Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy

Nature Communications
Sixian YouStephen A Boppart

Abstract

Intravital microscopy (IVM) emerged and matured as a powerful tool for elucidating pathways in biological processes. Although label-free multiphoton IVM is attractive for its non-perturbative nature, its wide application has been hindered, mostly due to the limited contrast of each imaging modality and the challenge to integrate them. Here we introduce simultaneous label-free autofluorescence-multiharmonic (SLAM) microscopy, a single-excitation source nonlinear imaging platform that uses a custom-designed excitation window at 1110 nm and shaped ultrafast pulses at 10 MHz to enable fast (2-orders-of-magnitude improvement), simultaneous, and efficient acquisition of autofluorescence (FAD and NADH) and second/third harmonic generation from a wide array of cellular and extracellular components (e.g., tumor cells, immune cells, vesicles, and vessels) in living tissue using only 14 mW for extended time-lapse investigations. Our work demonstrates the versatility and efficiency of SLAM microscopy for tracking cellular events in vivo, and is a major enabling advance in label-free IVM.

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Citations

Oct 27, 2018·Journal of Cellular Physiology·Gary E CarverGary S Stein
Nov 17, 2019·Proceedings of the National Academy of Sciences of the United States of America·Sixian YouStephen A Boppart
Feb 10, 2019·Journal of Biomedical Optics·Weisi XieJonathan T C Liu
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Nov 22, 2018·Biomedical Optics Express·Sixian YouHaohua Tu
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Methods Mentioned

BETA
imaging technique

Software Mentioned

MATLAB script
FIJI Plugins
MATLAB

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