Non-Uniform Distribution of Cryoprotecting Agents in Rice Culture Cells Measured by CARS Microscopy.

Citation
Samuels, F. M. D., et al. “Non-Uniform Distribution Of Cryoprotecting Agents In Rice Culture Cells Measured By Cars Microscopy.”. Plants (Basel, Switzerland).
Center University of Colorado Denver
Author Fionna M D Samuels, Dominik G Stich, Remi Bonnart, Gayle M Volk, Nancy E Levinger
Keywords cryopreservation, cryoprotectant distribution, raman microscopy
Abstract

Cryoprotectants allow cells to be frozen in liquid nitrogen and cryopreserved for years by minimizing the damage that occurs in cooling and warming processes. Unfortunately, how the specific cryoprotectants keep the cells viable through the cryopreservation process is not entirely evident. This contributes to the arduous process of optimizing cryoprotectant formulations for each new cell line or species that is conserved. Coherent anti-Stokes Raman scattering microscopy facilitates the visualization of deuterated cryoprotectants within living cells. Using this technique, we directly imaged the location of fully deuterated dimethyl sulfoxide (d-DMSO), the deuterated form of a commonly used cryoprotectant, DMSO, within rice suspension cells. This work showed that d-DMSO does not uniformly distribute throughout the cells, rather it enters the cell and sequesters within organelles, changing our understanding of how DMSO concentration varies within the cellular compartments. Variations in cryoprotectant concentration within different cells and tissues will likely lead to differing protection from liquid nitrogen exposure. Expanding this work to include different cryoprotectants and mixtures of cryoprotectants is vital to create a robust understanding of how the distributions of these molecules change when different cryoprotectants are used.

Year of Publication
2021
Journal
Plants (Basel, Switzerland)
Volume
10
Issue
3
Date Published
03/2021
ISSN Number
2223-7747
DOI
10.3390/plants10030589
Alternate Journal
Plants (Basel)
PMID
33801034
PMCID
PMC8003852