Mammalian cell and tissue imaging using Raman and coherent Raman microscopy

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.