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Non-Uniform Distribution of Cryoprotecting Agents in Rice Culture Cells Measured by CARS Microscopy.
Fionna M D Samuels1, Dominik G Stich2, Remi Bonnart3
1Department of Chemistry, Colorado State University, Fort Collins, CO 80523, USA.
Plants (Basel, Switzerland)
|April 3, 2021
Summary
Cryoprotectants like DMSO are crucial for cell cryopreservation. New imaging reveals deuterated DMSO (d6-DMSO) concentrates in specific cell organelles, not uniformly, impacting cell survival during freezing.
Area of Science:
- Cell biology
- Biophysics
- Microscopy
Background:
- Cryoprotective agents (CPAs) are essential for preserving cells via cryopreservation.
- The precise mechanisms by which CPAs ensure cell viability during freezing and thawing remain incompletely understood.
- Optimizing CPA formulations for diverse cell types is challenging due to limited knowledge of their intracellular behavior.
Purpose of the Study:
- To visualize the intracellular distribution of a deuterated cryoprotectant within living cells.
- To investigate the localization of deuterated dimethyl sulfoxide (d6-DMSO) in rice suspension cells.
- To gain insights into how cryoprotectant distribution affects cellular cryopreservation.
Main Methods:
- Coherent anti-Stokes Raman scattering (CARS) microscopy was employed for live-cell imaging.
- Deuterated dimethyl sulfoxide (d6-DMSO) was used as the model cryoprotectant.
- Intracellular localization of d6-DMSO within rice suspension cells was directly visualized.
Main Results:
- d6-DMSO exhibited non-uniform distribution within rice suspension cells.
- d6-DMSO was observed to sequester within specific cellular organelles.
- This compartmentalization alters the understanding of DMSO concentration gradients within cells.
Conclusions:
- Intracellular cryoprotectant distribution is heterogeneous and compartment-specific.
- Variations in CPA concentration within cellular compartments may influence cryoprotection efficacy.
- Further studies on diverse CPAs and mixtures are needed to establish comprehensive cryopreservation strategies.
