Laser Scanning Slide & Microplate Cytometry whole blood differential
ideal "trigger" for cell analysis....
In a 10 µl whole blood method DRAQ5™ makes the essential clear distinction between leukocytes and erythrocytes.
The combination of a Neubauer counting chamber to give a known volume and the Laser Scanning Cytometer to give the cell immunophenotype and scatter information allow analysis of a 10µl whole blood sample.
DRAQ5™ was used as a "trigger" for the leukocyte analysis. It exclusively stains nucleated cells (live or dead) and its emission is spectrally separated from that of FITC and PE fluorochromes.
In short, 10 µl of EDTA blood was diluted with 10 µl of CD45-FITC and CD14-PE antibodies in PBS, and 10 µl of a 500 µM working solution of DRAQ5™.
It was also possible using the method to get an erythrocyte count. The sample was diluted a further 150-fold, sample placed in the counting chamber and erythrocytes detected using a forward scatter gate.
The method provides a means of getting important haematological information from small sample volumes, essential in situations where sampling is difficult or traumatic (e.g. neonates).
Reference
Mittag A, et al. Small and cheap: accurate differential blood count with minimal sample volume by laser scanning cytometry (LSC). Proc. SPIE 2005; 5692:79-90.
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