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Raman Cryomicroscopic Imaging and Sample Holder for Spectroscopic Subzero Temperature Measurements
Guanglin Yu1, Rui Li2, Allison Hubel3
1Department of Mechanical Engineering, University of Minnesota, Minneapolis, MN, USA.
Methods in Molecular Biology (Clifton, N.J.)
|August 16, 2020
Summary
Raman cryomicroscopy enables noninvasive analysis of single cells at subzero temperatures. This method visualizes intracellular ice formation and cryoprotectant distribution, crucial for cryopreservation research.
Area of Science:
- Biophysics
- Cell Biology
- Spectroscopy
Background:
- Raman spectroscopy offers noninvasive analysis of cellular properties.
- Low-temperature applications include ice detection and cryoprotectant distribution.
- Understanding cellular responses to freezing is vital for cryopreservation.
Purpose of the Study:
- To detail Raman cryomicroscopic imaging of single cells.
- To describe sample handling for subzero spectroscopic measurements.
- To present methods for analyzing intracellular ice formation and cryoprotectant partitioning.
Main Methods:
- Utilized a custom-built temperature-controlled cooling stage for Raman microscopy.
- Performed cryomicroscopic imaging on Jurkat cells cryopreserved in sucrose solution.
- Developed strategies for detecting intracellular ice formation (IIF).
Main Results:
- Demonstrated successful Raman cryomicroscopic imaging of Jurkat cells.
- Quantified sucrose distribution across the cell membrane.
- Identified and analyzed intracellular ice formation.
Conclusions:
- Raman cryomicroscopy is a powerful tool for studying cellular cryopreservation.
- The method allows for detailed analysis of cryoprotectant behavior and ice formation.
- This technique advances understanding of cell survival during freezing.
