Highly multiplexed simultaneous detection of RNAs and proteins in single cells

Nature Methods
Andreas P FreiPier Federico Gherardini

Abstract

To enable the detection of expression signatures specific to individual cells, we developed PLAYR (proximity ligation assay for RNA), a method for highly multiplexed transcript quantification by flow and mass cytometry that is compatible with standard antibody staining. When used with mass cytometry, PLAYR allowed for the simultaneous quantification of more than 40 different mRNAs and proteins. In primary cells, we quantified multiple transcripts, with the identity and functional state of each analyzed cell defined on the basis of the expression of a separate set of transcripts or proteins. By expanding high-throughput deep phenotyping of cells beyond protein epitopes to include RNA expression, PLAYR opens a new avenue for the characterization of cellular metabolism.

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Methods Mentioned

BETA
RNA-seq
flow cytometry
Assay
using flow
fluorescence microscopy
PMA

Software Mentioned

PLAYR
PLAYRDesign
viSNE
Primer3
R

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