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Hi Fredrik,

I also get this kind of staining in mouse, and dog heart ventricle cells. In
the Fitc range, I think that can be autofluorescence.  there was a subject
on this list 1 or 2 years ago.  I will take a look in the archives...

Also, i have seen these kind of pattern  with other wavelenths (triTC, Cy5)
and the negative controls were looks good.  For these experiments, I was
blocking the isolated cells with 10%serum.  I have reduced my blocking
buffer concentration (to +/- 2% serum) and the recurrency of this staining
had dissapeared.  does too much blocking can cause non specific stainng?
This is an option that I leave open...

Best regards

Louis

Louis R. Villeneuve
Assistant de recherche -microscope  confocal
Institut de Cardiologie de Montréal- Centre de Recherche
5000 est Bélanger, Montréal
Québec , Canada
H1T 1C8

514-376-3330 ext3511
514-376-1355 (Fax)
----- Original Message ----- 
From: "Fredrik Swift" <[log in to unmask]>
To: <[log in to unmask]>
Sent: Wednesday, January 21, 2004 5:15 AM
Subject: Cardiomyocytes


> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hello,
>
> I'm labelling rat cardiomyocytes with different antibodies. Sometimes I
get
> what I think is unspecific staining. The staining consists of longitudinal
> "strips" with low fluorescence intensity (Alexa-488 labelled 2nd ab's).
This
> is typical if I use very low concentrations of primary ab's, and I get
> similar results in negative control exp. with no primary ab. Apparently,
the
> 2ary ab is the cause of this, but I cannot explain what it stains. Is it
> myosin/actin that I see? Have anyone had the same results previously? Any
> help would be appreciated! I can send jpg.-files of the images if the
> description wasn't good enough.
>
> Regards,
> Fredrik