Expansion microscopy of mouse brain organotypic slice cultures to study protein distribution
- Highlights • Enables immunostaining and visualization of epitopes deep within brain slices • Utilizes expansion microscopy to increase imaging resolution • Optimized for brain organotypic slice cultures and tested in acute brain slices • Analysis workflow for protein distribution (surface vs. intracellular pool) using Imaris Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Summary Assessing protein distribution with super-resolution in tissue is often complicated and restrictive. Here, we describe a protocol for immunostaining and expansion microscopy imaging of mouse brain organotypic slice cultures. We detail an Imaris analysis workflow to analyze the surface vs intracellular distribution of AMPA receptors at super-resolution during homeostatic plasticity. We have optimized the protocol for brain organotypic slice culture and tested in acute brain slices. This protocol is suitable to study protein distribution under multiple plasticity paradigms.
Author: | Diane BissenORCiDGND, Maximilian Ken Kracht, Franziska FoßGND, Amparo Acker-PalmerORCiDGND |
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URN: | urn:nbn:de:hebis:30:3-783663 |
DOI: | https://doi.org/10.1016/j.xpro.2022.101507 |
ISSN: | 2666-1667 |
Parent Title (English): | STAR Protocols |
Publisher: | Elsevier |
Place of publication: | Amsterdam |
Document Type: | Article |
Language: | English |
Date of Publication (online): | 2022/06/24 |
Date of first Publication: | 2022/06/24 |
Publishing Institution: | Universitätsbibliothek Johann Christian Senckenberg |
Release Date: | 2024/05/16 |
Tag: | Cell Biology; Microscopy; Neuroscience |
Volume: | 3 |
Issue: | 3, 101507 |
Article Number: | 101507 |
Page Number: | 25 |
First Page: | 1 |
Last Page: | 24 |
HeBIS-PPN: | 520982495 |
Institutes: | Biowissenschaften / Biowissenschaften |
Fachübergreifende Einrichtungen / Buchmann Institut für Molekulare Lebenswissenschaften (BMLS) | |
Dewey Decimal Classification: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
Sammlungen: | Universitätspublikationen |
Licence (German): | ![]() |