[Histonet] Eosin too pink

Parker, Helayne HParker <@t> Skaggs.Net
Fri Jan 13 10:42:11 CST 2006

 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

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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