[Histonet] RE: Eosin too pink

Mark Adam Tarango marktarango <@t> earthlink.net
Sat Jan 14 20:03:56 CST 2006

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

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