Mammalian cell and tissue imaging using Raman and coherent Raman microscopy | JoVE Visualize

Area of Science:

Molecular and Cellular Biology
Biophysics
Analytical Chemistry

Background:

Direct imaging of cellular and tissue metabolism is crucial for understanding biological processes.
Raman scattering (RS) microscopy, especially coherent Raman scattering (CRS) techniques like CARS and SRS, provides label-free, chemically selective imaging.
RS microscopy is valuable for identifying subcellular structures, observing metabolism, and characterizing phenotypes.

Purpose of the Study:

To overview Raman scattering modalities and techniques for mammalian cell and tissue imaging.
To focus on the advances and applications of coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS) microscopy.
To enhance understanding of the metabolism and dynamics of key biomolecules (lipids, protein, glucose, nucleic acids) in mammalian systems.

Main Methods:

Review of Raman scattering (RS) microscopy techniques, emphasizing coherent Raman scattering (CRS) modalities.
Detailed examination of coherent anti-Stokes Raman scattering (CARS) and stimulated Raman scattering (SRS) microscopy.
Discussion of combining RS with other techniques (fluorescence, IR spectroscopy, flow cytometry, RNA-sequencing) to expand applications.

Main Results:

CARS and SRS microscopy demonstrate high chemical selectivity, sensitivity, and imaging speed for biological samples.
These techniques enable direct visualization and quantification of metabolic components like lipids, proteins, glucose, and nucleic acids.
Integration with other methods broadens the scope of RS imaging in diverse fields such as microbiology, systems biology, neurology, and tumor biology.

Conclusions:

Coherent Raman scattering microscopy, particularly CARS and SRS, represents a powerful tool for direct metabolic imaging in cells and tissues.
These advanced imaging techniques facilitate a deeper understanding of molecular dynamics and metabolic processes in mammalian systems.
The versatility of RS microscopy, especially when conjugated with other analytical methods, offers significant potential for future biological and medical research.