SEP 15, 2015 07:00 AM PDT

WEBINAR: Less False Negatives: Quantifying Cell Viability by Simultaneous Triple Staining

Speakers
  • Research Scientist, Kroemer Lab, Paris, France
    Biography
      Oliver Kepp received his Ph.D. in 2006 from the Humboldt University of Berlin and the Max Planck Institute for Infection Biology in Berlin, Germany. He is currently a research scientist in the laboratory of Guido Kroemer, where he investigates several aspects of immunogenic cell death, focusing on systems biology approaches.
    • Research Engineer, Institut Gustave Roussy, Villejuif, France, Kroemer Lab, Paris, France
      Biography
        Allan Sauvat received his engineer's degree in 2011 from the Institut Supérieur de BioSciences in Paris, France. He is now a research engineer on the BioCell platform in Villejuif, France, where is focusing on HTS assay and software development for systems biology.

      Abstract:
      Cellular viability is usually determined by measuring the capacity of cells to exclude vital dyes such as 4',6-diamidino-2-phenylindole (DAPI), or by assessing nuclear morphology with chromatinophilic plasma membrane-permeant dyes, such as Hoechst 33342. However, a fraction of cells that exclude DAPI or exhibit normal nuclear morphology have already lost mitochondrial functions and/or manifest massive activation of apoptotic caspases, and hence are irremediably committed to death. Here, we developed a protocol for the simultaneous detection of plasma membrane integrity (based on DAPI) or nuclear morphology (based on Hoechst 33342), mitochondrial functions (based on the mitochondrial transmembrane potential probe DiOC6(3)) and caspase activation (based on YO-PRO®-3, which can enter cells exclusively upon the caspase-mediated activation of pannexin 1 channels). This method, which allows for the precise quantification of dead, dying and healthy cells, can be implemented on epifluorescence microscopy or flow cytometry platforms and is compatible with a robotized, high-throughput workflow.

      Learning Objectives:
      • Learn about the protocol developed for the simultaneous detection of plasma membrane integrity (based on DAPI) or nuclear morphology (based on Hoechst 33342), mitochondrial functions (based on the mitochondrial transmembrane potential probe DiOC6(3)) and caspase activation (based on YO-PRO®-3, which can enter cells exclusively upon the caspase-mediated activation of pannexin 1 channels)
      • Learn how this method, which allows for the precise quantification of dead, dying and healthy cells, can be implemented on epifluorescence microscopy or flow cytometry platforms and is compatible with a robotized, high-throughput workflow

      Show Resources
      You May Also Like
      MAY 24, 2018 09:30 AM PDT
      C.E. CREDITS
      MAY 24, 2018 09:30 AM PDT
      DATE: May 24, 2018 TIME: 9:30PM PDT The current gold standard in in vitro pre-clinical cancer treatment screening remain cell lines,...
      JUN 26, 2018 06:00 AM PDT
      C.E. CREDITS
      JUN 26, 2018 06:00 AM PDT
      Date: June 26, 2018Time: 6:00 a.m. PDT, 9:00 a.m. EDT, 1500 CEST Today’s hematology analyzers employ various methods for enumerating platelets. These methods include: e...
      MAY 03, 2018 11:00 AM PDT
      MAY 03, 2018 11:00 AM PDT
      DATE: May 3, 2018TIME: 11:00AM PDT, 2:00PM EDTWhile stress is one of the leading causes of neuropsychiatric disorders, the molecular underpinnings of how stress induces alterations in b...
      APR 27, 2018 10:00 AM PDT
      C.E. CREDITS
      APR 27, 2018 10:00 AM PDT
      DATE: April 27, 2018TIME: 10:00am PST, 1:00pm ESTGlioblastoma (GBM) and Medulloblastoma (MB) are the most common adult and paediatric brain tumours, both of which can have devastating c...
      JUN 20, 2018 10:00 AM EDT
      C.E. CREDITS
      JUN 20, 2018 10:00 AM EDT
      DATE: June 20, 2018TIME: 07:00AM PDT, 10:00AM EDTIntroducing GE’s New Lyo-StableTM service. Sepsis is one of the top challenges facing hospitals in terms of clinical outcomes...
      MAY 22, 2018 08:00 AM PDT
      C.E. CREDITS
      MAY 22, 2018 08:00 AM PDT
      DATE: May 22, 2018TIME: 08:00AM PDT The nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) are closely related transcription factors that...
      Loading Comments...