This prevents cells from being lost due to purification and that a lot of time has to be spent on laborious searches in the laboratory. At the same time, the sperm can be used for the ICSI immediately after thawing without major preparation.
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Learn more about the background and handling of SpermVD in this interesting webinar from Gynemed and MFC Global with Dr. Arie Berkovitz and Miki Belenky.
If you have any further questions, please do not hesitate to contact us.
In a Petri dish, individual sperm are first collected in small drops of sperm washing medium (eg GM501 SpermAir) under oil coating.
Using sterile forceps remove the SpermVD from the packaging under sterile conditions and place on a sterile surface (e.g. a Petri dish or Petri dish lid).
Prepare a working solution of sperm freezing medium (from washing and freezing medium, e.g. 30 μl GM501 SpermAir and 21 μl GM501 SpermStore (1: 0.7). for the freezing solution. Three 0.5-1 μl drops are added to the appropriate wells of SpermVD. Immediately transfer the SpermVD to the Petri dish (item 1). It should be noted that the drops must be covered with oil.
Using a micromanipulator and an ICSI pipette pre-filled with PVP solution, the individual sperm are transferred to one or more freezing drops.
Freeze the loaded SpermVD with the sperm within 10 min of adding the first cell.
Using a micromanipulator and an ICSI pipette pre-filled with PVP solution, the individual sperm are transferred to one or more freezing drops. Freeze the loaded SpermVD with the sperm within 10 min of adding the first cell. Using sterile tweezers, carefully dispense the SpermVD with the sperm from the Petri dish (allowing the excess oil to drain well) into a cryo-safe labeled 1.8-3.6 ml cryotube. Recommendation: do not close the tube tightly so that the liquid nitrogen can gradually penetrate and fill the tube promptly after being placed in the liquid nitrogen, thereby allowing safe handling of the cryotubes with the SpermVD. Transfer the cryotube with SpermVD directly into the liquid nitrogen for long-term storage.
Prepare a Petri dish (50-100 mm) with PVP drops and sperm wash medium (e.g. GM501 SpermAir) drops with oil overlay (transfer dish).
Bring the cryotube directly into room temperature with the SpermVD liquid nitrogen and leave the tube unopened for 5 min at room temperature.
Remove the SpermVD from the cryotube with sterile forceps and place in the Transfer Bowl. The drops with the sperm must be covered with oil.
Using a micromanipulator and an ICSI pipette, transfer the sperm into the sperm washing medium drops at 20x magnification from the SpermVD.
- 2020 Systems Biology in Reproductive medicine, M. Belenky, D. Itzhakov, V. Freger, O. Roseman, S. Abehsera, N. Miller, A. Berkovitz “Optimizing the protocol for vitrification of individual spermatozoa by adjusting equilibration”
- 2018 Nov Hum Reprod. 1;33(11):1975-1983 A. Berkovitz, N. Miller, M. Silberman, M. Belenky and P. Itsykson “A novel solution for freezing small numbers of spermatozoa using a sperm vitrification device”
- 2017 ESHRE (P-032) A. Berkovitz; M. Belenky and P. Itsykson “A novel solution for freezing small numbers of spermatozoa by a sperm vitrification device”