Unified Vibrational and Multiphoton Label-Free Nonlinear Microscopy for Simultaneous Chemical and Structural Imaging.

Area of Science:

• Biomedical Optics
• Label-free Imaging
• Microscopy

Background:

• Nonlinear microscopy offers label-free imaging by utilizing intrinsic spectroscopic responses.
• Current clinical systems often use either multiphoton or vibrational contrast, limiting comprehensive analysis.
• This limits the diagnostic utility of label-free nonlinear microscopes for heterogeneous specimens.

Purpose of the Study:

• To develop a nonlinear microscopy strategy combining both multiphoton and vibrational contrasts.
• To create a robust, complementary contrast palette for enhanced morphofunctional analysis.
• To engineer a versatile biophotonics tool for potential clinical applications.

Main Methods:

• Systematic investigation of readout schemes to tailor the detection chain for quantitative performance.
• Development of a compact laser source exciting both vibrational coherences and multiphoton signals.
• Validation using rodent models of naturally occurring and allogeneic mammary cancer.

Main Results:

• A novel imaging platform successfully integrates multiphoton and vibrational contrast modalities.
• Engineering criteria were established for optimizing detection chains in nonlinear microscopy.
• The platform demonstrated effectiveness in imaging complex biological specimens, including cancers.

Conclusions:

• The proposed strategy enables a robust, complementary contrast palette in label-free nonlinear microscopy.
• The developed compact laser source and detection strategy enhance quantitative performance.
• This versatile and cost-effective biophotonics tool shows promise for widespread clinical adoption.