Intravital imaging by simultaneous label-free autofluorescence-multiharmonic microscopy

Sixian You, Haohua Tu (), Eric J. Chaney, Yi Sun, Youbo Zhao, Andrew J. Bower, Yuan-Zhi Liu, Marina Marjanovic, Saurabh Sinha, Yang Pu and Stephen A. Boppart ()
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Sixian You: University of Illinois at Urbana-Champaign
Haohua Tu: University of Illinois at Urbana-Champaign
Eric J. Chaney: University of Illinois at Urbana-Champaign
Yi Sun: University of Illinois at Urbana-Champaign
Youbo Zhao: University of Illinois at Urbana-Champaign
Andrew J. Bower: University of Illinois at Urbana-Champaign
Yuan-Zhi Liu: University of Illinois at Urbana-Champaign
Marina Marjanovic: University of Illinois at Urbana-Champaign
Saurabh Sinha: University of Illinois at Urbana-Champaign
Yang Pu: University of Illinois at Urbana-Champaign
Stephen A. Boppart: University of Illinois at Urbana-Champaign

Nature Communications, 2018, vol. 9, issue 1, 1-9

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-04470-8

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