dr.Gayathiri
 
History taking in infertility by Dr. Gayathiri
History taking in infertility by Dr. GayathiriHistory taking in infertility by Dr. Gayathiri
History taking in infertility by Dr. Gayathiri
 
Spermcryoperservation by Dr.Chandan
Spermcryoperservation by Dr.Chandan Spermcryoperservation by Dr.Chandan
Spermcryoperservation by Dr.Chandan
 
Tubal factor and fertility by Dr.Gayathiri
Tubal factor and fertility by Dr.GayathiriTubal factor and fertility by Dr.Gayathiri
Tubal factor and fertility by Dr.Gayathiri
 
Advanced reproductive age and fertility by Dr. Gayathiri
Advanced reproductive age and fertility by Dr. GayathiriAdvanced reproductive age and fertility by Dr. Gayathiri
Advanced reproductive age and fertility by Dr. Gayathiri
 
Setting up of art lab by Dr. Renukadevi
Setting up of art lab by Dr. RenukadeviSetting up of art lab by Dr. Renukadevi
Setting up of art lab by Dr. Renukadevi
 
Scoring of embryos by Dr.Renukadevi
Scoring of embryos by Dr.RenukadeviScoring of embryos by Dr.Renukadevi
Scoring of embryos by Dr.Renukadevi
 
Third party reproduction ppt by Dr.Gayathiri
Third party reproduction ppt by Dr.GayathiriThird party reproduction ppt by Dr.Gayathiri
Third party reproduction ppt by Dr.Gayathiri
 
Surrogacy by Dr. Gayathiri
Surrogacy by Dr. GayathiriSurrogacy by Dr. Gayathiri
Surrogacy by Dr. Gayathiri
 
Azoospermia by Dr.Saravanan
Azoospermia by Dr.SaravananAzoospermia by Dr.Saravanan
Azoospermia by Dr.Saravanan
 
Ivf stimulation protocols by Dr. Mahalakshmi Saravanan
Ivf stimulation protocols by Dr. Mahalakshmi SaravananIvf stimulation protocols by Dr. Mahalakshmi Saravanan
Ivf stimulation protocols by Dr. Mahalakshmi Saravanan
 
Stem cells and infertility by Dr. Gayathiri
Stem cells and infertility by Dr. GayathiriStem cells and infertility by Dr. Gayathiri
Stem cells and infertility by Dr. Gayathiri
 
Endometrial receptivity assay, by Dr.Gayathiri
Endometrial receptivity assay, by Dr.Gayathiri Endometrial receptivity assay, by Dr.Gayathiri
Endometrial receptivity assay, by Dr.Gayathiri
 

Último

Health literacies in marginalised communities LILAC 24.pptx
Health literacies in marginalised communities LILAC 24.pptxHealth literacies in marginalised communities LILAC 24.pptx
Health literacies in marginalised communities LILAC 24.pptxPamela McKinney
 
FINAL PROJECT IN EMPOWERMENT TECHNOLOGIES 11
FINAL PROJECT IN EMPOWERMENT TECHNOLOGIES  11FINAL PROJECT IN EMPOWERMENT TECHNOLOGIES  11
FINAL PROJECT IN EMPOWERMENT TECHNOLOGIES 11crzljavier
 
Assisted Living Care Residency - PapayaCare
Assisted Living Care Residency - PapayaCareAssisted Living Care Residency - PapayaCare
Assisted Living Care Residency - PapayaCareratilalthakkar704
 
2024 Compliatric Webianr Series - Contracts and MOUs from a HRSA Perspective.pdf
2024 Compliatric Webianr Series - Contracts and MOUs from a HRSA Perspective.pdf2024 Compliatric Webianr Series - Contracts and MOUs from a HRSA Perspective.pdf
2024 Compliatric Webianr Series - Contracts and MOUs from a HRSA Perspective.pdfCompliatric Where Compliance Happens
 
person with disability and pwd act ppt.pptx
person with disability and pwd act ppt.pptxperson with disability and pwd act ppt.pptx
person with disability and pwd act ppt.pptxMUKESH PADMANABHAN
 
Identifying Signs of Mental Health Presentation (1).pptx
Identifying Signs of Mental Health Presentation (1).pptxIdentifying Signs of Mental Health Presentation (1).pptx
Identifying Signs of Mental Health Presentation (1).pptxsandhulove46637
 
21 NEMT Trends & Statistics to Know in 2024
21 NEMT Trends & Statistics to Know in 202421 NEMT Trends & Statistics to Know in 2024
21 NEMT Trends & Statistics to Know in 2024Traumasoft LLC
 
EYE CANCER.pptx prepared by Neha kewat digital learning
EYE CANCER.pptx prepared by  Neha kewat digital learningEYE CANCER.pptx prepared by  Neha kewat digital learning
EYE CANCER.pptx prepared by Neha kewat digital learningNehaKewat
 
NEONATAL RESPIRATORY CARE FROM A PHYSIO POV.pptx
NEONATAL RESPIRATORY CARE FROM A PHYSIO POV.pptxNEONATAL RESPIRATORY CARE FROM A PHYSIO POV.pptx
NEONATAL RESPIRATORY CARE FROM A PHYSIO POV.pptxHanineHassan2
 
Understanding Warts and Moles: Differences, Types, and Common Locations
Understanding Warts and Moles: Differences, Types, and Common LocationsUnderstanding Warts and Moles: Differences, Types, and Common Locations
Understanding Warts and Moles: Differences, Types, and Common LocationsNeha Sharma
 
ARTHRITIS.pptx Prepared by monika gopal Tutor
ARTHRITIS.pptx Prepared  by monika gopal TutorARTHRITIS.pptx Prepared  by monika gopal Tutor
ARTHRITIS.pptx Prepared by monika gopal TutorNehaKewat
 
Three Keys to a Successful Margin: Charges, Costs, and Labor
Three Keys to a Successful Margin: Charges, Costs, and LaborThree Keys to a Successful Margin: Charges, Costs, and Labor
Three Keys to a Successful Margin: Charges, Costs, and LaborHealth Catalyst
 
Artificial Intelligence: Diabetes Management
Artificial Intelligence: Diabetes ManagementArtificial Intelligence: Diabetes Management
Artificial Intelligence: Diabetes ManagementIris Thiele Isip-Tan
 
Basics of Giant Cell Tumor of bone (GCTB)
Basics of Giant Cell Tumor of bone (GCTB)Basics of Giant Cell Tumor of bone (GCTB)
Basics of Giant Cell Tumor of bone (GCTB)bishwabandhuniraula
 
LARYNGEAL CANCER.pptx Prepared by Neha Kewat
LARYNGEAL CANCER.pptx  Prepared by Neha KewatLARYNGEAL CANCER.pptx  Prepared by Neha Kewat
LARYNGEAL CANCER.pptx Prepared by Neha KewatNehaKewat
 
Living Well Every Day: Lyons Wellness Practice | Nurtures Your Complete Health
Living Well Every Day: Lyons Wellness Practice | Nurtures Your Complete HealthLiving Well Every Day: Lyons Wellness Practice | Nurtures Your Complete Health
Living Well Every Day: Lyons Wellness Practice | Nurtures Your Complete HealthLyons Health
 
Diseases of the Respiratory System (J00-J99),.pptx
Diseases of the Respiratory System (J00-J99),.pptxDiseases of the Respiratory System (J00-J99),.pptx
Diseases of the Respiratory System (J00-J99),.pptxEMADABATHINI PRABHU TEJA
 

Último (20)

Health literacies in marginalised communities LILAC 24.pptx
Health literacies in marginalised communities LILAC 24.pptxHealth literacies in marginalised communities LILAC 24.pptx
Health literacies in marginalised communities LILAC 24.pptx
 
FINAL PROJECT IN EMPOWERMENT TECHNOLOGIES 11
FINAL PROJECT IN EMPOWERMENT TECHNOLOGIES  11FINAL PROJECT IN EMPOWERMENT TECHNOLOGIES  11
FINAL PROJECT IN EMPOWERMENT TECHNOLOGIES 11
 
Assisted Living Care Residency - PapayaCare
Assisted Living Care Residency - PapayaCareAssisted Living Care Residency - PapayaCare
Assisted Living Care Residency - PapayaCare
 
SCOPE OF CRITICAL CARE ORGANIZATION
SCOPE OF CRITICAL CARE ORGANIZATIONSCOPE OF CRITICAL CARE ORGANIZATION
SCOPE OF CRITICAL CARE ORGANIZATION
 
2024 Compliatric Webianr Series - Contracts and MOUs from a HRSA Perspective.pdf
2024 Compliatric Webianr Series - Contracts and MOUs from a HRSA Perspective.pdf2024 Compliatric Webianr Series - Contracts and MOUs from a HRSA Perspective.pdf
2024 Compliatric Webianr Series - Contracts and MOUs from a HRSA Perspective.pdf
 
person with disability and pwd act ppt.pptx
person with disability and pwd act ppt.pptxperson with disability and pwd act ppt.pptx
person with disability and pwd act ppt.pptx
 
Identifying Signs of Mental Health Presentation (1).pptx
Identifying Signs of Mental Health Presentation (1).pptxIdentifying Signs of Mental Health Presentation (1).pptx
Identifying Signs of Mental Health Presentation (1).pptx
 
21 NEMT Trends & Statistics to Know in 2024
21 NEMT Trends & Statistics to Know in 202421 NEMT Trends & Statistics to Know in 2024
21 NEMT Trends & Statistics to Know in 2024
 
EYE CANCER.pptx prepared by Neha kewat digital learning
EYE CANCER.pptx prepared by  Neha kewat digital learningEYE CANCER.pptx prepared by  Neha kewat digital learning
EYE CANCER.pptx prepared by Neha kewat digital learning
 
NEONATAL RESPIRATORY CARE FROM A PHYSIO POV.pptx
NEONATAL RESPIRATORY CARE FROM A PHYSIO POV.pptxNEONATAL RESPIRATORY CARE FROM A PHYSIO POV.pptx
NEONATAL RESPIRATORY CARE FROM A PHYSIO POV.pptx
 
Understanding Warts and Moles: Differences, Types, and Common Locations
Understanding Warts and Moles: Differences, Types, and Common LocationsUnderstanding Warts and Moles: Differences, Types, and Common Locations
Understanding Warts and Moles: Differences, Types, and Common Locations
 
ARTHRITIS.pptx Prepared by monika gopal Tutor
ARTHRITIS.pptx Prepared  by monika gopal TutorARTHRITIS.pptx Prepared  by monika gopal Tutor
ARTHRITIS.pptx Prepared by monika gopal Tutor
 
Three Keys to a Successful Margin: Charges, Costs, and Labor
Three Keys to a Successful Margin: Charges, Costs, and LaborThree Keys to a Successful Margin: Charges, Costs, and Labor
Three Keys to a Successful Margin: Charges, Costs, and Labor
 
Artificial Intelligence: Diabetes Management
Artificial Intelligence: Diabetes ManagementArtificial Intelligence: Diabetes Management
Artificial Intelligence: Diabetes Management
 
Basics of Giant Cell Tumor of bone (GCTB)
Basics of Giant Cell Tumor of bone (GCTB)Basics of Giant Cell Tumor of bone (GCTB)
Basics of Giant Cell Tumor of bone (GCTB)
 
Painting Rats White Angers Them to No End
Painting Rats White Angers Them to No EndPainting Rats White Angers Them to No End
Painting Rats White Angers Them to No End
 
LARYNGEAL CANCER.pptx Prepared by Neha Kewat
LARYNGEAL CANCER.pptx  Prepared by Neha KewatLARYNGEAL CANCER.pptx  Prepared by Neha Kewat
LARYNGEAL CANCER.pptx Prepared by Neha Kewat
 
Living Well Every Day: Lyons Wellness Practice | Nurtures Your Complete Health
Living Well Every Day: Lyons Wellness Practice | Nurtures Your Complete HealthLiving Well Every Day: Lyons Wellness Practice | Nurtures Your Complete Health
Living Well Every Day: Lyons Wellness Practice | Nurtures Your Complete Health
 
Diseases of the Respiratory System (J00-J99),.pptx
Diseases of the Respiratory System (J00-J99),.pptxDiseases of the Respiratory System (J00-J99),.pptx
Diseases of the Respiratory System (J00-J99),.pptx
 
The Power of Active listening - Tool in effective communication.pdf
The Power of Active listening - Tool in effective communication.pdfThe Power of Active listening - Tool in effective communication.pdf
The Power of Active listening - Tool in effective communication.pdf
 

Semen analysis by Dr.Renukadevi

  • 1. SEMEN ANALYSIS DR.RENUKADEVI SENIOR EMBRYOLOGIST ARC International Fertility & Research Centre
  • 2. Introduction  A semen analysis measures the amount of semen a man produces and determines the number and quality of sperm in the semen sample.  A semen analysis is usually on e of the first test done to help determine whether a man has problem fathering a child (infertility)  A problem with the semen or sperm affects more than one- third of the couples who are unable to have children (infertile)
  • 3. Semen Analysis  SA are mainly carried out to determine whether:-  A man has a reproductive problem that is causing infertility.  A vasectomy or vasectomy reversal has been successful  SA is the singular test for fertility in male that can provide information on ◦ sperm production. ◦ patency of the male ducts. ◦ the function of the accessory glands. ◦ and ejaculative function. Since semen samples may vary from day to day, 2 or 3 samples may be evaluated within a 3-6 month period for accurate testing. A semen analysis to test the effectiveness of a vasectomy is usually done 6 weeks after the vasectomy
  • 4. Semen collection  Masturbation, directing the semen into a clean sample cup. Do not use a lubricant which can kill sperms  Coitus interruptus - withdrawing the penis from the partner just before ejaculating follow by ejaculating into a clean sample cup.  Coitus - by using a condom. A special (silicon) condom that does not contain any substance that kills sperm (spermicide). After ejaculation, carefully remove the condom from the penis. Tie a knot in the open end of the condom and place it in a container that can be sealed in case the condom leaks or breaks. Ordinary condoms should not be used since they usually contain spermicides  Assisted ejaculation – electro-ejaculation used in paralegics
  • 5.  Once after collecting the semen sample from the patient, receiving time is noted down.  The sample has to be liquefied for 20-30mins at 37°c by placing the sample in heating block. (Avoid crystallization by leaving the sample stationary for a too long time).  After 20-30mins of liquefaction, mix the sample homogeneously by using transfer pipette.  Sample volume, viscosity and pH should be noted. SEMEN ANALYSIS
  • 6. Semen Collection • Abstinence days 2-5 • Pass urine • Wash hands with soap, dry • Collect the entire sample into the wide mouth sterile container, 70% of sperms is in the first part of the ejaculate • Keep the sample at body temperature, no sunlight • Deliver the sample within one hour of ejaculation Good and reliable SA results starts from semen collection, preferably by masturbation
  • 7. Parameters (WHO criteria 2010) Parameters agreed on consensus Parameter Lower Reference Limit Semen volume (ml) 1.5 Sperm concentration (106/ml) 15 Total sperm number (106/ejaculate) 39 Progressive motility (PR, %) 32 Total motility (PR +NP, %) 40 Vitality (live sperms, %) 58 Sperm morphology (NF, %) 4 pH* >/=7.2 Leucocyte* (106/ml) <1 MAR/Immunobead test* (%) <50
  • 8. Terminologies of Semen Analysis  Oligospermia – sperm concentration <15 million/ml  Asthenozoospermia – <40% grade (PR+NP) or < 32 PR%  Teratozoospermia – <4% spermatozoa  OAT =Oligo-astheno-teratozoospermia  Azoospermia – no spermatozoa in semen  Polyzoospermia – ++ high sperm concentration, >200M/ml  Hypospermia – semen volume < 1.5 ml  Hyperspermia – semen volume > 4.5 ml  Aspermia – no semen volume  Pyospermia – leukocytes present in semen, >1M/ml  Hematospermia – red blood cell present in semen  Necrozoospermia – “dead” sperm
  • 9. Macroscopic Examination WHO criteria 2010 Description Appearance Normal: Whitish to grey opalescent Yellow (urine, jaundice); Pink/Reddish/Brown (RBCs) Liquefaction Normal: 15–30 minutes after collection Lumpy >60 min – sperms may be trapped in unliquefied jelly; maybe sign of prostatic infection, lack of prostatic protease Viscosity Normal Smooth and watery Abnormal –, thick with long threads (21G needle). High viscosity impede sperm movements MIX SEMEN THOROUGHLY BEFORE ANALYSIS –USE A PIPETTE
  • 10. Macroscopic Examination Volume Normal: 1.5 ml per ejaculation Low volume (<1ml) reflect a problem with the seminal vesicles and prostate – a block, retrograde ejaculation, infection or lack of androgen. Low semen volume cannot neutralize vaginal acidity High semen volume dilute sperms/ active infection pH Normal: =/>7.2 (alkaline) Acidic pH (<7.0) in a low volume & density sample indicates –congenital bilateral absence of vas deferens (in which seminal vesicles are also poorly developed) and ejaculatory duct obstruction. pH increases with time as natural buffering capacity of semen decreases – therefore high ph is not clinically useful
  • 14.  A microscope objective of 20x should be used for counting.  Place a drop of the sample into Makler chamber and count the sperm heads as follows: 10 squares of the field. This number represents the concentration of spermatozoa in millions per milliliter.  When counting a sample of oligozoospermia (less than 15million/ml) all 100 squares have to be counted. Adding "00000" to that number, you will get the concentration of the spermatozoa per milliliter. SPERM COUNT Microscopic Examination
  • 15.  Grade A (fast progressive) sperms are those which swim forward fast in a straight line - like guided missiles.  Grade B (slow progressive) sperms swim forward, but either in a curved or crooked line, or slowly (slow linear or non linear motility).  Grade C (non-progressive) sperms move their tails, but do not move forward (local motility only).  Grade D (immotile ) sperms do not move at all.  Immotile spermatozoa are to be counted in a defined number of squares. Afterwards the motile spermatozoa are counted and classified into four categories (from A "fast progressive" to D "no motility"). This procedure is to be repeated in different areas of the grid to have the quality and motility to be calculated on base of this data in percent. Motility is graded from A to D,
  • 20. SPERM MORPHOLOGY A drop of sample is placed in the sterile glass slide and covered using a cover slip, name and SID number should be mentioned. The sample is viewed under light microscope with a magnification of 40x. A Normal Sperm has: A smooth, oval-shaped head that is 5-6 micrometers long and 2.5-3.5 micrometers wide (less than the size of a needle point) A well-defined cap (acrosome) that covers 40% to 60% of the sperm head No visible abnormality of neck, mid-piece, or tail No fluid droplets in the sperm head that are bigger than one half of the sperm head size.
  • 22. The following categories of defects should be noted. Head defects, namely large, small, tapered, pyriform, globe, amorphous heads, vacuolated heads, double heads and small acrosome. Neck and mid-piece defects namely bent neck, abnormal mid-piece (thin or thick). Tail defects namely short, multiple, hairpin, broken, bent tails, coiled tail. Multiple defects Agglutination Pus cells/HPF Debris After completion of analysis the results should be noted in the record and typed in the patient report entry column.
  • 23. Morphology assessment  Sperms must be stained for an accurate assessment  Strict criteria /Kruger /Tygerberg  The sperm head is oval, smooth-symmetrical outline, a length of 3-5 um and a width of 2-3um.  The head should have a well defined acrosome area of 40-70% and vacuoles (=/<2) that occupies ~ 20% head area.  The mid-piece must be straight and slender, 0.5 um in width and 7-8um long, straightly aligned to the head.  The tail must be straight and 45-50 um long.  To be classified as normal, the sperm must be normal in all portions (head, mid-piece, tail).  At least 400 sperms must be scored on randomly chosen fields. Normal Forms (%) = normal sperms / the total number of sperms evaluated x 100.
  • 24. Diff-Quick Staining  Simple, quick staining method.  Moderate differentiation of structures --the acrosome is stained red and the post acrosomal area dark red. Very thin smears are necessary as the background is influenced by the presence of protein in the seminal fluid and tends to be dark.  The slides are fixed with fixative (Cytospray, Kinetik, Australia).  Diff Quik 1 for 5-6 seconds.  Clean water x 2. Removed excess moisture.  Diff Quik 2 for 10 seconds  2-3 dips in clean water.  It is left to dry by standing the slide on one end on absorbent paper towel.  Sperms stained by Diff-Quik are larger by comparison to H&E staining as the cells did not undergo dehydration by alcohol.
  • 26. Abnormal Sperms 1. Triple head sperm 2. Acrosome reacted sperm 3. Sperm with no acrosome 4. Sperm with a tapering head and swollen mid-piece
  • 27. Morphology Assessment  Assessment is subjective due to the wide variation in sperm sizes and shapes.  % NF correlated well with the fertilization rate in-vitro and pregnancy rate [Kruger et al, 1996; Morgenthaler et al, 1995]  Sperms with defective heads are more likely to be immotile than sperms without defects, and defective motile sperms tend to show sluggish motility as compared to normal sperms [Aitken et al, 1995].
  • 28. Vitality Assessment 1. Eosin-nigrosin (dead sperm stain pink/red) 2. Eosin (1%) (dead sperm stain pink/red) 3. Trypan (0.4%) blue (dead sperm stain blue) 4. Hypo-osmotic swelling test (HOS) (live sperm shows tail curling Test 1, 2 and 3 for diagnostic uses.  Usually 1:1 ratio of semen to dye mixture, mix well and smear onto a slide. Read immediately at x40 objective, count 200 sperms Test 4 is use to choose live (immotile) sperm for ICSI  Dead sperms will not react in HOS while live sperm will take up fluids causing their tails to curl within 5 min and stabilize at 30 min. Therefore viable sperms may be selected for ICSI [Lin et al, 1998; Cayan et al, 2001].
  • 29. Vitality Assessment Dead sperms are stained pink/red Live sperms shows curling tails