RE: [Histonet] IHC unclear at 40x

From:koellingr@comcast.net



Bernard,
All the suggestions I saw, oil, dehydration, dry, Koehler illumination are great.  Is another possibility that you have a correction collar (like for coverslip thickness) on the barrel of your 40x?  Some don't but many do.  It is easy to be clear at 40x and purposely move the correction collar to become blurry.  If you don't know its there, it just might be where it has been placed and has been all along and needs to be adjusted by simply slightly revolving into focus

Ray Koelling
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 
From: "Jo Dee Fish"  

> Bernard, 
> Don't forget to do Koehler illumination procedure to your microscope as 
> well. If you need instructions, try here: 
> http://www.rawlight.com/Koehler_illumination.pdf 
> Take care, 
> Jo Dee 
> 
> 
> ~~Jo Dee Fish~~ 
> Research Technologist III 
> Gladstone Institute of Cardiovascular Disease 
> 
> Telephone: (415) 734-2567 
> Fax: (415) 355-0824 
> E-mail: jfish@gladstone.ucsf.edu 
> 
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> The J. David Gladstone Institutes 
> 1650 Owens Street 
> San Francisco, CA 94158 
> 
> -----Original Message----- 
> From: histonet-bounces@lists.utsouthwestern.edu 
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa 
> Sent: Friday, February 08, 2008 12:46 PM 
> To: Bernard Martin; histonet@lists.utsouthwestern.edu 
> Subject: Re: [Histonet] IHC unclear at 40x 
> 
> Make sure that when you finish and dehydrate to coverslip, that dehydration 
> is complete before clearing the section with xylene. 
> RenͬJ. 
> 
> Bernard Martin wrote: 
> My IHC looks great at 5x, 10x, and 20x, but when I go up to 40x, it is a 
> bit blurry. The objective is clean, as is the slide itself. I deparaffinate 
> in clean xylene. Anyone run into this problem, and if so, how did you 
> resolve it? Thanks! 
> 
> 
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