These Applications Notes are offered for guidance only. They have been supplied to us by DB1 users. However, we can offer no guarantees of their effectiveness. Biotronics Ltd accepts no liabilities for losses incurred by use of these methodologies.

Ovarian tissue freezing

Specimen

Slices or biopsies of ovarian cortical tissue.

Freeze Solution

OV FRZ#1 – 1.5 M DMSO

2ml DMSO + 18ml cryo buffer (SIVF cryo buffer from Cook IVF)

Procedure

    1. If large pieces of tissue have been received, use sterile fine nose scissors or a scalpel blade to cut tissue into 6mm x 1mm pieces.
    2. Transfer one or more pieces of tissue into each straw. Hold on ice for 10 min to ensure cryoprotectant permeation.
    3. At the end of the freeze program, plunge the straws into liquid nitrogen.

Programme

  1. 4 oC to -7oC at -3o/min
  2. Hold 5 min
  3. Seed
  4. Hold 5 min
  5. -7o to -40oC at -0.3o/min
  6. Freefall to -60oC
  7. END
  8. Plunge into liquid nitrogen
DB1 Embryofreeze
Expel and SARS