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Artificial insemination thawing Storage Freezing the equilibration Packaging Glycerolation? Dilution Sperm collection and examination (CRYOPRESERVATİON)

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(1)

(CRYOPRESERVATİON)

Sperm collection and examination

Dilution

Glycerolation?

Packaging

the equilibration

Freezing

Storage

thawing

Artificial insemination

(2)

Sulandırma

Dilution (ratio, temp, pH, osmotic press)

Tris

Tes

Pipes

Bes

Commercial (lasifos, biosifos)

external cryoprotektanlar

Egg yolk (%10-20)

BSA (3-6 mg/ml)

Soya proteini

İnternal cryoprotektanlar

DMSO

DMA (%1-15)

Gliserol

Etilen glikol

Antioksidanlar

Vit e, a

Taurin

Heparin

Hipotaurin

Glikozaminoglikanla r

Antibiyotikler

Gentamisin, penisilin-

streptomisin

(3)
(4)

Gliserolizasyon

 Sulandırıcının gliserollü kısmı ile sulandırma

 Gliserol (%1-15)

 Boğa için %7

(5)

PAKETLEME

 PELLET (0.1 ML)

 MİNİ PAYET (0.25, 0.50 ML)

 MAKROPAYET (2-5 ML)

 YASSI PAYET (2.5 ML)

 AMPUL

 VİAL

 TÜP

(6)

Ekilibrasyon (alışım)

 4

o

C de 5 dakika ile 4 saat arası

 boğa için 2 saat

Amaç:

Spermatozoonun sulandırıcıya alışması Sıcaklığın düşürülmesi

Cryoprotektana alışım

(7)

(cryopreservation)

PELLET

 İce (KATI CO

2

, -79

o

C) PAYET

 Liquid nitrogen vapor (-110-120

o

C

 Automatic freezing machine

Protocols

Dry ice 5 min (-79

o

C)

Liquid nitrogen vapor 10 min (3 cm above liq, -120

o

C )

(8)
(9)
(10)

Storage

Referanslar

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