NOV 19, 2020 8:00 AM GMT

Highly specific multiplexed RNA imaging in tissues with split-FISH

Sponsored by: Andor
C.E. Credits: P.A.C.E. CE Florida CE
Speakers
  • Senior Research Scientist, Genome Institute of Singapore
    Biography

      Kok Hao is currently a senior research scientist at the Genome Institute of Singapore (GIS). Prior to his current appointment, Kok Hao was a GIS Fellow and AXA Postdoctoral Fellow at the GIS between 2016 and 2019. From 2009 to 2015, he completed his Ph.D. with Xiaowei Zhuang group at the Department of Chemistry and Chemical Biology, Harvard University. Kok Hao graduated with a Bachelor of Science from the Department of Chemical and Biomolecular Engineering with a minor in Physics at the University of Illinois at Urbana Champaign. He is a recipeint of the AXA Postdoctoral Fellowship Award in 2016, NMRC Young Investigator Award in 2015, and A*STAR National Science Scholarship in 2009.

    • Product Specialist for Microscopy Cameras at Andor Technology.
      Biography
        My background in microscopy is built from my work and studies while at Queen's University Belfast. I always had a keen interest in bacteria and viruses as pathogens, as well as how we may exploit them for our benefit, so naturally specialised in microbiology. After my BSc (Hons) in 1999, I went on to do my PhD in microbiology within the School of Biology and Biochemistry at the Medical Biology Centre at Queens. My PhD was focused on polyphosphate metabolism of microorganisms- trying to get a better understanding of this molecule, how to quantify it, and the key roles this played within environmental and pathogenic bacteria. Roles include as a protective function in response to stress and in helping pathogens establish infections. An interesting application of this was in biological phosphate removal from wastewater for which we had a pilot scale study. This then led on to a Research Fellow position in the same research group of John Quinn and John McGrath in 2002. While in this position I worked further on connecting the biochemistry and genetics of polyphosphate metabolism. This included studies to characterise the bacterial transport systems involved and development of novel research tools, such as fluorescent assays to determine intracellular pH, and how to apply our luminescence and fluorescence imaging based methods to 96 well plate formats for faster screening and analysis. After Academia, I went on to work in the Medical Diagnostics and Pharmaceutical industries. I joined Andor Technology in 2012, initially working as a Technical Author for our camera and systems products. My current role is a Product Specialist for microscopy cameras. This role brings me in touch with the latest developments in camera technology, as well as a broad range of microscopy applications and techniques.

      Abstract
      Date:  November 19, 2020
      Time:  8:00am (GMT),  9:00am (CET),  4:00pm (CST)
       
      We present split-FISH, a multiplexed fluorescence in situ hybridization method that leverages a split-probe design to achieve enhanced specificity. Split-FISH reduces off-target background fluorescence, decreases false positives and enables accurate RNA profiling in uncleared tissues. We demonstrate the efficacy of split-FISH on various mouse tissues by quantifying the distribution and abundance of 317 genes in single cells and reveal diverse localization patterns for spatial regulation of the transcriptome in complex tissues.
       
      Learning Objectives
      • Multiplexed FISH is an attractive tool for spatial transcriptomics.
      • Advantages of split-FISH design in lowering background and false positive noise
      • Example applications in profiling RNA localizations in mouse organs
       
       
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