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Author Topic: Help with Eosin staining  (Read 2531 times)

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Offline waheedah

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Help with Eosin staining
« on: June 17, 2010, 11:41:10 AM »
Dear All,

This is my first time here and I am new to immunostaining techniques as well. I am sorry if this may be a repeat question...

I am trying to perform a H&E staining on lung tissues to visualize its morphology for laser tissue micro dissection.

I am supposed to stain the 10mm tissue section and dehydrate it in serial ethanol within 3-5 minutes.

The hemotoxylin is fine but the aqueous eosin that I use (after 30s staining) de-stains completely in 70& ethanol. Is there a way to 'fix' the eosin staining in ethanolic solutions?

I would be grateful for your help.

Thank you.

Help with Eosin staining
« on: June 17, 2010, 11:41:10 AM »

Offline hardrob

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Re: Help with Eosin staining
« Reply #1 on: June 18, 2010, 04:21:58 AM »
in our lab, we use a Eosine- solution with 0.5% for 1 minute. after that the slides go directly in acetic acid (1%) for 5 seconds following ethanol 70%-96%- 100%-100%-Xylene- Xylene and it works fine. just stay only for a second or so in ethanol 70% and go on to 96, becaus the water in the 70 dilutes the eosine.

Offline richard03

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Re: Help with Eosin staining
« Reply #2 on: June 18, 2010, 12:53:43 PM »
Skip 70% ethanol and go straight to 95% ethanol for only 10 seconds or 10 dips but no more than 30 seconds. Then transfer to 100% ethanol and you will be safe.

Offline excalibur

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Re: Help with Eosin staining
« Reply #3 on: June 20, 2010, 11:39:23 PM »
I use an alcoholic eosin and go straight to 100% EtOH after staining.
Paula Keene Pierce, HTL(ASCP)HT
Excalibur Pathology, Inc.
5830 N Blue Lake Dr.
Norman, OK 73069
405-759-3959
www.excaliburpathology.com

Re: Help with Eosin staining
« Reply #3 on: June 20, 2010, 11:39:23 PM »