Laser Scanning Slide & Microplate Cytometry cell cycle analysis on LSC
DRAQ5™ has stoichiometric binding to DNA...
Advantages of using DRAQ5™ in the laser scanning cytometer:-
- "trigger" for LSC event analysis
- cell cycle metric on LIVE or fixed cells: no RNase or UV source required
- less spectral overlap with the green channel e.g. FITC, EGFP, Alexa 488
Reference
DRAQ5™ has been the nuclear dye of choice in a study on the effects of a drug (geldanamycin) treatment and hypoxia on two cancer cell lines using a laser scanning cytometer. (Schwock et al, 2005).
Cells were monitored for internal accumulation of a key signalling molecule (HIF-1α) with Alexa 488 labelled antibody and for impact on cell cycle using the DRAQ5™ nuclear signal.
DRAQ5™ was selected as the preferred "trigger" for cell events and cell cycle (cf. PI) as it has minimal spectral overlap into Alexa 488 (fluorochrome for secondary antibody), does not require RNase treatment and can be excited at 633 nm for DNA measurement in a separate scan acquisition from Alexa 488.
See a review of the Schwock et al. methodology here.
Schwock J, Geddie WR, Hedley DW. Analysis of Hypoxia-Inducible Factor-1α Accumulation and Cell Cycle in Geldanamycin-Treated Human Cervical Carcinoma Cells by Laser Scanning Cytometry. Cytometry Part A 2005; 68A:59-70.
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