Droplet Barcoding-Based Single Cell Transcriptomics of Adult Mammalian Tissues.


Journal

Journal of visualized experiments : JoVE
ISSN: 1940-087X
Titre abrégé: J Vis Exp
Pays: United States
ID NLM: 101313252

Informations de publication

Date de publication:
10 01 2019
Historique:
entrez: 29 1 2019
pubmed: 29 1 2019
medline: 23 1 2020
Statut: epublish

Résumé

The analysis of single cell gene expression across thousands of individual cells within a tissue or microenvironment is a valuable tool for identifying cell composition, discrimination of functional states, and molecular pathways underlying observed tissue functions and animal behaviors. However, the isolation of intact, healthy single cells from adult mammalian tissues for subsequent downstream single cell molecular analysis can be challenging. This protocol describes the general processes and quality control checks necessary to obtain high-quality adult single cell preparations from the nervous system or skin that enabled subsequent unbiased single cell RNA sequencing and analysis. Guidelines for downstream bioinformatic analysis are also provided.

Identifiants

pubmed: 30688305
doi: 10.3791/58709
doi:

Types de publication

Journal Article Research Support, Non-U.S. Gov't Video-Audio Media

Langues

eng

Sous-ensembles de citation

IM

Subventions

Organisme : CIHR
Pays : Canada

Auteurs

Jo Anne Stratton (JA)

Hotchkiss Brain Institute, University of Calgary; Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary; Alberta Children's Hospital Research Institute, University of Calgary.

Sarthak Sinha (S)

Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary.

Wisoo Shin (W)

Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary.

Elodie Labit (E)

Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary.

Tak-Ho Chu (TH)

Hotchkiss Brain Institute, University of Calgary; Department of Clinical Neurosciences, Cumming School of Medicine, University of Calgary.

Prajay T Shah (PT)

Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary.

Rajiv Midha (R)

Hotchkiss Brain Institute, University of Calgary; Department of Clinical Neurosciences, Cumming School of Medicine, University of Calgary.

Jeff Biernaskie (J)

Hotchkiss Brain Institute, University of Calgary; Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary; Alberta Children's Hospital Research Institute, University of Calgary; jabierna@ucalgary.ca.

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Classifications MeSH