Preparing Mouse Organs for Flow Cytometry

 
LEAD AUTHOR

Jennifer Kay, Graduate student, MIT Department of Biological Engineering, Cambridge, MA, USA

jekay@mit.edu

OTHER AUTHORS:

Dr. Michelle Sukup-Jackson, former graduate student
Dr. Orsolya Kiraly, former post-doc
Dr. Bevin Engelward, Professor
All from MIT Department of Biological Engineering, Cambridge, MA


PROTOCOL TYPE: Real Methods
Protocol Document
Click here for example data file.
SYNOPSIS:

This protocol describes how to disaggregate fresh tissues into single cell suspensions for analysis by flow cytometry. The protocol was developed for analyzing the proportion of EGFP+ cells in RaDR mouse tissues, but it can be adapted to any application requiring a suspension of single cells.


ABSTRACT:

This protocol is a description of the Real Methods used to disaggregate tissues into single cell suspensions. In order to perform flow cytometric analysis on single cells from mice, it is necessary to mechanically mince the tissue and chemically digest connective components while leaving cells alive and intact. This protocol provides information to create cell suspensions from pancreas, liver, breast, intestine, spleen, brain, kidney, thymus, and lung tissues.


 
PUBLICATIONS:

Kiraly O, Gong G, Olipitz W, Muthupalani S, Engelward BP (2015) Inflammation-Induced Cell Proliferation Potentiates DNA Damage-Induced Mutations In Vivo.PLoS Genet 11(2): e1004901. doi: 10.1371/journal.pgen.1004901

Sukup-Jackson MR, Kiraly O, Kay JE, Na L, Rowland EA, et al. (2014) Rosa26-GFP Direct Repeat (RaDR-GFP) Mice Reveal Tissue- and Age-Dependence of Homologous Recombination in Mammals In Vivo. PLoS Genet 10(6): e1004299. doi: 10.1371/journal.pgen.1004299


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