Journal
/
Developmental Biology
/
Immuno-fluorescent Labeling of Microtubules and Centrosomal Proteins in Ex Vivo Intestinal Tissue and 3D In Vitro Intestinal Organoids
JoVE Journal
Developmental Biology
This content is Free Access.
JoVE Journal Developmental Biology
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

•

09:51 min

•

December 13, 2017

•

, , , ,
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

Automatic Translation

Automatic Translation

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/56662?article_type=v

Viral-mediated Labeling and Transplantation of Medial Ganglionic Eminence (MGE) Cells for In Vivo Studies
check_url/56662?article_type=v

In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition
check_url/56662?article_type=v

Live Imaging of Mouse Secondary Palate Fusion
check_url/56662?article_type=v

Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains
check_url/56662?article_type=v

Processing of Human Cardiac Tissue Toward Extracellular Matrix Self-assembling Hydrogel for In Vitro and In Vivo Applications
check_url/56662?article_type=v

Real-time Measurement of Epithelial Barrier Permeability in Human Intestinal Organoids
check_url/56662?article_type=v

Genetic Engineering of Primary Mouse Intestinal Organoids Using Magnetic Nanoparticle Transduction Viral Vectors for Frozen Sectioning
check_url/56662?article_type=v

Visualizing the Node and Notochordal Plate In Gastrulating Mouse Embryos Using Scanning Electron Microscopy and Whole Mount Immunofluorescence
check_url/56662?article_type=v

Ex Vivo Perfusion of the Rodent Placenta
check_url/56662?article_type=v

Recapitulating Suckling-to-Weaning Transition In Vitro using Fetal Intestinal Organoids

Read Article