Re: DRAQ5: may be an alternative for Hoechst 33342 if I have only 488nm argon laser?

From: Pál Jáksó (Pal.Jakso@aok.pte.hu)
Date: Mon May 06 2002 - 02:26:51 EST


Hi Bernie,

As you could see it was a question from me and I'm new too using DRAQ5.
My first results show that DRAQ5 can not be used for aneuploidy
measurements combined with surface immunostaning in heterogen samples
like blood and bone marrow because of the different staning intensity of
the different type of cells (e.g. granulocytes show DNA index: 1,1
compared to lymphocytes). However in my opinion it could be useful in
homogen samples like cell lines. The CV% was not bad (about 3-5%) but PI
shows better resolution.
In the end DRAQ5 proved very useful when I used it as tresholding
parameter for exluding unnucleated cells and debris. For example I got
nice results with absolute CD34+ cell counting combining DRAQ5/CD45
FITC/CD34PE/BD's TruCount tubes with lyes-no-wash method. It seems to be
an economic alternative of BD's ProCount kit.
Otherwise what do you want to use DRAQ5?

Bests,

Pal

On Saturday, May 4, 2002, at 12:45 , Fallon, Bernadette wrote:

> Hi,
> I am just wondering if you had much sucess with the Draq 5? I am
> starting
> out using it and results so far are awful....
> Bernie Fallon
>
> -----Original Message-----
> From: JˇksŰ Pˇl [mailto:Pal.Jakso@aok.pte.hu]
> Sent: Wednesday, March 06, 2002 3:43 AM
> To: Cytometry Mailing List
> Subject: DRAQ5: may be an alternative for Hoechst 33342 if I have only
> 488nm argon laser?
>
>
>
> Dear flowers,
>
> I read a post from Derek Davis in which DRAQ5  a far red emitting DNA
> stain from Biostatus was recommended for DNA staining for flow cytometry
> without permeabilization of living cells. I would like to ask about your
> experiences with this new DNA dye.
> I intend to measure surface immunostainings in conjuctions with DNA
> content using 488 nm argon laser. At the Biostaus website is stated that
> using together with FITC compensation is not necessary and DRAQ5 can be
> also used with PE.
>
> Thanks in advance,
>
> Pal Jakso
>
> University of Pecs
> Faculty of Medicine
> Dept. of Pathology
> 7643, Hungary
> 12. Szigeti str.
>



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