2024-12-19["PIER_181_24111001.png","PIER_182_25012003.png","PIER_183_25052305.png","other\/special_issue_13.png"]
Confocal Microscopy with Optimized Excitation and Emission Wavelength for Ultradeep and Multi-Channel Bioimaging
By Tianxiang Wu
Weihang Geng
Yuhuang Zhang
Qiming Xia
Mingxi Zhang
Jin Li
Menglu Chen
Wang Xi
Shiyi Peng
Zhe Feng
Jun Qian
Progress In Electromagnetics Research, Vol. 180, 115-126, 2024
Abstract
The second near-infrared region (NIR-II, 900-1880 nm) spectral window has garnered significant attention in bioimaging due to its moderate light absorption, diminished photon scattering and reduced autofluorescence. Exploiting NIR-II fluorescence, confocal microscopy has achieved deep in vivo imaging. In this study, we have identified that the fluorescence with wavelength beyond 1400 nm offers superior imaging quality for NIR-II confocal microscopy, irrespective of the laser excitation source being continuous-wave or pulsed. Furthermore, leveraging the multiphoton excitation capabilities of femtosecond laser, we have successfully integrated multiphoton excited visible fluorescence channels into the NIR-II fluorescence confocal microscopic system. We have successfully employed this novel system to acquire up to six distinct fluorescence microscopic imaging channels with negligible cross-channel interference, as well as multi-channel and large-depth in vivo observation of mouse brain and kidney.