Collect semen in a sterile specimen cup, let liquefy ( could take up to 15 minutes) this is when the sperm turns from a thick gel-like consistency to a liquid.
Swirl in the cup if it is still thick after this time or pipette repeatedly till homogeneous.
Step 2: Warm TYB during liquefaction at 98.6F or 37C in a dry cell or water bath. Make sure to dry outside of the vial if using warm water.
Step 3: Uptake 5ml of TYB with a sterile syringe
Step 4: Add drop by drop with a 18G to 24G needle at a 1:1 ratio ( example 1ml semen to 1ml TYB) should be done slowly so not to form a stream of TYB. Not Less than 2 minutes per 3ml of TYB as not to cause osmotic shock to the sperm cells.
Step 5: place specimen vials between two gel packs room temp 60-70F
step 6: cool down in a fridge at 41F to 50F make sure to keep away from the back of the fridge as the compressor will FREEZE AND KILL THE SPERM CELLS. DO NOT FREEZE
Keep close to the door or veggie drawer on the bottom of the fridge.
Remove at 24hrs or when you plan to test the specimen under a microscope. Warm at body temp for 10-15 minutes to see motility.
Step 1 Make sure your 3ml cryo vial with TYB and sperm at 1:1 ratio are SEALED TIGHTLY.
Step 2 take warm water between room temp and 98.6( most ideal) and warm the specimen for 10 to 15 minutes.
Step 2 alternative ( warm specimen vials in waistband or against body for 10-15 minutes)
Step 3 Dry the outside of the vial thoroughly if you used a water bath!! Now begin to rock the vial back and forth to resuspend the settled sperm cells.
Step 4 continue rocking cells for 1 minute for 20 tilts a minute.
Step 5 remove your pipette and start sucking the liquid from the bottom of the vial and stop uptake upon the remainder of the liquid being filled into the pipette. This will prevent air bubbles from forming
Step 6 take your pipette and place one drop on a microscope slide and cover it with a glass coverslip. View at 400X under a light microscope to view motility.
You may now Use or discard the remainder of the semen TYB mixture for further scientific-educational observation. Take notes, see how fast they swim!
With a hemocytometer, you may count the viable cells and dead cells.
After cooling in TYB for a few hours 2-3) You can also ship your specimen to a LAB or someone that does CASA ( computer-aided semen analysis ) if you are curious in having your test performed by a professional. one such service that does this is https://www.dadikit.com/ ( not affiliated ) they provide a video of your sperm motility and total sperm count + morphology results.
Step 1: After mixing Either pre-cool specimen at 41F for 2-3 hours ( not necessary ) or place the sperm vials directly on 41F gel packs and place in a foam cooler to ship 24hr overnight. vials will cool naturally. If it is summer place a frozen gel pack OUTSIDE of the foam cooler to help keep it at temp and wrap in a mylar blanket. followed by your outside UPS or fedex package.
See the pictures below for more details on packing your specimen for shipment after collection and mixing.
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