Identifying subpopulations in multicellular systems by quantitative chemical imaging using label-free hyperspectral...

Area of Science:

• Biomedical Optics
• Chemical Imaging
• Microscopy

Background:

• Quantitative hyperspectral coherent Raman scattering microscopy offers label-free, non-invasive chemical analysis.
• It provides spectrally and spatially resolved chemical information with sub-cellular resolution.
• This technique has the potential to advance the study of complex living multicellular systems.

Purpose of the Study:

• To apply quantitative hyperspectral CARS microscopy and data analysis to image living mouse liver organoids and xenografted mouse brain tissue.
• To demonstrate the method's capability in discriminating cellular sub-populations based on chemical content.
• To validate the findings using correlative fluorescence microscopy.

Main Methods:

• Utilized an in-house developed hyperspectral CARS microscope.
• Employed a quantitative data analysis pipeline for unsupervised analysis of chemical content.
• Performed correlative fluorescence microscopy for validation.

Main Results:

• Successfully discriminated different cellular sub-populations in liver organoids and brain tissue based on chemical composition.
• Identified cells in different cell cycle phases within organoids.
• Distinguished normal brain tissue from glioblastoma, and cancer stem cell-derived tumors from non-stem glioblastoma cells.

Conclusions:

• Quantitative hyperspectral CARS microscopy is effective for label-free chemical imaging of multicellular systems.
• The method enables identification of distinct cellular sub-populations in biologically relevant samples.
• This approach offers new opportunities for non-invasive disease diagnostics.