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

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
JUN 28, 2022 7:00 AM PDT
JUN 28, 2022 7:00 AM PDT
Date: June 28, 2022 Time: 3:00pm (BST), 4:00pm (CET), 9:00am (CST), 7am (PST) Light-sheet microscopy is an extremely versatile imaging technique with a vast range of implementations that are...
AUG 24, 2022 7:00 AM PDT
AUG 24, 2022 7:00 AM PDT
Date: August 24, 2022 Time: 7:00am (PDT), 10:00pm (EDT), 4:00pm (CEST) Light field microscopy was first introduced in 2006, and allows users to capture the 4D light field within the microsco...
MAY 17, 2022 9:00 AM PDT
MAY 17, 2022 9:00 AM PDT
Date: May 17, 2022 Time: 9:00am (PDT), 12:00pm (EDT), 8:00pm (CEST) Gene therapeutics have great potential to treat many severe diseases in an unprecedented, targeted manner. The biopharmace...
OCT 12, 2022 8:00 AM PDT
OCT 12, 2022 8:00 AM PDT
First Broadcast: Date: October 12, 2022 Time: 8:00am PDT, 11:00am EDT Second Broadcast: Date: October 12, 2022 Time: 9:00am SGT The new Embedded CryoSPARC Live, now fully integrated with t...
AUG 10, 2022 10:00 AM PDT
AUG 10, 2022 10:00 AM PDT
Date: August 10, 2022 Time: 10:00am PDT, 1:00pm EDT The global pandemic has increased focus and scrutiny on molecular diagnostic assay development, resulting in a need for assays that provid...
OCT 11, 2022 8:00 AM PDT
C.E. CREDITS
OCT 11, 2022 8:00 AM PDT
Date: October 11, 2022 Time: 8:00am (PDT), 11:00pm (EDT), 5:00pm (CEST) Multiomic profiling of cell populations at single-cell resolution is revolutionizing scientists’ understanding o...
FEB 21, 2018 6:00 AM PST

Multiparameter Cell Cycle Analysis



Show Resources
Loading Comments...
Show Resources