Re: platelet

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


Ian


		      "Elham Ashouri"							  		      <ashourie@pearl.	       To:	Cytometry Mailing List
<cytometry@flowcyt.cyto.purdue.edu>			
		      sums.ac.ir>	       cc:	(bcc: Ian DIMMICK/Europe)	  					       Subject: platelet			  		      02/06/2004 08:12							  		      Please respond							  		      to ashourie							  




Dear All,

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
analysis that?

Looking forward to hearing from you soon

Regards,

Elham