[Cytometry] Whole Blood Samples VS Cyan ADP

Tomas Adejumo t.adejumo at ucl.ac.uk
Mon Feb 9 11:30:18 EST 2009


Dear All,

We currently use a Cyan ADP from Beckman Coulter and one of my
colleagues is analysing blood samples and prepares the cells using a
lysing kit and protocol, e.g.

Reagent 'A'
50ml H20 
60ul Formic acid.

Reagent 'B'
6g/l Na2Co3
14.5g/l NaCl
31.3g/l Na2SO4

Reagent 'C'
10% Formaldehyde (from paraformaldehye solid) in Reagent 'B'

Use all reagents at 0'C

Procedure.

i)	Add 5ul ab/50ul whole blood

ii)	Incubate on ice 20mins

iii)	Add 300ul 'A' -Vortex/10 seconds.

iv)	Add 132.5ul 'B' -Vortex/10 seconds.

v)	Add 50ul 'C' -Vortex/10 seconds.

> Keep samples in the 'fridge' or on ice -should be stable for around a
week
> Run on the flow.

However, on running the samples, some of them appear to block the SIP of
the Cyan. We have checked for pressure problems etc but there are none.
And when running some of her samples, she doesn't experience this. The
user doesn't wish to filter the samples but does use EDTA and heparin.

When running the same sample on the FACS Calibur, she doesn't notice
this problem.

Can you suggest why that is?

Cheers, Tomas

Tomas Adejumo MSc
Applications Specialist - Flow Cytometry and Confocal Microscopy
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