Image - Barts and The London logo and link to home page Image - divider Image - divider
 
  Institute of Cell and Molecular Science
Flow Cytometry Core Facility equipment, images used with permission by BD
  Flow Cytometry Core Facility
  link Home link Instruments available link Flow cytometry link Uses of flow cytometry link Flow sorting link Links link Contacts
 

Cell cycle determination by epi-fluorescence microscopy

 

The determination of the cell cycle analysis is a well known technique in flow cytometry. The Core Facilities at ICMS has a Metamorph epi-fluorescent deconvolution system with a software macro to determine the cell cycle stage of cells on slides or in culture.

Jurkat cells stained with DNA binding dyes such as propidium iodide (PI) or DAPI can be analysed by Metamorph to determine the percentage of cells in G1, S, G2m and apoptosis.

Jurkat cells were sorted on a BD FACSAria into G1, S phase and G2m cells. Centrifuged cells were placed onto microscope slides and imaged at the same exposure and magnification (x20). This allows the system to be calibrated so that unsorted cells can be analysed using the Metamorph cell cycle analysis macro.

 
Top
 

PI cell cycle sorting

 

G1 calibration

 

Cell cycle image analysis

by Gary Warnes. © Queen Mary, University of London 2007
Institute of Cell and Molecular Science, The Blizard Building, Barts and The London School of Medicine and Dentistry, 4 Newark Street, London E1 2AT, UK Tel: +44 (0)20 7882 2483, Fax: +44 (0)20 7882 2200