Label-free photothermal optical coherence microscopy to locate desired regions of interest in multiphoton imaging of volumetric specimens

Biochip-based research is currently evolving into a three-dimensional and large-scale basis similar to the in vivo microenvironment. For the long-term live and high-resolution imaging in these specimens, nonlinear microscopy capable of label-free and multiscale imaging is becoming increasingly impor...

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Bibliographic Details
Published in:Scientific reports 2023-03, Vol.13 (1), p.3625-3625, Article 3625
Main Authors: Ravichandran, Naresh Kumar, Hur, Hwan, Kim, Hyemi, Hyun, Sangwon, Bae, Ji Yong, Kim, Dong Uk, Kim, I Jong, Nam, Ki-Hwan, Chang, Ki Soo, Lee, Kye-Sung
Format: Article
Language:English
Subjects:
639/624/1107/328/1650
639/624/1107/510
639/624/1111/55
Biological samples
Humanities and Social Sciences
Microenvironments
Microscopy
Movement
multidisciplinary
Phantoms, Imaging
Science
Science (multidisciplinary)
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Summary:Biochip-based research is currently evolving into a three-dimensional and large-scale basis similar to the in vivo microenvironment. For the long-term live and high-resolution imaging in these specimens, nonlinear microscopy capable of label-free and multiscale imaging is becoming increasingly important. Combination with non-destructive contrast imaging will be useful for effectively locating regions of interest (ROI) in large specimens and consequently minimizing photodamage. In this study, a label-free photothermal optical coherence microscopy (OCM) serves as a new approach to locate the desired ROI within biological samples which are under investigation by multiphoton microscopy (MPM). The weak photothermal perturbation in sample by the MPM laser with reduced power was detected at the endogenous photothermal particles within the ROI using the highly sensitive phase-differentiated photothermal (PD–PT) OCM. By monitoring the temporal change of the photothermal response signal of the PD–PT OCM, the hotspot generated within the sample focused by the MPM laser was located on the ROI. Combined with automated sample movement in the x–y axis, the focal plane of MPM could be effectively navigated to the desired portion of a volumetric sample for high-resolution targeted MPM imaging. We demonstrated the feasibility of the proposed method in second harmonic generation microscopy using two phantom samples and a biological sample, a fixed insect on microscope slide, with dimensions of 4 mm wide, 4 mm long, and 1 mm thick.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-023-30524-z