Rapid immunostaining method for three-dimensional volume imaging of biological tissues by magnetic force-induced focusin
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Rapid immunostaining method for three‑dimensional volume imaging of biological tissues by magnetic force‑induced focusing of the electric field Myeongsu Na1 · Kitae Kim1 · Hye Ryeong Lim3 · Chang Man Ha3 · Sunghoe Chang1,2 Received: 20 March 2020 / Accepted: 15 October 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Recent surges in tissue clearing technology have greatly advanced 3-dimensional (3D) volume imaging. Cleared tissues need to be stained with fluorescence probes for imaging but the current staining methods are too laborious and inefficient for thick 3D samples, which impedes the broad application of clearing technology. To overcome these limitations, we developed an advanced staining platform named EFIC in which a magnetic force focuses the electric field by bending it onto the sample. Such that EFIC applies a significantly lower electric field to maintain nanoscale structural integrity while effectively drives staining probes into pre-cleared 3D samples. We found that EFIC achieved a rapid and uniform staining of various proteins and vascular networks of the brain as well as other organs over the entire depth of imaging. EFIC stained tau deposits and the vascular structure in the post-mortem human brain of Alzheimer’s disease and intracerebral hemorrhage, respectively, enabling quantitative analysis. The effectiveness of EFIC was further extended by the successful staining of various targets in non-cleared 3D brain samples. Together, EFIC represents a versatile and reliable staining platform for rapidly analyzing 3D molecular signatures and can be applied to sectioning-free 3D histopathology for diagnostic purposes. Keywords Optical clearing · Tissue staining · 3D volume imaging · Focused electric field · Histopathological analysis
Introduction Recent advances in tissue clearing methods have improved our understanding of the organization of intact tissues (Chung and Deisseroth 2013; Jing et al. 2018; Susaki et al. 2015; Hama et al. 2015; Perbellini et al. 2017; Costantini et al. 2015). Most methods now achieve a comparable degree Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00429-020-02160-0) contains supplementary material, which is available to authorized users. * Sunghoe Chang [email protected] 1
Department of Physiology and Biomedical Sciences, Seoul National University College of Medicine, 103 Daehak‑ro, Jongno‑gu, Seoul 03080, South Korea
2
Neuroscience Research Institute, Seoul National University College of Medicine, 103 Daehak‑ro, Jongno‑gu, Seoul 03080, South Korea
3
Research Division and Brain Research Core Facility, Korea Brain Research Institute, Daegu 41068, South Korea
of tissue transparency, thus tissue opacity is no longer an obstacle to three-dimensional imaging of thick samples (Richardson and Lichtman 2015; Silvestri et al. 2016; Susaki and Ueda 2016). If the tissue does not express fluorescent proteins before clearing, it should be stained with fluorescent probes to be
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