Flow Cytometry cell ploidy / aneuploidy
establish ploidy status in cell populations...
DRAQ5™ can be used directly with FITC & PE antibodies and on intact cells ....
Certain tumour types exhibit aneuploidy i.e. missing or additional chromosomes (hypoploidy or hyperploidy) and this can be measured by the flow cytometric profile using the same interpretation tools as in cell cycle analysis.
The cell permeant DNA-specific dye DRAQ5™ has no spectral overlap with R-PE and FITC making it easy to perform multiparametric cell subset analysis. It can be used with live or fixed cells.
To optimize 3- and 4-colour analysis of lymphoma cell preparations on a single laser (488 nm) equiped benchtop flow cytometer, (Plander et al. 2003 ) demonstrated that with DRAQ5™ cell cycle and ploidy studies could be performed with FITC, PE & ECD conjugated antibodies. Optimal results, comparable to Propidium Iodide, were obtained with fixed samples. However, improved c.v.'s for live cells were obtained by incubating with DRAQ5™ for 30 minutes.
EXAMPLE:
Ploidy analysis in small cell lung carcinoma: CD45- cells (R1) compared to CD45+ / SSClow (R2, T cells). Invasive CD45 negative cells found to be hyperploid. Data courtesy of Stephen Couzens, Haematology, University Hospital of Wales, Cardiff.
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