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Optical‐Resolution Photoacoustic Microscopy for Volumetric and Spectral Analysis of Histological and Immunochemical Samples
Author(s) -
Zhang Yu Shrike,
Yao Junjie,
Zhang Chi,
Li Lei,
Wang Lihong V.,
Xia Younan
Publication year - 2014
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201403812
Subject(s) - microscopy , resolution (logic) , chromogenic , materials science , sample preparation , photoacoustic imaging in biomedicine , optics , photoacoustic spectroscopy , immunostaining , biomedical engineering , chemistry , pathology , immunohistochemistry , computer science , chromatography , physics , artificial intelligence , medicine
Abstract Optical‐resolution photoacoustic microscopy (OR‐PAM) is an imaging modality with superb penetration depth and excellent absorption contrast. Here we demonstrate, for the first time, that this technique can advance quantitative analysis of conventional chromogenic histochemistry. Because OR‐PAM can quantify the absorption contrast at different wavelengths, it is feasible to spectrally resolve the specific biomolecules involved in a staining color. Furthermore, the tomographic capability of OR‐PAM allows for noninvasive volumetric imaging of a thick sample without microtoming it. By immunostaining the sample with different chromogenic agents, we further demonstrated the ability of OR‐PAM to resolve different types of cells in a coculture sample with imaging depths up to 1 mm. Taken together, the integration of OR‐PAM with (immuno)histochemistry offers a simple and versatile technique with broad applications in cell biology, pathology, tissue engineering, and related biomedical studies.