[Histonet] RE: Eosin too pink

[Mark Adam Tarango]

So is he complaining that the eosin is too strong or just too pink?  It seems to me like the more water you have in the alcohol after Eosin or if you use a water rinse after eosin it gets pinker....it's more orange when you have a higher percentage of alcohol in the alcohol after eosin(just seems that way to me anyway).  If it's just too strong, dilute it down with alcohol.  I hardly get any specimens, so I don't like changing the Eosin often, i just keep adding alcohol to it when it gets low and crusted around the edges..... still seems to work fine....

Mark T.

> Sent: Friday, January 13, 2006 10:42 AM
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Eosin too pink
> 
> Hi List,
>  We are currently doing our H & E stain by hand and using the
> Richard-Allan 7211 Hematoxylin and their Eosin Y.  Our pathologist is
> still complaining of slides being TOO PINK.  I have cut the eosin step
> down to just 5 dips from 15 dips, and still too pink.  The Eosin is very
> rich. 
> 
> This is our protocol:
> Hydrate to water like normal (thru Clear Rite 3, 100% and 95% Flex) 
> Hematoxylin- 2 mins
> Rinsing in water until the water runs clear
> Clarifier II- 30 dips
> running water -30 dips
> Richard Allan Bluing - 30 dips
> running water - 30 dips  
> 95% Flex - 10 dips
> Eosin-Y- 5 quick dips
> Thru 95%, 100% etc to xylene
> 
> Any suggestions on a possible method change or a more mild Eosin.
> Complaint seems to solely "TOO PINK".  I have done some experimenting w/
> cutting the eosin w/ 80% Flex- and it does calm it down a bit but, the
> Hemo looks more purply then blue, almost periwinkle so not sure if that
> will be better or not- we still need to have him review the test slides.
> Any suggestions ?
> 
> Helayne Parker
> Histology Section Head
> Skaggs Community Health Center
> Branson, Missouri