NOV 17, 2016 08:30 AM PST

Simultaneous, single-cell visualization of RNA and Protein

  • Associate Director R&D, eBioscience/Affymetrix, now a part of Thermo Fisher Scientific
      Dr. Sara Becker-Catania is an Associate Director in R&D at eBioscience and received her PhD in Experimental Pathology at University of California, Los Angeles studying the genotype/phenotype correlations of a rare neurodegenerative disorder, ataxia telangiectasia. She did her postdoctoral training as a Giannini Foundation fellow at the UCLA Neuropsychiatric Institute/Mental Retardation Research Center studying the selected specification of neural stem cells along neural and glial lineages. Following her postdoctoral training, she held a joint appointment as a Research Assistant Professor at University of Illinois Chicago and the Hines VA Hospital and was a recipient of an Illinois Regenerative Medicine Institute stem cell grant for her work with neural stem cells in models of Multiple Sclerosis. She joined eBioscience, establishing a group developing products for microscopy and imaging applications. Currently, she leads a team dedicated to the development of immunology-focused reagents for flow cytometry and microscopy applications.

    DATE: November 17, 2016
    TIME: 8:30am PT, 11:30am ET

    The complex interactions between transcription, translation, and post-translational modifications are hidden from view in typical endpoint assays that measure either RNA or protein levels. While in situ hybridization (ISH) provides a method for examining the levels of specific RNA transcripts in individual cells, and immunocytochemistry (ICC) utilizes antibodies to visualize the localization of specific proteins (and protein modifications), the ability to simultaneously observe RNA and protein in a single cell has been thwarted by the incompatibility of ICC and ISH protocols. Further complicating this analysis, traditional ISH methods lack sensitivity for detecting low-abundance RNA species and have limited multiplexing capability that further constrain its compatibility with other cell assays.

    This webinar will describe a novel technique combining ICC staining with RNA ISH, utilizing branched DNA signal amplification technology, allowing for single copy mRNA sensitivity. We will demonstrate the utility and versatility of this technique using a variety of antibody and probe combinations relevant for cancer research. We will compare expression of protein and mRNA of HER2, estrogen receptor alpha, and progesterone receptor in several breast cancer cell lines known to express differential levels of these targets. We will also compare expression of several microRNAs known to play a role in cancer development in combination with staining for proteins involved in proliferation, anti-apoptosis, and cell structure. Finally, we will show how this novel technique can be used for other research applications for example the detection of Zika virus RNA in infected cells, analysis of circulating tumor cell models and phenotyping of cells within primary hippocampal cultures.

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