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Interface-Sensitive Epi-Coherent Anti-Stokes Raman Scattering Microscopy for Imaging Cell Adhesion Dynamics
Mingmin Zhou1, Bin Dong1,2, Laura Lukov1
1James Tarpo Jr. and Margaret Tarpo Department of Chemistry, Purdue University, 560 Oval Dr., West Lafayette, Indiana 47907, United States.
Analytical Chemistry
|March 5, 2026
Summary
Coherent anti-Stokes Raman scattering (CARS) microscopy offers label-free imaging of cell adhesion dynamics. This technique precisely visualizes cell-substrate interactions and lipid droplets, revealing new insights into cell behavior during mitosis and spreading.
Area of Science:
- Cell Biology
- Biophysics
- Microscopy
Background:
- Understanding cell adhesion dynamics is crucial for cell migration and extracellular matrix interactions.
- Conventional microscopy methods lack chemical information or require fluorescence labeling for interfacial studies.
Purpose of the Study:
- To develop a label-free microscopy technique for studying cell adhesion dynamics at interfaces.
- To investigate the precision of epi-CARS in measuring cell-substrate distances and imaging lipid droplets.
Main Methods:
- Utilized homodyne epi-coherent anti-Stokes Raman scattering (epi-CARS) microscopy focused at interfaces.
- Employed a confocal epi-CARS system with a pinhole to reject out-of-focus reflections.
- Applied the method to study cell-substrate adhesion dynamics in mitotic cells.
Main Results:
- Epi-CARS demonstrated intrinsic interfacial selectivity with negative contrast at cell attachment areas, precisely measuring cell-substrate distances.
- The technique provided high contrast imaging of cellular lipid droplets.
- Observed adhesion site splitting during mitosis and outward development during postmitotic spreading.
- Distinguished homogeneous adhesion at leading edges and heterogeneous patterns at retracting edges after mitosis.
Conclusions:
- Epi-CARS microscopy is a powerful label-free tool for studying cell adhesion dynamics and lipid droplet distribution.
- The method offers high precision in measuring nanometer-scale distances at cell-substrate interfaces.
- Revealed dynamic changes in cell adhesion patterns during mitosis and cell spreading.
