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Comprehensive single-shot biophysical cytometry using simultaneous quantitative phase imaging and Brillouin
Zachary A Steelman1, Zachary N Coker2, Anna Sedelnikova2
1Air Force Research Laboratory, JBSA Fort Sam Houston, San Antonio, TX, 78234, USA. zachary.steelman.1@us.af.mil.
Scientific Reports
|November 1, 2022
Summary
This study introduces a novel label-free image cytometry platform combining quantitative phase imaging and Brillouin spectroscopy. It simultaneously measures numerous biophysical properties of single cells, advancing cell biology research.
Area of Science:
- Biophysics
- Cell Biology
- Medical Diagnostics
Background:
- Traditional cytometry relies on fluorescent probes, limiting the measurement of crucial biophysical parameters.
- Cell density, refractive index, and viscosity are difficult to obtain with current methods.
Purpose of the Study:
- To develop a label-free image cytometry platform for simultaneous measurement of multiple biophysical properties.
- To overcome limitations of fluorescent probes in single-cell analysis.
Main Methods:
- Integration of quantitative phase imaging and Brillouin spectroscopy.
- Development of a geometric simplification for freshly plated cells.
- Label-free acquisition of over a dozen biophysical parameters.
Main Results:
- Simultaneous measurement of cellular diameter, volume, refractive index, mass density, non-aqueous mass, fluid volume, dry volume, fractional water content (by mass and volume), Brillouin shift, Brillouin linewidth, longitudinal modulus, longitudinal viscosity, loss modulus, and loss tangent.
- Validation across CHO-K1 cells, nocodazole-treated cells, and cells under hypoosmotic shock.
Conclusions:
- The developed platform offers a comprehensive, label-free approach to single-cell biophysical characterization.
- This technology opens new research avenues in biophysics, cell biology, and medicine.
