2 Materials
1. Nitrile gloves.
2. Liquid nitrogen (LN 2 ) and LN 2 storage tank with sample
storage system.
3. Benchtop dewar flasks (2 L capacity).
4. Styrofoam box for sperm freezing (L: 25.4 cm, W: 22.7 cm, H:
15.2 cm inner dimensions).
5. Metal tube rack (5 cm in height).
6. Cryoprotective gear: cryo-apron, cryo-gloves, and unvented
safety goggles.
7. Cryo-label printer, cryo-canes, cryo-cane sleeves, and cryocane goblets.
8. 0.25 cc French straws (IMV 005565 or Minitube 13407/
0010).
9. 1 cc Monoject syringe.
10. Versi-Dry Lab Soaker for benchtop.
11. 70% ethanol in spray bottle.
12. Petri dishes (Falcon 35 Â 10 mm, Becton Dickinson, Catalog #
351008).
13. Disposal bags.
14. Kimwipes.
15. Slide warmer adjusted to 37
C.
16. Fine microscissors and forceps.
17. Curved, serrated forceps, hemostats, and tong.
18. 0.5 mL and 1.5 mL microcentrifuge tubes.
19. 20, 200, and 1000 μL capacity pipetman and pipette tips.
20. Microcentrifuge.
21. High-speed centrifuge with 4
C cooling capacity.
22. Hamilton-Thorne IVOS sperm analysis system and 80-μm
deep dual-sided chamber (2Â CELL; Hamilton Thorne
Biosciences).
23. Heat sealer.
24. Water bath.
25. Timer.
26. Osmometer.
27. Digital balance.
28. pH meter.
Mouse Sperm Cryopreservation
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1. Nitrile gloves.
2. Liquid nitrogen (LN 2 ) and LN 2 storage tank with sample
storage system.
3. Benchtop dewar flasks (2 L capacity).
4. Styrofoam box for sperm freezing (L: 25.4 cm, W: 22.7 cm, H:
15.2 cm inner dimensions).
5. Metal tube rack (5 cm in height).
6. Cryoprotective gear: cryo-apron, cryo-gloves, and unvented
safety goggles.
7. Cryo-label printer, cryo-canes, cryo-cane sleeves, and cryocane goblets.
8. 0.25 cc French straws (IMV 005565 or Minitube 13407/
0010).
9. 1 cc Monoject syringe.
10. Versi-Dry Lab Soaker for benchtop.
11. 70% ethanol in spray bottle.
12. Petri dishes (Falcon 35 Â 10 mm, Becton Dickinson, Catalog #
351008).
13. Disposal bags.
14. Kimwipes.
15. Slide warmer adjusted to 37
C.
16. Fine microscissors and forceps.
17. Curved, serrated forceps, hemostats, and tong.
18. 0.5 mL and 1.5 mL microcentrifuge tubes.
19. 20, 200, and 1000 μL capacity pipetman and pipette tips.
20. Microcentrifuge.
21. High-speed centrifuge with 4
C cooling capacity.
22. Hamilton-Thorne IVOS sperm analysis system and 80-μm
deep dual-sided chamber (2Â CELL; Hamilton Thorne
Biosciences).
23. Heat sealer.
24. Water bath.
25. Timer.
26. Osmometer.
27. Digital balance.
28. pH meter.
Mouse Sperm Cryopreservation
403