Simultaneous Proteomics and Transcriptomics TotalSeq™ and The Future of Single Cell Analysis
Miguel A. Tam
Thursday, May 16, 2019
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Chapters
Introduction
Overview
Why analyzing RNA in single cells?
Droplet-based scRNA-seq profile thousands of cells in parallel
RNA and proteins expression doesn’t always correlate
Proteomic technologies are lagging in the era of NGS
Simultaneous RNA and protein analysis
Protein detection using NGS as readout
Protein abundance readout using tagged antibodies
CITE-seq workflow and TotalSeq™
Overview
Integrated solutions for every experimental design
Integrated solutions for every experimental design – Cell Hashing
BioLegend Cell Hashing reagents
Overview
Cell Hashing recovers expected cell proportions
Samples identified with hashtags
Memory B cell differentiation in the context of a novel influenza vaccine
Differential marker expression using scRNA-seq
Expansion with TotalSeq™-C
Identification of unique receptor expression
Experimental design
Clustering Maps
Clustering Results
Full cluster expression results
Identification of unique markers
Overview
Clustering PBMCs in 220-plex Identifies CD8+ T-Cell Subsets
Clustering PBMCs via 9-plex TBNK identifies major immune cell types
Intracellular staining – ZAP-70
Conclusions
Acknowledgements
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