Fixation --- Labeling ONLY surface molecules

JOHN BOUCHER jbouch at verizon.net
Fri Feb 15 20:20:22 EST 2008


Elise, you took the words right out of my mouth.  I ,too, need to  
find a technique for overcoming the membrane leakiness caused by  
fixing RBCs (glutaraldehyde or formaldehyde) in order to label only  
surface molecules.

Improving the fixation technique to prevent/inhibit membrane pores,  
OR, "building" a fluorescent label larger than the pores so that they  
won't penetrate the membrane is a long sought goal.

Please, please, if someone answers you with a miracle answer, I'd  
appreciate knowing.

A couple of months ago, Vicki Singer (Invitrogen)  
(<vicki.singer at invitrogen.com>) notified me that Invitrogen might be  
interested in looking into a solution for "getting around" the	
leakiness of cell membranes caused by fixatives.  If Vicki is  
listening, I hope she'll let us know the status of Invitrogen's  
possible solution.

------------------------------------------
John H. Boucher, DVM, PhD
Rheotech Labs, Inc.
Comparative Studies in Blood Rheology
2106 Salisbury Road
Silver Spring, MD 20910, U.S.A.
Phone:	301-585-1885
Email:	<jbouch at verizon.net>
========================================================================
On Feb 14, 2008, at 5:01 PM, Palmer, Elise wrote:
> Hi,
>
> I was wondering if anyone has experience assessing apoptosis with  
> annexin V and 7AAD, and fixation?
>
> I need to run some HIV patient samples so they have to be fixed. I  
> will be looking at lymphocytes. I have had no success finding  
> anything in the literature in which both annexin V and 7AAD were used.
>
> I would really appreciate any ideas or help you can give me!
>
> Elise (University of Rochester Medical Center)
>
>

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