[Histonet] IHC without wash buffer

[Goins, Tresa]

In a continuing effort to limit the volume of reagent used in each step of a manual IHC, I have tried TBS and TBST on slides with barriers (expensive) and without barriers.
The results were not stellar with the barrier slides - the reagent still escapes.  We dewax with hot detergent (may be a contributing factor) and the Tween 20 in the TBST definitely alters the hydrophobicity of the barrier, an effect that is not reversed with a water wash.

Consequently, I have resorted to omitting the buffer wash steps and using distilled water only.  The slide surface remains "water repellent" and the added IHC reagents form a pool over the tissue sections.
The detection method is a polymer-HRP and there is no increase in background staining in the tissue or on the slide surface.

I am assuming that the IHC reagents are prepared in the "optimum" suspension liquid and a buffer is not required.  So, I am interested in hearing from anyone why this is a bad idea.

Thanks,

Tresa






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