骨骼肌转录本和蛋白质水平的空间分析。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2024-12-20 Epub Date: 2024-10-09 DOI:10.1016/j.xpro.2024.103378

Paola Pisterzi, Clara Martinez Mir, Ouafa Dahri, Isabel de Poorter, Sandra Batlles Parera, Milica Dostanić, Massimo Mastrangeli, Christine Mummery, Niels Geijsen, Fanny Sage

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引用次数: 0

摘要

骨骼肌空间分析揭示了意想不到的区域化基因表达模式，挑战了人们对肌肉是均质组织的认识。在此，我们介绍了一种对小鼠骨骼肌转录本和蛋白质水平进行空间分析的方案。我们介绍了胫骨前肌解剖、甲醛固定、组织切片机切割、杂交链反应（HCR）检测和扩增的步骤。然后我们详细介绍了免疫染色、组织清除和成像的步骤。该方案很容易适用于其他组织。

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Spatial analysis of transcript and protein levels in skeletal muscle.

Skeletal muscle spatial analyses have revealed unexpected regionalized gene expression patterns challenging the understanding of muscle as a homogeneous tissue. Here, we present a protocol for the spatial analysis of transcript and protein levels in murine skeletal muscle. We describe steps for tibialis anterior dissection, formaldehyde fixation, tissue chopper cutting, and hybridization chain reaction (HCR) detection and amplification. We then detail procedures for immunostaining, tissue clearing, and imaging. This protocol is easily adaptable to other tissues.

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