VI
TALI
TYSTAI
N
TAKEAGLASSSLI
DE&CLEANI
TBOTHTHESI
DES
MAKETHESEMENSAMPLEREADYFROM THESAMPLEWARMERAT
37DERGREECELCI
OUS
ADDSEMENSAMPLEABOUT50MI
CROLI
TTERI
NTHE1MLTUBE&
ADD50MI
CROLI
TTERVI
TALI
TYSTAI
N
MI
XWELL&I
NCUBATEI
TFOR2MI
NI
TUES
TAKE10MI
CROLI
TTEROFTHI
SMI
XTURE
PLACEI
TI
NTHEGLASSSLI
DEONTHERI
GHTSI
DE&MAKEATHI
CK
SMEARE
DRYI
TI
NTHESLI
DEWARMER
OBSERVEI
TI
NTHEMI
CROSCOPEAT40XOBECTI
VE.
MORPHOLOGYSTAI
N
CENTRI
FUGETHESEMENSAMPLEFOR3MI
NI
TUESAT1500RPM &
REMOVETHESUPERNATANT
ADDWASHMEDI
UM I
NTHEPELET&CENTRI
FUEI
TAGAI
NI
NTHE
SAMERPM(REPETTHI
SFOR3TI
MES)
THENREMOVETHESUPERNATANTAGAI
N&ADDMI
NI
MUM AMOUNT
OFWASHMI
DI
UM
THENMI
XWELL&TAKE10MI
CROLI
TTEROFTHI
SSAMPLE
PLACEI
TI
NCLEANGLASSSLI
DE&MAKEATHI
KSMEAR
DRYI
TI
NSLI
DEWARMER&PLACEI
TI
NTHESTAI
NI
NGTRAY
ADDFI
XATI
VE1ANDWAI
TFOR15SECONDS&DRAI
NI
T
ADDSTAI
N1WAI
TFOR10SECONDS&DRAI
NI
T
ADDSTAI
N3WAI
TFOR9SECONDS&I
MMI
DI
ATELYWASHI
TWI
TH
DI
STI
LLWATER
DRYI
TWI
THSLI
DEWARMERUNTI
LDRY
OBSERVEI
TI
NMI
CROSCOPEAT100XOI
LEMERSSI
ON&COUNT100
SPERMS
DNASTAI
N
PLACETHEGELLI
NTHEDNAWARMERAT75ºFOR20MI
NI
TUES
AFTERMELTI
NGTHEGELLPLACEI
TAT37º
CSAMPLEWARMER
FOR5MI
NI
TUES
ADD60µl
OFWASHEDSAMPLEI
NTHEGELLANDMI
XWELL
WARM I
TFOR5MI
NI
TUESI
N37º
C
TAKEADNASLI
DE&PLACE15µl
OFTHEGELLSAMPLEI
NTHE
CENTERPARTOFTHESLI
DE&PLACETHECOVERSLI
PWI
THOUT
BUBBLE
KEEPTHESLI
DEAT2º-4ºFOR5MI
NI
TUES.
THENREMOVETHE
COVERSLI
PGENTLY
PLACETHI
SSLI
DEI
NTHEI
MMERSI
ONTRAY&ADD3MLOFLYSE1&
KEEPI
TFOR7MI
NI
TUES.
TAKETHESLI
DEANDPLACEI
TI
NTHESECONDPOSSI
TI
ONOFTHE
SAMETRAY&ADD3MLOFLYSE2.KEEPI
TFOR25MI
NI
TUES.
TAKE&CLEANTHEBACKSI
DEOFTHESLI
DEPLACEI
TI
NTHE3rd
POSSI
TI
ON &ADD5MLDI
STI
LLEDWATTER,
KEEPI
TFOR5MI
NI
TUES
THENPLACETHESLI
DEI
NTHETRAY&COVERI
TWI
THFI
XATI
VE1
FOR2MI
NI
TUES
DRAI
NI
T &ADDFI
XATI
VE2FOR2MI
NI
TUESFOLLOWEDBYFI
XATI
VE3FOR2MI
NI
TUES&DRAI
NI
T.
ADDSTAI
N1,
1MLI
NCUBATEFOR2MI
NUTES.ADDBUFFER2ML
WI
THOUTDRAI
NTHESTAI
N–1&KEEPI
TFOR15MI
NI
TUES
RI
NCEI
TWI
THDI
STI
LLEDWATER&DRYI
T.OBSERVEI
TI
N40X
OBJECTI
VE
COUNTMI
NI
MUM 100-300SEMENPERSLI
DES.
FORANYCLARI
FI
CATI
ONPLCALL
BORUS
TECHNI
CAL&MARKETI
NGMANAGER
9940046139