Quantifying Heterogeneity of Individual Organelles in Mixed Populations via Mass Cytometry

Heather M.G. Brown, Edgar A. Arriaga

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Macroautophagy is a complex degradative intracellular process by which long-lived proteins and damaged organelles are cleared. Common methods for the analysis of autophagy are bulk measurements which mask organelle heterogeneity and complicate the analysis of interorganelle association and trafficking. Thus, methods for individual organelle quantification are needed to address these deficiencies. Current techniques for quantifying individual autophagy organelles are either low through-put or are dimensionally limited. We make use of the multiparametric capability of mass cytometry to investigate phenotypic heterogeneity in autophagy-related organelle types that have been isolated from murine brain, liver, and skeletal muscle. Detection and phenotypic classification of individual organelles were accomplished through the use of a lanthanide-chelating membrane stain and organelle-specific antibodies. Posthoc sample matrix background correction and nonspecific antibody binding corrections provide measures of interorganelle associations and heterogeneity. This is the first demonstration of multiparametric individual organelle analysis via mass cytometry. The method described here illustrates the potential for further investigation of the inherently complex interorganelle associations, trafficking, and heterogeneity present in most eukaryotic biological systems.

Original languageEnglish (US)
Pages (from-to)13315-13321
Number of pages7
JournalAnalytical Chemistry
Volume90
Issue number22
DOIs
StatePublished - Nov 20 2018

Bibliographical note

Funding Information:
We thank Nick Livezey for assistance metallating the didodecyl-DTPA reagent and interpretation of the 1H NMR of the nonmetalated ligand. H.M.G.B. and E.A.A. thank the National Institutes of Health (Grants RO1-AG020866 and T32-GM008347).

Publisher Copyright:
© 2018 American Chemical Society.

Fingerprint

Dive into the research topics of 'Quantifying Heterogeneity of Individual Organelles in Mixed Populations via Mass Cytometry'. Together they form a unique fingerprint.

Cite this