Cell optical density and molecular composition revealed by simultaneous multimodal label-free imaging

Area of Science:

• Biophotonics
• Cellular Imaging
• Spectroscopy

Background:

• Label-free cellular analysis is crucial for understanding cell dynamics.
• Raman imaging and digital holographic microscopy (DHM) offer complementary information.
• Integrating these techniques can enhance cellular characterization.

Purpose of the Study:

• To combine Raman imaging with simultaneous phase measurements for unlabeled cell analysis.
• To correlate chemical information from Raman spectroscopy with quantitative phase information from DHM.
• To demonstrate the utility of this multimodal approach for cell composition and dynamics studies.

Main Methods:

• Simultaneous acquisition of Raman imaging and digital holographic microscopy data.
• Analysis of light retardation and scattering by cellular molecules.
• Quantification of spatial correlation between Raman and phase images.
• Video-rate imaging for observing dynamic cellular changes.

Main Results:

• Demonstrated the correlation between cellular chemistry (Raman) and physical properties (phase).
• Showcased the complementary nature of label-free data from different scattering mechanisms.
• Observed dynamic cellular changes during Raman acquisition using video-rate imaging.
• Validated the multimodal approach for comprehensive cell analysis.

Conclusions:

• The combined Raman imaging and DHM approach provides complementary label-free information on cell composition and dynamics.
• This multimodal technique offers enhanced insights not achievable by individual methods.
• The method allows for real-time observation of cellular changes during measurement.