[Histonet] Strange circles in IHC slides

[V. Neubert]

>
> It looks like bubble trapped between the slide and coverplate.  Are you using
> sufficient surfactant?  That can help, but any of the "cap gap" methodologies
> lend themselves to this problem.
>
> Rinse your slides very thoroughly in buffer with plenty of surfactant before
> loading them on the instrument.
Actually, I don't use any specific surfactant, as long as it is not 
included in TBS buffer ;)
What do you recommend? Maybe just rinsing in buffer would do? I'll give 
it a try tomorrow, I think.

> The bubbles to which I refer to are not coverslipping bubbles,
> rather bubbles that were present on the sections during staining that
> keep the antibody(ies) and/or the chromogen from making contact with the
> tissues. Still, I would think if having that many slides affected, you
> probably would have noticed an inordinate amount of bubbles present
> between the coverplates and the slides at the time of staining!?! 
How can they develop? I did not see any bubbles, because I don't change 
them between the steps.

Thanks for your replies :)