INSPIRE: Single-beam probed complementary vibrational bioimaging.

Area of Science:

• Biomedical Optics
• Spectroscopy
• Chemical Imaging

Background:

• Molecular spectroscopy, including stimulated Raman imaging, provides intrinsic contrast for in situ chemical analysis of biomolecules.
• Limitations exist due to Raman inactive or weak vibrational modes, hindering broader applications.
• Infrared (IR) spectroscopy offers complementary spectral information, but integrating it with Raman is optically challenging.

Purpose of the Study:

• To introduce a novel cross-modality spectroscopic imaging approach, INSPIRE (in situ pump-probe IR and Raman excitation) microscopy.
• To overcome the optical challenges of combining IR and Raman spectroscopy for simultaneous detection.
• To enable high-content molecular imaging with enhanced sensitivity and specificity.

Main Methods:

• Development of INSPIRE microscopy, encoding ultrafast Raman and IR photothermal relaxation into a single probe beam.
• Simultaneous detection of both Raman and IR signals.
• Application of the technique to imaging of chemicals, cells, tissues, and organisms.

Main Results:

• INSPIRE microscopy successfully integrates complementary spectral information from IR and Raman spectroscopy.
• Demonstrated high-content molecular imaging capabilities across various biological samples.
• Showcased applicability to label-free and molecular tag imaging.

Conclusions:

• INSPIRE microscopy is a powerful new tool for in situ chemical imaging, overcoming limitations of individual techniques.
• The approach offers significant potential for label-free optical sensing and imaging in biomedicine and materials science.
• This cross-modality technique enhances molecular information retrieval for complex biological systems.