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

摘要: Rapid cooling of aqueous solutions is a useful approach for two important biological applications: (I) cryopreservation cells and tissues long-term storage, (II) cryofixation ultrastructural investigations by electron cryo-electron microscopy. Usually, both approaches are very different in methodology. Here we show that novel, fast easy to use technique called self-pressurized rapid freezing (SPRF) is-after some adaptations-also versatile cryopreservation. Sealed metal tubes with high thermal diffusivity containing the samples plunged into liquid cryogen. Internal pressure builds up reducing ice crystal formation therefore supporting reversible through vitrification cells. After rewarming pressurized samples, viability rates > 90% can be reached, comparable best-performing established devices tested. In addition, small SPRF allow space-saving sample storage sealed containers prevent contamination from or cryogen during freezing, thawing.