Reversible Cryopreservation of Living Cells Using an Electron Microscopy Cryo-Fixation Method

Area of Science:

• Biophysics
• Cryobiology
• Microscopy

Background:

• Rapid cooling is crucial for cryopreservation and cryofixation.
• Established methods for these applications often differ significantly in technique.

Purpose of the Study:

• To demonstrate the adaptability of self-pressurized rapid freezing (SPRF) for cryopreservation.
• To evaluate SPRF's efficacy in achieving high cell viability and vitrification.

Main Methods:

• Utilizing sealed metal tubes with high thermal diffusivity for sample containment.
• Plunging samples into liquid cryogen to induce self-pressurization and rapid cooling.
• Employing rapid rewarming techniques for sample recovery.

Main Results:

• SPRF, with adaptations, proved effective for cryopreservation.
• Achieved cell viability rates exceeding 90%, comparable to established methods.
• Vitrification of cells was supported by reduced ice crystal formation due to internal pressure.
• Sealed containers prevented cryogen contamination and allowed space-saving storage.

Conclusions:

• SPRF is a versatile and efficient technique for both cryofixation and cryopreservation.
• The method offers advantages in terms of viability, contamination prevention, and storage.
• SPRF presents a promising alternative for biological sample preservation.