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Fast and high-fidelity in situ 3D imaging protocol for stem cells and niche components for mouse organs and tissues.
Saçma, Mehmet; Matteini, Francesca; Mulaw, Medhanie A; Hageb, Ali; Bogeska, Ruzhica; Sakk, Vadim; Vollmer, Angelika; Marka, Gina; Soller, Karin; Milsom, Michael D; Florian, Maria Carolina; Geiger, Hartmut.
Affiliation
  • Saçma M; Institute of Molecular Medicine, Ulm University, 89081 Ulm, Germany. Electronic address: mehmet.sacma@uni-ulm.de.
  • Matteini F; 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.
  • Mulaw MA; Molecular Oncology Institute of Experimental Cancer Research, Medical Faculty, Ulm University, Ulm, Germany.
  • Hageb A; Institute of Molecular Medicine, Ulm University, 89081 Ulm, Germany.
  • Bogeska R; Division of Experimental Hematology, German Cancer Research Center (DKFZ) and Heidelberg Institute for Stem Cell Technology and Experimental Medicine (HI-STEM), Heidelberg, Germany.
  • Sakk V; Institute of Molecular Medicine, Ulm University, 89081 Ulm, Germany.
  • Vollmer A; Institute of Molecular Medicine, Ulm University, 89081 Ulm, Germany.
  • Marka G; Institute of Molecular Medicine, Ulm University, 89081 Ulm, Germany.
  • Soller K; Institute of Molecular Medicine, Ulm University, 89081 Ulm, Germany.
  • Milsom MD; Division of Experimental Hematology, German Cancer Research Center (DKFZ) and Heidelberg Institute for Stem Cell Technology and Experimental Medicine (HI-STEM), Heidelberg, Germany.
  • Florian MC; 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; C
  • Geiger H; Institute of Molecular Medicine, Ulm University, 89081 Ulm, Germany. Electronic address: hartmut.geiger@uni-ilm.de.
STAR Protoc ; 3(3): 101483, 2022 09 16.
Article in En | MEDLINE | ID: mdl-35769923
ABSTRACT
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).
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Stem Cells / Imaging, Three-Dimensional Limits: Animals Language: En Journal: STAR Protoc Year: 2022 Document type: Article
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Stem Cells / Imaging, Three-Dimensional Limits: Animals Language: En Journal: STAR Protoc Year: 2022 Document type: Article