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ECM 凝胶包埋神经支配的胰腺类器官整体免疫荧光染色方案。

Protocol for whole-mount immunofluorescence staining of ECM gel-embedded innervated pancreatic organoids.

机构信息

Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany; Department of Histology and Embryology, Faculty of Medicine, Hitit University, Çorum, Turkey.

Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.

出版信息

STAR Protoc. 2024 Jun 21;5(2):103132. doi: 10.1016/j.xpro.2024.103132. Epub 2024 Jun 13.

DOI:10.1016/j.xpro.2024.103132
PMID:38875112
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11227007/
Abstract

The mandatory usage of extracellular matrix (ECM) gels in 3D cultures limits antibody penetration and increases background, while the removal of ECM gel causes disruption of morphology and sample loss. These factors pose challenges to effective immune labeling-based staining. Here, we present a protocol for whole-mount immunofluorescence staining of gel-embedded pancreatic organoids. We describe steps for sample fixation, blocking, and antibody incubation. We detail procedures for washing antibodies and mounting.

摘要

在 3D 培养中强制使用细胞外基质 (ECM) 凝胶会限制抗体渗透并增加背景，而去除 ECM 凝胶会导致形态破坏和样品损失。这些因素给基于有效免疫标记的染色带来了挑战。在这里，我们提出了一种用于凝胶包埋的胰腺类器官的全组织免疫荧光染色的方案。我们描述了样品固定、封闭和抗体孵育的步骤。我们详细说明了抗体洗涤和装片的程序。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cb5/11227007/a34dc08a3060/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cb5/11227007/29eb77f73e97/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cb5/11227007/a34dc08a3060/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cb5/11227007/29eb77f73e97/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cb5/11227007/a34dc08a3060/gr3.jpg

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