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Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells

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Published on: May 28, 2021

Isochoric supercooling cryomicroscopy.

Yuanheng Zhao¹, Leo Lou², Chenang Lyu³

¹Chinese Academy of Sciences Key Laboratory of Cryogenics, Technical Institute of Physics and Chemistry, Beijing, 100190, China; Department of Parasitology, Leiden University Center for Infectious Diseases (LU-CID), Leiden University Medical Center, Leiden, ZA, 2333, Netherlands; Department of Mechanical Engineering, University of California Berkeley, Berkeley, CA, 94720, USA.

|February 21, 2022

Summary

This summary is machine-generated.

A new isochoric supercooling cryomicroscope (ISCM) allows ice-free study of biological systems at subzero temperatures. This novel cryomicroscopy technique significantly improves cell survival rates during preservation.

Keywords:

Cryomicroscopy Cryopreservation HeLa cells Isochoric Supercooling

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Area of Science:

Biophysics
Cryobiology
Biochemistry

Background:

Studying biological systems at subzero temperatures is challenging due to ice formation.
Isochoric (constant-volume) conditions influence water's thermodynamic behavior below freezing.
Existing cryopreservation methods face limitations in preserving cellular integrity.

Purpose of the Study:

To introduce and validate an isochoric supercooling cryomicroscope (ISCM) for ice-free subzero studies.
To assess the stability of supercooled solutions within the ISCM.
To evaluate the efficacy of ISCM for biological sample preservation.

Main Methods:

Design and implementation of the isochoric supercooling cryomicroscope (ISCM).
Thermodynamic analysis of water behavior in isochoric systems at subfreezing temperatures.
Comparative study of HeLa cell survival in University of Wisconsin (UW) solution under conventional (+4°C) and isochoric supercooled (-5°C) conditions using fluorescence microscopy.

Main Results:

The ISCM successfully enables ice-free observation of biological systems at subzero temperatures and atmospheric pressure.
Supercooled solutions demonstrated stability within the ISCM.
HeLa cells stored at -5°C under isochoric supercooling showed similar survival to controls after three days, while +4°C storage resulted in approximately three times higher mortality.

Conclusions:

The ISCM is a viable technology for ice-free cryomicroscopy and biochemical studies at subzero temperatures.
Isochoric supercooling offers superior cell preservation compared to conventional storage at +4°C.
This technology holds promise for advancing biological matter preservation research.