MENU
FEB 21, 2018 6:00 AM PST

Multiparameter Cell Cycle Analysis

Speaker
  • Professor Emeritus (Oncology), Director Cytometry & Microscopy Core, Case Comprehensive Cancer Center, Co-Director Immune Function Core, Case Center for AIDS Research
    Biography
      James Jacobberger earned degrees in Microbiology and Immunology (BS, University of Nebraska (1977), MS, PhD, University of Rochester (1983)) and studied Experimental Pathology as a Post Doctoral Fellow (Dept. Pathology, University of Colorado (1983-1985). His PhD thesis and Post-Doctoral work emphasized concentrated efforts to apply cytometry to malaria and oncologenic virology problems. He was appointed Assistant Professor of Anatomy and Developmental Genetics (1985) at Case Western Reserve University and promoted to Associate Professor of General Medical Sciences (GMS-Oncology) with Tenure (1993), Professor of GMS-Oncology (2001), Professor Emeritus (2016). Dr. Jacobberger served as Associated Director for Shared Resources of the Case CCCC from 2006 - 2016. He created the current Case Comprehensive Cancer Center's Cytometry and Microscopy Shared Resource (CMSR) in 1985 with a single cell sorter. This (now) large facility thrives today with 4 Associate Directors, a staff of 3, and provides comprehensive cytometry and microscopy services to >100 laboratories per year. It is NIH funded as part of the Case Comprehensive Cancer Center and Case Center for AIDS Research. Dr. Jacobberger's research interests focus on cytometry with biological interests in cell cycle, oncogenesis, and biomarkers of clinical relevance in cancer. His research laboratory was continuously NIH funded for ~30 years. He has been an Associate Editor of the journal, Cytometry A. He has served on many NIH and other study sections, generally in the areas of instrumentation, biotechnology, and biomarkers. In April, 2018, Dr. Jacobberger will step down as Director of the CMSR but continue as part of the research services part of the CMSR. His future goals are to continue research and teaching.

    Abstract

    Most cell cycle assays amount to counting cells and/or measuring DNA content and deconvolving the resulting histogram into G1, S, G2+M, or G1, S, G2, and M.  Considerable effort has been put to correlating DNA content with RNA, protein, or specific epitopes to determine or discover additional compartments.  Here, we'll focus on flow cytometric analysis of DNA and specific epitopes.  In recent efforts, using DNA peak to mark anaphase/cytokinetic populations in a 6 parameter analysis (DNA-area, DNA-peak, Light Scatter, cyclin A2, cyclin B1, and phospho-S10-histone H3), we identify 15 compartments with more than 4 additional compartments representing apoptotic cells.

    Rational approaches to fixation and optimized staining protocols were worked out years ago.  We have examined many experimental variations on the basic protocols without uncovering any major improvements that result in higher quality data.  We are working currently on a “washless” staining assay, relying on a single high dilution to minimize background staining and relying on acoustic focusing to render the sample analyzable over a short period.  The results are striking. The S/N is equivalent to a fully washed sample; the minimized handling results in better cell recovery, and improved recovery results in better definition of cell cycle compartments with low cell numbers.  Additionally, the “washless” assay provides significant labor savings.  For research use only.  Not for use in diagnostic procedures.

    Learning Objectives:

    • overview of fixation & staining for intracellular epitopes
    • underlying logic and purpose of cell cycle analysis
    • in depth, analytical protocol for multiparameter analysis backbone

    Show Resources
    You May Also Like
    MAY 11, 2021 10:00 AM PDT
    Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
    C.E. CREDITS
    MAY 11, 2021 10:00 AM PDT
    Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
    Date: May 11, 2021 Time: 10:00zm PDT Your samples are some of the most valuable assets in the laboratory. After spending countless hours on extraction and preparation, your conclusions could...
    JUN 09, 2021 7:00 AM PDT
    Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
    C.E. CREDITS
    JUN 09, 2021 7:00 AM PDT
    Add to Calendar Select one of the following: iCal Google Calendar Outlook Calendar Yahoo Calendar
    Date: June 9, 2021 Time: 09 June 2021, 7am PDT, 10am EDT, 4pm CEST cells with dramatic implications on the validity of past cell culture related research. The fact that at least 509 cell lin...
    OCT 29, 2020 6:00 AM PDT
    C.E. CREDITS
    OCT 29, 2020 6:00 AM PDT
    Date: October 29, 2020 Time: 6:00am (PDT), 9:00am (EDT), Chronic inflammation can occur as a result of a combination of genetic predispositions and environmental factors. Epigenetic modifica...
    NOV 16, 2020 8:00 AM PST
    C.E. CREDITS
    NOV 16, 2020 8:00 AM PST
    Date: November 16, 2020 Time: 8:00am (PST), 11:00am (EST) CRISPR screening has become the prime discovery tool in modern biomedical research and drug discovery. At the same time, most screen...
    DEC 02, 2020 8:00 AM PST
    C.E. CREDITS
    DEC 02, 2020 8:00 AM PST
    DATE: December 2nd, 2020 TIME: 08:00am PDT, 11:00pm EDT Bioreactors and shakers are used to cultivate microorganisms, plant, insect, and mammalian cells in different volumes. Upscaling of pr...
    MAR 16, 2021 10:00 AM PDT
    C.E. CREDITS
    MAR 16, 2021 10:00 AM PDT
    Date: March 16, 2021 Time: 10:00am (PST) Scientific progress and breakthroughs today are often too expensive for most institutions to acquire. Each year, the National Institutes of Health (N...
    FEB 21, 2018 6:00 AM PST

    Multiparameter Cell Cycle Analysis


    Specialty

    Cell Biology

    Cancer Research

    Cell Culture

    Immunology

    Molecular Biology

    Biotechnology

    Cell Signaling / Transduction

    Cancer Therapeutics

    Cancer Diagnostics

    Biomarkers

    Antibodies

    Gene Expression

    Biochemistry

    Genetics

    Immuno-Oncology

    Geography

    North America52%

    Europe24%

    South America11%

    Asia7%

    Africa2%

    Oceania1%

    Registration Source

    Website Visitors79%

    WAVE Newsletter Promotion10%

    Email Promotions7%

    Social Media Promotions3%

    Website Banners1%

    Client/Sponsor Promotions1%

    Job Title

    Student21%

    Research Scientist17%

    Scientist9%

    Educator/Faculty8%

    Facility/Department Manager8%

    Executive7%

    Post Doc7%

    Medical Doctor/Specialist6%

    Medical Laboratory Technician6%

    Lab Management5%

    Biologist3%

    Clinical Laboratory Scientist2%

    Veterinarian1%

    Organization

    Academic Institution19%

    Research Institute12%

    Hospital6%

    Clinical Laboratory5%

    Biotech Company5%

    Government4%

    Life Science Company3%

    Medical School3%

    Medical Center3%

    Pharmaceutical Company2%

    Contract Research Organization (Cro)1%

    Ambulatory Care1%

    Industrial Company1%

    Non-Profit Organization1%

    Other5%


    Show Resources
    Loading Comments...
    Show Resources
    Attendees
    • See more