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Updated: Jan 31, 2026
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Direct Comparison of Hyperspectral Stimulated Raman Scattering and Coherent Anti-Stokes Raman Scattering Microscopy for Chemical Imaging
Published on: April 28, 2022
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Label-Free Volumetric Quantitative Imaging of the Human Somatic Cell Division by Hyperspectral Coherent Anti-Stokes
Arnica Karuna1, Francesco Masia1, Marie Wiltshire2
1School of Physics and Astronomy , Cardiff University , The Parade , Cardiff CF24 3AA , United Kingdom.
Analytical Chemistry
|January 10, 2019
Summary
Hyperspectral coherent anti-Stokes Raman scattering (CARS) microscopy enables quantitative 3D chemical imaging of dividing cells. This technique reveals cell composition and mass changes, aiding cell biology research.
Area of Science:
- Cell Biology
- Biophysics
- Chemical Imaging
Background:
- Quantitative chemical imaging of unstained cells is crucial for cell biology.
- Label-free optical microscopy offers non-invasive analysis but lacks chemical specificity.
- Advancing cell and tissue analytics requires high spatio-temporal resolution in 3D.
Purpose of the Study:
- To demonstrate hyperspectral coherent anti-Stokes Raman scattering (CARS) microscopy for quantitative 3D chemical imaging of human osteosarcoma cells.
- To develop and apply a quantitative data analysis method for volumetric chemical composition analysis.
- To investigate the impact of cell division and drug perturbation on cellular chemical composition and mass.
Main Methods:
- Hyperspectral coherent anti-Stokes Raman scattering (CARS) microscopy was employed for label-free imaging.
- A quantitative data analysis method was developed to generate 3D chemical composition maps (water, proteins, DNAP, lipids).
- Correlative two-photon fluorescence microscopy and specific fluorescent probes were used for validation.
Main Results:
- Quantitative volumetric 3D images of dividing human osteosarcoma cells were successfully generated.
- The dry masses of organic components (proteins, DNAP, lipids) were determined.
- Perturbation of cell division with Taxol showed significant changes in cellular organization and component masses.
Conclusions:
- Hyperspectral CARS microscopy provides a powerful tool for quantitative chemical imaging of cells in 3D.
- The method allows for the determination of cellular dry mass and the study of chemical changes during cell division and drug treatment.
- This technique advances cell analytics and provides insights into cell cycle dynamics and drug effects.
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