Ian_DIMMICK at europe.bd.com
Wed Jun 2 17:23:17 EST 2004
I have worked with platelets and find the following guidelines useful,
1 Establish the correct gate by running a normal sample of whole blood
, usually on log FS/SS, it is useful to stain the platelets with a
glycoprotein marker (CD61) to establish that the scatter gate does contain
a reasonably pure population of platelets
2 Beware of thresholds, by using log scatter parameters thresholds cam
come in very high on the axis even with a low threshold value dependant
upon which instrument you use
3 The concentration of platelets can be high , also the surface is very
rich in glycoprotein and has coincidentally a high non specific binding, be
aware that you have to avoid antigen excess
4 The reporting of intensity of CD41b can be calibrated as can most
other antigens with commercially available standards
5 Platelet aggregates can only be minimized , usually I use EDTA in PBS
the same concentration as you would use to anticoagulate whole venous blood
, in fact the method I use is to add PBS to an EDTA blood bottle to the
recommended volume for blood and use that as EDTA and PBS, very simple
6 Try talking to them nicely and always tell them the truth
"Elham Ashouri" <ashourie at pearl. To: Cytometry Mailing List
<cytometry at flowcyt.cyto.purdue.edu>
sums.ac.ir> cc: (bcc: Ian DIMMICK/Europe) Subject: platelet 02/06/2004 08:12 Please respond to ashourie
I try to work with platelet . I am going to run platelet sample.
Some questions about it:
1.if I have low platelet number then can I analysis it as usual?
how can I find correct gate?
2.how can I report the intensity of CD41b(percent, MFI)?
3.if the sample continuing jaint or aggregate platelet how can I
Looking forward to hearing from you soon
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