Applying the Expansion Microscopy
Method in Neurobiology

Деревцова, К. З.; Pchitskaya, Ekaterina; Rakovskaya, A. V.; Bezprozvanny, Ilya

doi:10.1134/s0022093021030157

Cited by 3 publications

(1 citation statement)

References 26 publications

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“…Since its invention, ExM has had many useful applications in different research fields. The application of ExM in neuroscience was demonstrated to visualize synapses (Chen et al., 2015) in the first ExM studies, and ExM was further successfully applied to study neurons (Derevtsova et al., 2021; Gallagher & Zhao, 2021; Karagiannis & Boyden, 2018). One of the first demonstrations of protein ExM was in visualization of the synaptic pair of Bassoon and Homer1 (Chozinski et al., 2016).…”

Section: Commentarymentioning

confidence: 99%

Expansion Microscopy Application for Calcium Protein Clustering Imaging in Cells and Brain Tissues

et al. 2023

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Many biological studies require high-resolution imaging and subsequent analysis of cell organelles and molecules. Some membrane proteins form tight clusters, and this process is directly linked to their function. In most studies, these small protein clusters have been investigated by total internal reflection fluorescence (TIRF) microscopy, which enables imaging with high spatial resolution within 100 nm of the membrane surface. Recently developed expansion microscopy (ExM) makes it possible to achieve nanometer resolution using a conventional fluorescence microscope by physically expanding the sample. In this article, we describe implementation of ExM for imaging of protein clusters formed by the endoplasmic reticulum (ER) calcium sensor protein STIM1. This protein translocates during ER store depletion and forms clusters that support contact with plasma membrane (PM) calcium-channel proteins. ER calcium channels such as the type 1 inositol triphosphate receptor (IP3R) also form clusters, but their investigation by TIRF microscopy is impossible due to the large distance from the PM. In this article, we demonstrate how to investigate IP3R clustering using ExM in hippocampal brain tissues. We compare IP3R clustering in the CA1 area of the hippocampus of wild-type and 5xFAD Alzheimer's disease model mice. To facilitate future applications, we describe experimental protocols and image processing guidelines for application of ExM to membrane and ER protein clustering studies in cultured cells and brain tissues.

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Section: Commentarymentioning

confidence: 99%

Expansion Microscopy Application for Calcium Protein Clustering Imaging in Cells and Brain Tissues

et al. 2023

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Derivative Technologies of Expansion Microscopy and Applications in Biomedicine

Jia,

Cui,

Divsalar

et al. 2024

ChemBioChem

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Expansion microscopy (ExM) is an innovative super‐resolution imaging technique that utilizes physical expansion to magnify biological samples, facilitating the visualization of cellular structures that are challenging to observe using traditional optical microscopes. The fundamental principle of ExM revolves around employing a specialized hydrogel to uniformly expand biological samples, thereby achieving super‐resolution imaging under conventional optical imaging conditions. This technology finds application not only in various biological samples such as cells and tissue sections, but also enables super‐resolution imaging of large biological molecules including proteins, nucleic acids, and metabolite molecules. In recent years, numerous researchers have delved into ExM, resulting in the continuous development of a range of derivative technologies that optimize experimental protocols and broaden practical application fields. This article presents a comprehensive review of these derivative technologies, highlighting the utilization of ExM for anchoring nucleic acids, proteins, and other biological molecules, as well as its applications in biomedicine. Furthermore, this review offers insights into the future development prospects of ExM technology.

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