RE: Use of 7-AAD on a FACS Array, with fixation??

Hi Patricia,
 
You could try the LIVE/DEAD Fixable Red Dead Cell Kit from Molecular
Probes. The excitation / emission maxima of the dyes are about 595/615.
Although we have not tested the dye with green excitation, there is a
sufficient amount of excitation in that region based on the spectrum
that it should distinguish dead cells.
 
Regards,

Bill

 

William Godfrey, Ph.D.

Flow Cytometry Research Area Manager

Invitrogen Corp.

29851 Willow Creek Road

Eugene, OR 97405

office: 541-335-0141

cell: 541-729-5316

 

________________________________

From: Guy Hermans [mailto:Guy.Hermans@ablynx.com] 
Sent: Thursday, May 26, 2005 12:27 AM
To: cyto-inbox
Subject: RE: Use of 7-AAD on a FACS Array, with fixation??



Hi Patricia,

 

The Array resuspended cells prior to sample uptake by "pipetting" up and
down (see user prefs), then "pipette up" a given volume and injects this
into the sample pathway, which is a closed cuvette-straight-to-waste
type of deal. I guess there are no aerosols involved in the same sense
as in sorters/jet-in-air devices, maybe a tiny bit if you're trying to
"pipette up" volumes larger than the wells actually contains.

 

I mostly use PI or TOPRO3 on the Array, have done 7-AAD sometimes -
never tried combining any of those with fixation, as my understanding is
none of these can be used on fixed cells: everything will probably light
up.

 

Guy

 

 

Next generation therapeutic antibodies <http://www.ablynx.com> 

 

Guy Hermans, PhD
Project Leader 

Ablynx NV
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Belgium 

guy.hermans@ablynx.com <mailto:guy.hermans@ablynx.com>  

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-----Original Message-----
From: Patricia Echeagaray [mailto:pechado@hotmail.com] 
Sent: Wednesday, May 25, 2005 6:22 PM
To: cyto-inbox
Subject: Use of 7-AAD on a FACS Array, with fixation??

 


All,

Doing a new procedure and a question came up.....

Is it possible to use 7-AAD on a Facs Array, using infected cells WITH
fixation? 
OR must they be analyzed without the benefit of being fixed? 
Does the FacsArray aerosolize (sp?) the sample?
Does 7-AAD tolerate any fixation ? 

TIA!!

Patti in damp. rainy MD 
(but the flowers (as in plants) are happy, if the flowers (as in
cytometrists) are not!)