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Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids
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Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids

Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids

DOI:
10.3791/56662-v

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09:51 min

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December 13, 2017

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, , , ,
1School of Biological Sciences,University of East Anglia, 2Norwich Medical School,University of East Anglia

Chapters

  • 00:05Title
  • 01:41Isolation of Intestinal Organoids from Basement Matrix Domes in 24-well Plates
  • 03:20Fixation of Isolated Organoids
  • 04:22Blocking, and Primary and Secondary Antibody Incubation
  • 06:41Mounting Isolated Organoids
  • 07:39Results: Isolation and Immunolabeling of Intestinal Organoids
  • 08:55Conclusion

Summary

자동 번역

자동 번역

We present protocols for isolation of intestinal 3D structures from in vivo tissue and in vitro basement matrix embedded organoids, and detail different fixation and staining protocols optimized for immuno-labeling of microtubule, centrosomal, and junctional proteins as well as cell markers including the stem cell protein Lgr5.

Tags

Keywords: 3D Intestinal OrganoidsImmunofluorescent LabelingMicrotubulesCentrosomal ProteinsFixationIsolationImmunostaining

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