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W evaIuates certaIn

cbaracterIstIcs oI a maIe's
semen and tbe sperm
contaIned In tbe
W measures tbe amount oI
semen a man produces.
W determInes tbe number and
quaIIty oI sperm In tbe
semen sampIe.
W aIso caIIed sem1no1 ]1u1d presen1 1n
W medIum tbe sperm can swIm
W contaIns spermatozoa
W a vIscId mucIIagInous IIuId wItb IaInt
yeIIow coIor
W Has cbaracterIstIc odor caIIed semInaI
$emen consIst oI tbe IoIIowIng
1. $permatozoa (10%)
2. $emInaI PIasma (90%)
3. EpItbeIIaI eII (< 1%)
W $perm
-reIers to tbe
maIe reproductIve
Purpose oI $emen AnaIysIs
1. To verIIy tbe success oI vasectomy
(usuaIIy done 6 weeks aIter tbe
Vasectomy- a surgIcaI procedure In
wbIcb tbe vasa deIerentIa oI a man
are severed, and sometImes tIed or
seaIed. TbIs resuIts to sterIIIty.
2. to InvestIgate InIertIIIty
PrerequIsItes Ior semen
W AvoId Iong perIod oI sexuaI
W AvoId any sexuaI actIvIty tbat
resuIts In ejacuIatIon Ior 2 to S
days beIore semen coIIectIon.
W Tbe subject sbouId be provIded wItb
cIearIy wrItten or oraI InstructIons
concernIng tbe coIIectIon and
transport oI tbe semen sampIe.
W Tbe name oI tbe person, perIod oI
abstInence, date and tIme oI
coIIectIon, and any medIcInes or
berbaI suppIements tbat tbe person
Is takIng sbouId be recorded.
$ampIe coIIectIon and
1. Iean tbe bands and penIs
2. Tbe sampIe sbouId be obtaIned by
ays oI coIIectIon
W MasturbatIon
- a sexuaI stImuIatIon (not
Intercourse) tbat produces an orgasm.
W $urgIcaI Procedure
W DurIng Intercourse usIng a non-
spermIcIdaI condom
3 types oI surgIcaI
MIcrosurgIcaI EpIdIdymaI
$perm AspIratIon (ME$A)
-used In patIents wItb
reproductIve tract bIockage.
TestIcuIar $perm
AspIratIon (TE$A)
- tbe testIcuIar tIssue Is sucked
out tbrougb a IIne needIe, under
IocaI anaestbesIa.
Percutaneous EpIdIdymaI $perm
AspIratIon (PE$A)
tbe sperm Is sucked out Irom tbe
epIdIdymIs by puncturIng It wItb a
IIne needIe.
W 411us 1n1errup1us aIso
known as wItbdrawaI or tbe
puII-out metbod. It may be
used aIter a vasectomy to test
Ior tbe presence oI sperm, but
It Is not acceptabIe as a means
oI coIIectIon because oI tbe
W tbe IIrst portIon oI tbe ejacuIate,
wbIcb contaIns tbe bIgbest
concentratIon oI spermatozoa, may
be Iost
W tbere wIII be ceIIuIar and
bacterIoIogIcaI contamInatIon oI
tbe sampIe
W tbe acId pH oI tbe vagInaI IIuId wIII
adverseIy aIIect sperm motIIIty
3. Tbe sperm Is ejacuIated Into a
cIean, wIde-moutbed gIass or
pIastIc contaIner, or a non-
spermIcIdaI condom.
ontaIner Ior $emen sampIe
W Iean, wIde-moutbed gIass
W Iean, wIde-moutbed pIastIc (no
toxIc eIIect on spermatozoa)
W Non-spermIcIdaI condom (not just
tbe ordInary condoms)
*contaIners sbouId be warm to
mInImIze tbe rIsk oI coId sbock
. It Is recommended to get tbe IIrst
part oI tbe ejacuIatIon as It Is
tbougbt to be tbe most sperm-rIcb.
S. Two semen sampIes sbouId be
coIIected Ior InItIaI evaIuatIon.
AddItIonaI semen sampIes may be
6. Protect tbe sampIe Irom extremes
oI temperature (not Iess tban 20c
and not more tban 0c) durIng
transport to tbe Iaboratory.
?. Tbe sampIe sbouId be examIned
ImmedIateIy aIter IIqueIactIon and
wItbIn 1b oI ejacuIatIon
#uIes In semen coIIectIon and
W Tbe sampIe sbouId be coIIected
aIter a mInImum oI bours and no
Ionger tban ? days oI sexuaI
* DuratIon oI abstInence
W more tban ? day: sperm motIIIty
may decIIne
W <b: sperm concentratIon may
decIIne, but tbe same motIIIty
W Tbe tIme IntervaI between tbe Iast
ejacuIatIon and sampIe coIIectIon
sbouId be weII deIIned and
preIerentIaIIy as constant as possIbIe.
W Tbe name oI tbe person, perIod oI
abstInence, date and tIme oI coIIectIon
sbouId be recorded.
W Any medIcInes or berbaI
suppIements tbat tbe person Is
takIng sbouId be noted
W Two semen sampIes sbouId be
coIIected Ior InItIaI evaIuatIon.
W Tbe coIIected sampIes sbouId not
be Iess tban ? days or more tban 3
montbs apart.
W Tbe bands and penIs sbouId be
cIean wben masturbatIng.
W Tbe sampIe must be deIIvered to
tbe Iaboratory wItbIn 1 bour aIter
W $emen oIder tban 1 bour sbouId not
be accepted Ior .rg4preservo114n
(rapId IreezIng In IIquId or vapor
nItrogen at -196 to preserve
sperm Ior Iuture use).
W Tbe sampIe sbouId be stored In
temperature not Iess tban 20c and
not more tban 0c durIng
transport to tbe Iaboratory.
W Do not reIrIgerate tbe sampIes.
W HandIe tbe sampIe wItb care.
&nacceptabIe $pecImens
W a. contaIner Is cracked, broken or
W b. contaIner not approved by tbe
W c. contaIner Is bot or coId to tbe
W d. specImen coIIected by use oI
condom not approved by tbe
W e. specImen brougbt In by
unscbeduIed waIk-In appoIntments.
W I. IncompIete sampIes, wbereIn tbe
IIrst portIon oI tbe ejacuIate Is Iost.
$emen ExamInatIon
W Tbe Parameters In TestIng can be
cIassIIIed In two categorIes:
MacroscopIc ExamInatIon and
bemIcaI ExamInatIon
W $perm ount
W VoIume
W MorpboIogy
W MotIIIty
W oIor
W VIscosIty
W LIqueIIcatIon tIme
TotaI sperm count
W %41o1 sperm .4un1, or 141o1 sperm
number, Is tbe totaI number oI
spermatozoa In tbe entIre ejacuIate.
By H, Iower reIerence IImIt Is 39
mIIIIon per ejacuIate
W Average sperm concentratIon Is
more tban 60 mIIIIon per mIIIIIIter
(>60 mIIIIonJcc). ounts oI Iess
tban 20 mIIIIon per mIIIIIIter (<20
mIIIIonJcc) are consIdered sub-
NormaI ejacuIated voIume Is between 2
and 6 mI.
H regards 1.S mI as tbe Iower
reIerence IImIt
Low voIume may IndIcate partIaI or
compIete bIockage oI tbe semInaI
vesIcIes or tbat tbe man was born
wItbout semInaI vesIcIes.
W IassIIIcatIons:
NormaI ovaI-sbaped
NormaI spermatozoId
vaI Iorm wItb smootb contour
AcrosomaI cap encompassIng 0-?0%
oI bead
No abnormaIItIes oI mIdpIece or taII
No cytopIasmIc vacuoIes oI more tban
baII oI tbe sperm bead.
Head sIze oI S-6 M x 2.S-3.S M
$trIct crIterIa Ior normaI
sperm morpboIogy:
W $perm bead
$mootb ovaI conIIguratIon
Lengtb: S-6 mIcrons
Idtb: 2.S-3.S mIcrons
Acrosome comprIses 0-?0% oI tbe
anterIor sperm bead.
W MIdpIece
AxIaIIy attacbed
, 1.S tImes tbe bead Iengtb
Iess tban or equaI to 1 m In wIdtb
sIIgbtIy tbInner tban tbe mIdpIece
S m Iong
$perm morpboIogy
$taInIng metbods
Tbe PapanIcoIaou staIn and tbe
$borr staIn are tbe metbod most
wIdeIy used
Tbe DIII-QuIk rapId staInIng metbod
Is used too
W It may bave background staInIng
and may not aIways gIve tbe same
quaIIty as tbe PapanIcoIaou staIn or
$borr staIn.
$perm morpboIogy
PIoios and scIcmaiic drawings of somc ,-3472,
14728of Iuman scrmaiozoa.
NormaI sperm bead

Absence oI the acrosome and

pointed anterior head segment
Very small acrosomal area
AbnormaI sperm bead

$mall size and absence oI acrosome with

typical round-head deIect or globozoospermia
Moderate to severe elongation
oI the postacrosomal area
AbnormaI sperm bead
Sperm Pead
W sperm1o: No semen ejacuIated
W emo14sperm1o: BIood present In
W eu.4.g14sperm1o: bIte bIood
ceIIs present In semen
W 44sperm1o: No spermatozoa
Iound In semen
W 11g444sperm1o: Low sperm
W s1en444sperm1o: Poor motIIIty
andJor Iorward progressIon
W %ero1444sperm1o: #educed
percentage oI morpboIogIcaI
normaI sperm
W e.r444sperm1o: No IIve sperm In
W 14b444sperm1o: #ound beaded
acrosome-Iess sperm
W TbIs descrIbes tbe percentage oI
sperm tbat
are movIng. FIIty percent or more oI
tbe sperm sbouId be movIng.
W Tbe H deIInes normaI motIIIty 60%
oI observed sperm or at Ieast mIIIIon
per mIIIIIItre, sbowIng good Iorward
tbIs must be measured wItbIn 60
oI coIIectIon.
W H aIso bas a parameter oI
wItb a Iower reIerence IImIt oI
60% IIve spermatozoa.
W TotaI MotIIe ount:
TbIs Is tbe number oI movIng sperm
In tbe entIre ejacuIate.
W It Is caIcuIated by muItIpIyIng tbe
voIume (cc) by tbe concentratIon
(mIIIIon spermJcc) by tbe motIIIty
(% movIng). Tbere sbouId be more
tban 0 mIIIIon motIIe sperm In tbe
Tbe Iour-IeveI grade system
0 No movement
,ovement none forward
+ Cccas|ona| movement of a few sperm
S|ow und|rected
S|ow d|rect|y forward movement
Iast but und|rected movement
Iast d|rected forward movement
+ Iery fasL frward memeL
4 Lxtreme|y fast forward movement
W rode 4 sperm are known to bave
progressIve motIIIty meanIng tbey
are tbe strongest and swIm Iast In a
straIgbt IIne.
W rode S sperm (non-IInear motIIIty)
aIso move Iorward but tend to
traveI In a curved or crooked
W rode 2 sperm are IabeIIed as non-
progressIve motIIIty because tbey
do not move Iorward despIte tbe
Iact tbat tbey move tbeIr taIIs.
W rode J sperm are ImmotIIe
meanIng tbat tbey IaII to move at
W IdeaI sperm quaIIty dIctates tbat a
man sbouId bave grade 3 or
sperm In order to IertIIIze an egg.
W NormaIIy, >S0% oI sperm In tbe
specImen are motIIe.
W Forward progressIon descrIbes bow
Iast tbe motIIe sperm are movIng
(normaI 2+ In tbe scaIe Irom 0 to )
$tandard $emen FIuId Tests
W 4mp4sed 4] 1e ono1gs1s 4] 414r,
v1s.4s11g (bow tbIck tbe semen Is),
and tbe 11me un111 1e spe.1men
W oIor and TurbIdIty
$emen Is normaIIy transIucent or
wbItIsb-gray opaIescent In coIor.
W BIood Iound In semen
(bematospermIa) can coIor tbe
semen pInk to brIgbt red to
brownIsb red. Tbe presence oI bIood
In semen Is abnormaI and sbouId be
reported. Tbe presence oI partIcIes,
nonIIquIIIed streaks oI mucus or
debrIs requIres Iurtber evaIuatIon.
LIqueIIcatIon TIme
W $emen Is normaIIy produced as a
coaguIum. Tbe specImen wIII
ususaIIy IIquIIy wItbIn 30 mInutes.
W Tbe IaIIure to IIquIIy wItbIn one
bour Is abnormaI.
VIscosIty Is measured aIter
compIete IIqueIactIon bas occured.
VIscosIty Is consIdered "normaI" II
tbe IIqueIIed specImen can be
poured Irom a graduated beaker
drop by drop wItb no attacbIng
aggIutInum between drops.
W Hyper -vIscosIty may InterIere wItb
tbe abIIIty oI sperm to traveI Irom
tbe sIte oI deposItIon Into tbe
cervIx or uterus.
H crIterIa specIIy normaI as ?.2-
AcIdIc ejacuIate (Iower pH vaIue) may
IndIcate one or botb oI tbe semInaI
vesIcIes are bIocked.
A basIc ejacuIate (bIgber pH vaIue)
may IndIcate an InIectIon
W W procedure tbat determIne Iructose procedure tbat determIne Iructose
content In semen. content In semen.
W W InvoIves beatIng semen In strong InvoIves beatIng semen In strong
acId acId
wItb tbe presence oI resorcInoI. wItb tbe presence oI resorcInoI.
+ #E$&LT: a red coIor In soIutIon + #E$&LT: a red coIor In soIutIon
A PosItIve resuIt In Fructose
Test Is normaI.
A negatIve resuIt Iructose may:
1. IndIcate bIocked semInaI vesIcIes
2. Tbe $perm Iacks an energy source
LeadIng to maIe InIertIIIty
MA#(mIxed antIgIobuIIn
reactIon ) test
ExamInatIon oI ImmunoIogIcaI
InIertIIIty (anaIysIs oI sperm
membrane antIbodIes)
NormaI vaIue: Iewer tban 10%
spermatozoa wItb adberent
EosIne-nIgrosIne test
W Tbe proportIon oI IIve spermatozoa can
be determIned by usIng staInIng
tecbnIques tbat are
W based on tbe prIncIpIe tbat dead ceIIs
wItb a damaged pIasma membrane take up
certaIn staIns
H$ test - Hypo smotIc
$weIIIng Test
W test based on tbe semI-
permeabIIIty oI tbe
Intact ceII membrane wben an
InIIux oI
water resuIts In an expansIon oI ceII
$perm bromatIn $tructure
Assay ($$A).
W test Ior sperm DNA IragmentatIon
W sperm are wItb a AcrIdIne orange
W (coIored dye onIy attacbes to
broken DNA )
W II tbe DNA Is Intact tben no dye wIII
attacb to tbe sperm.
W A IIow cytometer Is used to anaIyze
ten tbousand sperm Irom tbe
W A computer soItware aIIows
caIcuIatIng tbe DNA IragmentatIon
Index (DFI).
W LLp//wwwaDFI &suaI vaIues :
Very good : DFI : < 1S %
Bad : DFI : > 30 %
W laascm/rmal_seme_aluesLml