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1. Can the liquid volume in cryobags exceed the recommended capacity?

No. Exceeding the recommended fill volume may lead to:

  • Risk of rupture: Liquid expansion during freezing can cause cracks or leaks.
  • Sealing failure: Overfilling may compromise the closure, leading to contamination.
  • Storage inefficiency: Overfilled bags hinder stacking and occupy excess space.
    Always adhere to the labeled capacity for optimal performance.

Degassing ensures:

  • Reduced ice crystal formation, minimizing cell damage.
  • Secure sealing by eliminating air pockets.
  • Uniform freezing and prevention of bag rupture.
  • Higher cell viability post-thaw.
    Always expel air before sealing to maximize sample integrity.
  • Adjust temperature/time based on material (PVC, EVA).
  • Apply even pressure for consistent seals.
  • Clean and dry the sealing area to avoid residue interference.
  • Align edges precisely before sealing.
  • Use validated equipment (e.g., RF heat sealers).
  • Inspect seals post-process for leaks.
  • Use controlled-rate freezing:
    • Initial cooling rate: ​~1°C per minute (or per protocol).
    • Avoid abrupt temperature drops to prevent thermal shock.
  • Employ programmable freezers for precision.
  • Never place bags directly into liquid nitrogen without gradual cooling.
    Slow cooling minimizes ice crystal damage and maintains cell viability.
  • Rapid thawing: Submerge in ​37°C water bath (≤40°C) for ​2–3 minutes until ice melts.
  • Avoid water contact: Use a sealed secondary bag if necessary.
  • Gentle agitation: Swirl gently to ensure even warming.
  • Aseptic transfer: Move thawed samples to sterile conditions immediately.
  • Inspect integrity: Check for leaks before use.
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