用于荧光标记的离体大鼠脑片光学透明化和成像的方案。

Protocol for optical clearing and imaging of fluorescently labeled ex vivo rat brain slices.

机构信息

Developmental and Regenerative Neurobiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Department of Experimental Medical Science, Lund University, Lund, Sweden.

Developmental and Regenerative Neurobiology, Wallenberg Neuroscience Center, Lund Stem Cell Center, Department of Experimental Medical Science, Lund University, Lund, Sweden; Center for Neuroscience, University of Copenhagen Faculty of Health and Medical Sciences, Copenhagen, Denmark.

出版信息

STAR Protoc. 2023 Mar 17;4(1):102041. doi: 10.1016/j.xpro.2022.102041. Epub 2023 Jan 18.

DOI:10.1016/j.xpro.2022.102041

PMID:36853668

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9873499/

Abstract

Tissue clearing is commonly used for whole-brain imaging but seldom used for brain slices. Here, we present a simple protocol to slice, immunostain, and clear sections of adult rat brains for subsequent high-resolution confocal imaging. The protocol does not require toxic reagents or specialized equipment. We also provide instructions for culturing of rat brain slices free floating on permeable culture inserts, maintained in regular CO incubators, and handled only at media change.

摘要

组织透明化通常用于全脑成像，但很少用于脑切片。在这里，我们提出了一种简单的方案，用于对成年大鼠脑切片进行免疫染色和透明化，以便随后进行高分辨率共聚焦成像。该方案不需要有毒试剂或特殊设备。我们还提供了在透气培养插入物上自由漂浮培养大鼠脑切片的说明，这些切片可以在常规 CO 培养箱中维持，并仅在更换培养基时进行处理。