Cell Sorting

Cell Sorting

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  • Cell Sorting: Fundamentals and Selected Novel Applications
  • Cell Sorting for Function and Viability
  • 3D Cytometry: Intelligent Image—Activated Cell Sorting
  • Current Standards in Flow Cytometry Cell Sorter Biosafety
  • Intelligent Image-Activated Cell Sorting: A Tutorial 
  • Optimizing SRL Performance: Boost your Cell Sorting Capacity
  • Optimizing SRL Performance: Boost your Cell Sorting Capacity

    Contains 4 Component(s), Includes Credits Recorded On: 10/29/2015

    CYTO U Webinar presented by Rui Gardner, PhD Keywords: Instrument Management, Cell Sorting, Staff Management, Self-Operation, Training

    About the Presenter

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    Rui Gardner, PhD
    Head of the Flow Cytometry Core Facility
    Memorial Sloan Kettering Cancer Center

    Rui Gardner’s scientific career began just before graduating with a BS in biochemistry in 1997, as a trainee in mathematical biology. This set the ground for his doctoral work, a mathematical approach to understand the prooxidative mechanisms of superoxide dismutase, paradoxically known as one of the most powerful physiological antioxidant enzymes. Most of the work was carried out in the Department of Microbiology and Immunology at the University of Michigan but also at the University of Southern California and the Gulbenkian Institute of Science in Portugal. In 2004, Rui earned his PhD in biomedical sciences, followed by postdoctoral work on evaluating immune diversity estimation techniques. He became SRL Manager of the Flow Cytometry facility in 2006 at the Gulbenkian Institute of Science. Since 2007, Rui has been actively involved with ISAC’s SRL Task Force. In 2012 he was elected for ISAC’s Council, currently chairing the ISAC SRL Oversight Committee responsible for implementing and improving ISAC’s SRL-related activities and programs. Over the past several years, Rui has focused his efforts in addressing cell sorting performance based on the almost 10-year long experience running a cell sorting facility, as well as promoting meetings and discussions within the SRL community to help improve facility operations and management.  

    Webinar Summary

    In the first of a series of webinars addressing strategies to optimize operations and management in a Shared Resource Lab (SRL), we’ll focus on enhancing cell sorting capacity. Many sorting facilities struggle with the increased demand in usage. The typically limited installed capacity of droplet cell sorters, which is directly associated to the requirement of a dedicated operator, usually leads to a narrow range of solutions to increase this capacity. Strategies are most often reduced to buying a new instrument, implying an increase in the number of FTEs, or most likely increasing dramatically the burden of the current FTE, inevitably decreasing quality or SRL performance. With the advent of the new and more automated cell sorters, different and more creative solutions to increase cell sorting capacity are being addressed, including self-service sorting with all the implications associated to it. We’ll discuss several of these strategies, and present additional approaches to optimize usage of the current instrumentation, independent of their level of automation. These may include small technical implementations to reduce instrument setup, instrument troubleshooting guides, booking strategies to optimize resource usage, staggering shifts, or “on call” staff to assist sorts that cannot be scheduled within normal sorting hours. There is no general strategy that will fit all facilities as some of these approaches may even collide with institutional or country-specific policies. Nevertheless, our aim is to lay down some of the possible strategies, open a discussion towards addressing this issue in a systematic way, and hopefully inspire new creative solutions from peers.

    CMLE Credit: 1.0

  • Current Standards in Flow Cytometry Cell Sorter Biosafety

    Contains 3 Component(s), Includes Credits Recorded On: 06/22/2019

    A CYTO 2019 Scientific Tutorial Presented by Geoffrey Lyon, Steve Perfetto, & Evan Jellison Keywords: aerosol testing, risk assessment, Glo-Germ testing, UV-aerodynamic particle sizer, YG beads, Cyclex-D test, Dragon Green Beads, aerosol evacuation system, SOPs

    The Presenters

    Geoffrey Lyon
    Yale University School of Medicine

    Steve Perfetto
    NIAID, National Institute of Health

    Evan Jellison
    UCONN Health

    Session Summary

    This tutorial will provide a summary of biosafety principles as they apply to flow cytometry and cell sorting, including a comprehensive review of an improved bead-based containment assay presented in paper “Novel Impactor and Microsphere-Based Assay Used to Measure Containment of Aerosols Generated in a Flow Cytometer Cell Sorter” (Perfetto. S., Cytometry A 2019; 95A: 173-182). This tutorial will also provide an expert forum highlighting specific scenarios that operators or core facility managers encounter. After this tutorial, the attendee should have a clearer understanding of the principles and practices of biosafety as they pertain to flow cytometry, in particular cell sorting. Additionally, the attendees will have an understanding of how to properly conduct this new bead-based assay to validate the containment of cell sorters.

    CMLE Credit: 1.5

  • Cell Sorting for Function and Viability

    Contains 3 Component(s), Includes Credits Recorded On: 04/28/2018

    A CYTO 2018 Scientific Tutorial presented by Monica DeLay, Peter Lopez, and Matthias Schiemann Keywords: History, Sorter-Induced Cellular Stress (SICS), sorting conditions, mechanical forces, Sheath buffer, sample buffers, nozzle, pressure, temperature, Instrument type, Sorting time, troubleshoot

    The Presenters

    Monica DeLay
    Cincinnati Children's Hospital
    USA

    Peter Lopez
    NYU School of Medicine
    USA

    Matthias Schiemann
    Technical University of Munich
    Germany

    Session Summary

    The objective of a successful sort is to obtain viable cells functionally representative of the unmanipulated target population. Toward this goal, there are a number of factors to consider depending on cell type and downstream application. This tutorial will review basic guidelines (e.g., buffers, nozzle size, and pressure) that are broadly applicable for obtaining useful sorted product. Specific examples where conditions were optimized for certain cell types will be highlighted. Methods to ascertain cell perturbation as a result of sorting will also be discussed. In addition, attendees are encouraged to bring their relevant experiences to share.

    CMLE Credit: 1.5

  • Cell Sorting: Fundamentals and Selected Novel Applications

    Contains 3 Component(s), Includes Credits Recorded On: 06/11/2016

    A CYTO 2016 Scientific Tutorial presented by Joe Trotter, Geoffrey Osborne, and Charles "Hank" Pletcher Keywords: Decision deflection, droplet break-off, nozzle, purity, recovery, sample quality, sample preparation, sample buffer, index sort, pulse geometry, singlets, duplets, fanning, accuracy drop, sorting troubleshooting

    The Presenters

    Joe Trotter
    Principal Scientist
    BD Fellow

    Geoffrey Osborne, PhD
    Director of Flow Cytometry
    University of Queensland

    Charles "Hank" Pletcher

    Session Summary

    This session will provide the attendee with theoretical and practical understanding applicable to the various cell separation scenarios that are common to most current laboratory needs, workflows, and protocols.

    CMLE Credit: 1.5