Highly Multiplexed, Single Cell Transcriptomic Analysis of T-Cells by Microfluidic PCR
- PMID: 27787802
- DOI: 10.1007/978-1-4939-6548-9_15
Highly Multiplexed, Single Cell Transcriptomic Analysis of T-Cells by Microfluidic PCR
Abstract
Recently, technologies have been developed to measure expression of 96 (or more) mRNA transcripts at once from a single cell. Here we describe methods and important considerations for use of Fluidigm's BioMark platform for multiplexed single cell gene expression. We describe how to qualify primer/probes, select genes to examine in 96-parameter panels, perform the reverse transcription/cDNA synthesis step, and operate the instrument. In addition, we describe data analysis considerations. This technology has enormous value for characterizing the heterogeneity of T-cells, thereby providing a useful tool for immune monitoring.
Keywords: Microfluidic; Quantitative competitive real-time PCR; Single cell analysis; T-cell; Transcriptome.
Similar articles
-
Microfluidic single-cell whole-transcriptome sequencing.Proc Natl Acad Sci U S A. 2014 May 13;111(19):7048-53. doi: 10.1073/pnas.1402030111. Epub 2014 Apr 29. Proc Natl Acad Sci U S A. 2014. PMID: 24782542 Free PMC article.
-
Gene Expression Analysis by Multiplex Single-Cell RT-PCR.Methods Mol Biol. 2019;1941:139-154. doi: 10.1007/978-1-4939-9077-1_10. Methods Mol Biol. 2019. PMID: 30707432
-
Quantitative assessment of single-cell RNA-sequencing methods.Nat Methods. 2014 Jan;11(1):41-6. doi: 10.1038/nmeth.2694. Epub 2013 Oct 20. Nat Methods. 2014. PMID: 24141493 Free PMC article.
-
Advancing single-cell proteomics and metabolomics with microfluidic technologies.Analyst. 2019 Jan 28;144(3):846-858. doi: 10.1039/c8an01503a. Analyst. 2019. PMID: 30351310 Review.
-
Detecting Antigen-Specific T Cell Responses: From Bulk Populations to Single Cells.Int J Mol Sci. 2015 Aug 12;16(8):18878-93. doi: 10.3390/ijms160818878. Int J Mol Sci. 2015. PMID: 26274954 Free PMC article. Review.
Cited by
-
Single cell multiomic analysis of T cell exhaustion in vitro.Cytometry A. 2022 Jan;101(1):27-44. doi: 10.1002/cyto.a.24496. Epub 2021 Sep 21. Cytometry A. 2022. PMID: 34390166 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources