[Histonet] replacing tween in the secondary hyb.solution

[Geert van Haalem]

Tween indeed facilitates the dispersion. I was testing the Ventana autostainer and I got some inconsistent
staining with patchiness of certain features instead of a homogeneous staining.  I checked the presence and
absence of tween to check if it somehow interferes with the liquid coverslip of Ventana. I just found out that
also with a Z-stack that a significant depth variation is created somewhere during the run. It still just doesn't
compete with the manual staining.
Presumably the reason I could not find by eye a difference between with or without tween is that only
100 microliter is dispensed on the slide with on top the liquid coverslip and thus forcing the dispersion over
the section.



From: Rene J Buesa [mailto:rjbuesa <@t> yahoo.com]
Sent: Monday, March 05, 2012 4:46 PM
To: histonet <@t> lists.utsouthwestern.edu; Geert van Haalem
Subject: Re: [Histonet] replacing tween in the secondary hyb.solution

Not a good idea. The effect of Twenn is to facilitate the dispersion over the section of the reagent. If you do not use it the reagents may not reach all the surface and you may end with an uneven reaction or your results could be inconsistent. A small savings by not using Tween may mean that you could have to repeat the test and spend much more expensive reagents. Again, not a good idea.
René J.

--- On Mon, 3/5/12, Geert van Haalem <g.vanhaalem <@t> prosensa.nl<mailto:g.vanhaalem <@t> prosensa.nl>> wrote:

From: Geert van Haalem <g.vanhaalem <@t> prosensa.nl<mailto:g.vanhaalem <@t> prosensa.nl>>
Subject: [Histonet] replacing tween in the secondary hyb.solution
To: "histonet <@t> lists.utsouthwestern.edu<mailto:histonet <@t> lists.utsouthwestern.edu>" <histonet <@t> lists.utsouthwestern.edu<mailto:histonet <@t> lists.utsouthwestern.edu>>
Date: Monday, March 5, 2012, 10:25 AM
Hi,

I currently did an experiment in which I left out Tween in de hybridisation solution of
the secondary antibodies. I only made a solution of two Alexa in PBS. When imaging
Bye eye I did not notice any decrease in signal compared to a hyb.solution with Tween.
My intention is to replace tween with FBS, but can anybody tell me pro and cons for
this idea.

With regards,

Geert
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu<http://us.mc1621.mail.yahoo.com/mc/compose?to=Histonet@lists.utsouthwestern.edu>
http://lists.utsouthwestern.edu/mailman/listinfo/histonet