Cell Cycle Analysis
Cell cycle analysis is a very common application of Flow Cytometry. By using a DNA-specific stain one can determine the DNA profile e.g. find percentage of the population in G0/G1, S, and G2/M. This information can be used for example to monitor the effect of the anticancer treatment.
The Flow Cytometry now offers an online Multicycle AV addon to our FCS Express analysis program. Please contact the facility if you need help using Multicycle.
Below are some examples of cell cycle data acquired using a BD LSR II flow cytometer with FCS Express software analysis and this same data using mathematical modeling using ModFif software.
FCS Express Analysis
Diploid |
100% |
Diploid G1 |
82% |
Diploid G2 |
6.72% |
Diploid S |
11.28% |
G2/G1 |
1.90 |
Modfit Analysis
(with singlet discrimination)
Diploid |
100% |
Diploid G1 |
79.77% |
Diploid G2 |
4.57% |
Diploid S |
15.65% |
G2/G1 |
1.92 |
Common Cell Cycle Dyes Used at Facility
Probes |
Facility Instruments |
Excitation laser line |
Emission Filter [nm] |
Propidium Iodide
{Dead Cells} |
BD LSR II
BD FACSCanto
Coulter Epics XL
MoFlo |
488 nm
488 nm
488 nm
488 nm |
695/40
670 LP
630 BP
630/40 |
Hoechst
{Live Cells} |
BD LSR II
MoFlo |
407 nm
351 nm |
450/50
450/65 |
7 Aminoactinomycin D (7AAD)
{Dead Cells} |
BD LSR II
BD FACSCanto
Coulter Epics XL
MoFlo |
488 nm
488 nm
488 nm
530 nm |
695/40
670 LP
630 BP
630/40 |
DAPI
{Dead Cells} |
BD LSR II
MoFlo |
407 nm
351 nm |
450/50
450/65 |
DRAQ5
{Live Cells} |
MoFlo |
647 nm |
675 /30 |
| Flow Cytometry Facility
Barbara Pilas, Ph.D - Director
231 Edward R. Madigan Laboratory, 1201 W. Gregory Drive, Urbana, IL 61801
Phone: (217) 244-0559 FAX: (217) 244-0446 Email: http://illinois.edu/
|
| Last edited:05 June
2007
|
