Simple Flow-Based Total Nucleated Cell Differential

A new method published in Cytometry part B uses DRAQ5 to gate all nucleated cells, FlowCount beads, a panel of 6 antibodies in 4 clusters and the 5 fluorescence channels plus forward and side scatter on the Beckman Coulter FC500 flow cytometer to get a comprehensive analysis of both nucleated cells and platelets in blood or bone marrow samples. Ten separate classes of nucleated cells could be identified.

The one-tube, lyse-no wash method gave correlation to both cell counters and microscopy and offers a real alternative to the laborious manual scoring of blood smears from samples flagged by the cell counter.

Using the DNA binding properties of DRAQ5 cell cycle information was also available whilst gating on the DRAQ5-/CD36+ events permitted enumeration of platelets. This method could offer an effective replacement for the subjective, labour-intensive and statistically-limited analysis by manual microscopy.