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Label-free Super-resolution Imaging Enabled by Vibrational Imaging of Swelled Tissue and Analysis

Miao, Kun and Lin, Li-En and Qian, Chenxi and Wei, Lu (2022) Label-free Super-resolution Imaging Enabled by Vibrational Imaging of Swelled Tissue and Analysis. Journal of Visualized Experiments (183). Art. No. e63824. ISSN 1940-087X. doi:10.3791/63824. https://resolver.caltech.edu/CaltechAUTHORS:20220913-219244900

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Abstract

The universal utilization of fluorescence microscopy, especially super-resolution microscopy, has greatly advanced knowledge about modern biology. Conversely, the requirement of fluorophore labeling in fluorescent techniques poses significant challenges, such as photobleaching and non-uniform labeling of fluorescent probes and prolonged sample processing. In this protocol, the detailed working procedures of vibrational imaging of swelled tissue and analysis (VISTA) are presented. VISTA circumvents obstacles associated with fluorophores and achieves label-free super-resolution volumetric imaging in biological samples with spatial resolution down to 78 nm. The procedure is established by embedding cells and tissues in hydrogel, isotropically expanding the hydrogel sample hybrid, and visualizing endogenous protein distributions by vibrational imaging with stimulated Raman scattering microscopy. The method is demonstrated on both cells and mouse brain tissues. Highly correlative VISTA and immunofluorescence images were observed, validating the protein origin of imaging specificities. Exploiting such correlation, a machine learning-based image-segmentation algorithm was trained to achieve multi-component prediction of nuclei, blood vessels, neuronal cells, and dendrites from label-free mouse brain images. The procedure was further adapted to investigate pathological poly-glutamine (polyQ) aggregates in cells and amyloid-beta (Aβ) plaques in brain tissues with high throughput, justifying its potential for large-scale clinical samples.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.3791/63824DOIArticle
ORCID:
AuthorORCID
Miao, Kun0000-0001-6567-3650
Lin, Li-En0000-0003-3086-6991
Qian, Chenxi0000-0003-4815-5565
Wei, Lu0000-0001-9170-2283
Issue or Number:183
DOI:10.3791/63824
Record Number:CaltechAUTHORS:20220913-219244900
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20220913-219244900
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:116907
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:04 Oct 2022 14:27
Last Modified:04 Oct 2022 14:27

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