Simultaneous label-free autofluorescence multi-harmonic microscopy driven by the supercontinuum generated from a bulk...

Area of Science:

• Biomedical Optics
• Microscopy
• Laser Physics

Background:

• Nonlinear microscopy uses lasers to image intrinsic molecules in biological samples.
• Current systems have limited spectral capabilities, requiring multiple lasers for simultaneous molecular analysis.
• This increases cost and complexity for applications like digital histopathology and image-guided surgery.

Purpose of the Study:

• To develop a compact and practical supercontinuum laser source for nonlinear multiphoton microscopy.
• To enable simultaneous, label-free imaging of multiple endogenous molecules in biological tissues.
• To provide a cost-effective alternative for clinical translation of nonlinear microscopy.

Main Methods:

• Utilized supercontinuum generation from a bulk yttrium aluminum garnet (YAG) crystal.
• Generated broadband femtosecond radiation covering visible and near-infrared spectra.
• Applied the generated supercontinuum to nonlinear multiphoton microscopy for imaging.

Main Results:

• The supercontinuum source provided broadband radiation (550-900 nm and 950-1200 nm).
• Pulses within 1030-1150 nm enabled label-free volumetric chemical imaging of ex vivo chinchilla kidney.
• Demonstrated simultaneous autofluorescence multi-harmonic imaging capability.

Conclusions:

• Supercontinuum generation from bulk YAG crystals is a powerful source for multimodal nonlinear microscopy.
• The developed system offers a practical and compact solution for advanced biological imaging.
• This approach facilitates the clinical translation of label-free nonlinear microscopy techniques.