Expansion microscopy of mouse brain organotypic slice cultures to study protein distribution

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 intracellul...

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Published inSTAR protocols Vol. 3; no. 3; p. 101507
Main Authors Bissen, Diane, Kracht, Maximilian Ken, Foss, Franziska, Acker-Palmer, Amparo
Format Journal Article
LanguageEnglish
Published Elsevier Inc 16.09.2022
Elsevier
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Cell Biology
Microscopy
Neuroscience
Protocol
Microscopy
Neuroscience
Cell Biology
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Abstract 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. For complete details on the use and execution of this protocol, please refer to Bissen et al. (2021). [Display omitted] •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. 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.
AbstractList 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.For complete details on the use and execution of this protocol, please refer to Bissen et al. (2021). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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. For complete details on the use and execution of this protocol, please refer to Bissen et al. (2021) . • 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. 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.
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. For complete details on the use and execution of this protocol, please refer to Bissen et al. (2021). [Display omitted] •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. 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.
ArticleNumber 101507
Author Foss, Franziska
Kracht, Maximilian Ken
Bissen, Diane
Acker-Palmer, Amparo
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Snippet Assessing protein distribution with super-resolution in tissue is often complicated and restrictive. Here, we describe a protocol for immunostaining and...
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SubjectTerms Cell Biology
Microscopy
Neuroscience
Protocol
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Title Expansion microscopy of mouse brain organotypic slice cultures to study protein distribution
URI https://dx.doi.org/10.1016/j.xpro.2022.101507
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