HANDLING OF FROZEN SEMEN

3s be the maximum time for moving 0.25 ml

straws from one liquid nitrogen tank to another
without damaging the sperm and similarly for
moving from tank to thaw flask

General Within the liquid nitrogen storage tank

Recommendation have the canisters containing the
straws clearly labelled.

Maintain an up-to-date inventory of

semen straws, bull codes and their
location within the liquid nitrogen tank

When locating straws keep the canister

below the frost line and avoid lifting the
canister for too long in the process.
Management of Cryo container

Properties of LN

• Extreme low temperatures.

• Large ratio of expansion in volume
from liquid to gas.
• Most cryogenic liquids are odourless
and colourless when vapourised to gas.
• Boiling point: -195.8 0 C
• Storage : The most commonly used
container for handling cryogenic fluids is
the Dewar flask.
• Dewar flasks are non pressurized, vacuum
jacketed vessels.
Parts of Cryocontainer
FRP Neck
Handle
Lock cover
Vaccum Nozzle
Neck Plug
Adsorbent
Cannister
Inner liner
Shell
 Management of Cryocontainer

Dos and DONTs handling L N

Container
 Care and management of the LN Container

Avoid direct DONOT Tilt Avoid Always keep Always keep Tipping the
contact with or roll the welding/ the the container container or
hard base. Keep LN drilling in container vertically in a letting it lie
the LN Container to the closed with well may damage
Container in jute avoid container the Neck ventilated the container
carpet or splashing wall plug to area and the
wooden plank minimize materials
LN2 loss. inside
Use funnel to Use wodden or In case of Condensed Use forceps or
avoid metal dipstick asphyxiation ! moisture or straw to
splashing and to measure Immediately frost on the remove cane
spilling while the LN Level contact doctor outside are from the
transferring indications of storage
LN2 vaccum loss container
 Measuring LN 2 Level

Measure the
level of liquid
Open the nitrogen 1
The Remove cm below the
evaporation lid, pass
the stick the stick level of frost
rate of and wave line
liquid into the
in the air •A minimum
nitrogen To measure container
till it for few of 15 cm
varies the level of seconds should be
between the LN2, the reaches the
bottom of which will maintained
containers. dipstick is leads to when the
Hence the the
used frost straws are
periodic container.
Keep the formation stored in the
checking of container
the level of stick for 10
liquid seconds
nitrogen is
essential.
 Proceed with Thawing of semen
preparation of gun
only when sure of straw (as per SOP)
heat

Remove the semen straw from the

Have plastic gloves, Add water at 35 to container with precooled forceps
sheath, gun, scissors, 37 degree and not with hands. While taking
forceps, tissue paper, centigrade to the out, raise the canister – not above
and clean towel ready thawing tray. the frost line
before thawing semen

Frost line
• Remove the straw from the water bath, wipe dry, Load the semen straw onto
the gun and cut the straw at a right angle with a straight and sharp scissors
just below the laboratory seal.

• Take out the sheath by holding bottom of the sheath through a small hole at
the corner of the sheath packet and place the sheath on the gun and secure
the sheath firmly with o-ring lock. To ensure better hygiene, use individually
packed sheaths instead of sheaths available in bulk packing.

• If there is a delay (>10 to 15 s) before insemination wrap the loaded gun in in a

towel to provide hygienic and thermal protection or place in a gun warmer.

• In very warm environments it is important that loaded AI guns are protected

from solar radiation and elevation and temperatures above 35°C.
• Clean the vulva of the cow with paper towel before insemination.