[Cytometry] Analysis of prawn cells

John Wilson j.wilson3 at uq.edu.au
Mon Feb 9 01:35:52 EST 2009


Hi Everyone,
 
I have a customer who is interested in analysing prawn blood or haemolymph cells by flow cytometry. This is done by plotting lin FSC vs log SSC. Reportedly 3 populations should be clearly visible. The problem we have is there is very little reproducibility between animals (positional shifts and obscure populations). If anyone has a protocol specific for prawn blood cells or has has experience to share that would be appreciated.
 
The current protocol is a harvest via heart puncture. Cells are then suspended in Alsevers solution (vertebrate anti-coagulant pH 4.6) and then run on an LSR II. There is a delay of approximately 1-2 hours between anti-coag and analysis where cells are kept on ice.
 
Alsevers solution has been tried at pH7.0, but this gives very poor reproducibility and also poor separation on FSC vs SSC. This also occurred when EDTA at pH 7.0 and pH 4.6 was tried.
 
Thanks,
 
John Wilson
Flow Cytometry Laboratory
Queensland Brain Institute
The University of Queensland
<mailto:j.wilson3 at uq.edu.au>  


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