Institute of Molecular Medicine, Ulm University, 89081 Ulm, Germany.
Stem Cell Aging Group, Regenerative Medicine Program, The Bellvitge Institute for Biomedical Research (IDIBELL), L'Hospitalet de Llobregat, Barcelona, Spain; Program for Advancing the Clinical Translation of Regenerative Medicine of Catalonia, P-CMR[C], L'Hospitalet de Llobregat, Barcelona, Spain.
STAR Protoc. 2022 Jun 17;3(3):101483. doi: 10.1016/j.xpro.2022.101483. eCollection 2022 Sep 16.
Quantitative 3D imaging of organ-wide cellular and subcellular components is central for revealing and understanding complex interactions between stem cells and their microenvironment. Here, we present a gentle but fast whole-mount immunofluorescence staining protocol for 3D confocal microscopy (iFAST3D) that preserves the 3D structure of the entire tissue and that of subcellular structures with high fidelity. The iFAST3D protocol enables reproducible and high-resolution 3D imaging of stem cells and various niche components for many mouse organs and tissues. For complete details on the use and execution of this protocol, please refer to Saçma et al. (2019).
对器官范围的细胞和亚细胞成分进行定量 3D 成像,对于揭示和理解干细胞与其微环境之间的复杂相互作用至关重要。在这里,我们提出了一种温和但快速的用于 3D 共聚焦显微镜的全组织免疫荧光染色方案(iFAST3D),该方案可以高保真度地保留整个组织和亚细胞结构的 3D 结构。iFAST3D 方案可实现许多小鼠器官和组织中干细胞和各种壁龛成分的可重复和高分辨率 3D 成像。有关此方案的使用和执行的完整详细信息,请参阅 Saçma 等人(2019 年)。