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Validating single-cell RNA-seq data using Single-Cell Western blotting and RNAscope® technology
Brian P. Hermann, Ph.D.
Associate Professor, University of Texas, San Antonio
Outline:

This webinar will cover the basics of the Milo Single-Cell Western blot (SCW) and RNAscope single-molecule fluorescence RNA in situ hybridization (smFISH) technologies and present data from an ongoing study in the laboratory of Dr. Brian Hermann at the University of Texas at San Antonio where both platforms are being used to validate single-cell RNA-seq data. Dr. Hermann studies spermatogonial stem cells (SSCs), which are adult stem cells responsible for life-long sperm production in the testes of mammals. Single-cell RNA-seq studies from his laboratory identified substantial gene expression heterogeneity among undifferentiated spermatogonia (which include both SSCs and the progenitor spermatogonia that they produce) at the mRNA level. Results from ongoing validation studies in his laboratory will be presented. Dr. Hermann has used Milo SCW to confirm gene expression heterogeneity at the protein level among individual cells and RNAscope smFISH has validated heterogeneity at the mRNA level and facilitated spatial localization of cell subsets.

Learning Objectives:
  • Understand how Milo Single-Cell Western blotting can provide protein-level validation of heterogeneity among individual cells uncovered by single-cell RNA-seq (scRNA-seq)

  • Understand how RNAscope smFISH can be used to validate scRNA-seq data at the mRNA level and enable spatial remapping to determine localization of cells with particular phenotypes.

 

 

Watch the On-Demand Version Here

 

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