Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic...

Area of Science:

• Biomedical Optics
• Microscopy
• Histology

Background:

• Standard histology relies on exogenous stains for tissue visualization.
• Multicolor nonlinear microscopy offers a stain-free alternative for imaging living tissues.
• Understanding intrinsic optical properties is key for advanced microscopy applications.

Purpose of the Study:

• To demonstrate multicolor nonlinear microscopy for high-resolution, stain-free imaging of living tissues.
• To highlight the use of intrinsic fluorescence and second harmonic generation for biological imaging.
• To showcase applications in visualizing tissue morphology, cell metabolism, and disease states.

Main Methods:

• Utilizing two- and three-photon-excited intrinsic fluorescence.
• Employing second harmonic generation from supermolecular structures.
• Imaging endogenous molecules like nicotinamide adenine dinucleotide, retinol, indoleamines, and collagen.

Main Results:

• Achieved imaging resolution and detail comparable to standard histology without exogenous stains.
• Successfully visualized tissue morphology, cell metabolism, and disease states.
• Demonstrated the capability to image a range of intrinsic molecules and molecular assemblies.

Conclusions:

• Multicolor nonlinear microscopy with intrinsic contrast is a powerful tool for biological imaging.
• This technique provides crucial physiological and pathological information without the need for stains.
• Applications span from basic research to disease diagnostics, including Alzheimer's disease and cancer.