[Histonet] Re: Post fixing IHC slides

[Obregon, Cecilia]

We bake our slide in the oven for 30 minutes at 65 degrees. 

Then placed them in a staining bucket 'covered' at room temperature for 30 minutes. This small bucket contains a piece of gauze soaked in formalin at the bottom. 

We do not 'immerse' the slides in formalin, we just sort of use the contained fumes to post fix the slides. No rinsing after this step, just load them in the Benchmark Ultra, and coverslip as usual. (We do manually rinse all of our slides in Di soapy water before coverslipping..whether they have been post fixed or not to remove excess LCS)

Hope this helps,

I can be reached at 954-265-5317 if you have any further questions.

Thanks-Cecilia

________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of abtdhu <@t> gmail.com [abtdhu <@t> gmail.com]
Sent: Monday, March 30, 2015 8:30 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Post fixing IHC slides

Hi Cecilia,
Glad to know post fixation will reduce section falling off from slide. We are bone research lab and always have problem of paraffin bone section falling off after antigen retrieval. There are many ways we have tried, but not improve too much. I know bone section will be easy to fall if it was incompletely fixed. I want to try your post fixation to secure the bone section on the slide. Would you tell me the exact way you are doing? You bake the slides in oven and deparaffin first? Then post fix the slides 30 minutes? How do you do vapor fixation? Then go on for IHC after rinse when you done post fixation?
Your answer is very much appreciated.
Dorothy

> 1. RE: Post fixing IHC slides in formalin (Obregon, Cecilia)
>
> From: "Obregon, Cecilia" <CObregon <@t> mhs.net>
> Subject: RE: [Histonet] Post fixing IHC slides in formalin
>
>
> Our facility implemented the post fixing of IHC slides years ago only for tissues like breast, cell blocks, or anything that tends to fall off. We baked the slides in the oven for 30 minutes, and then post fixed them for an additional 30 min in formalin 'fumes'. It works great on breast resections specimens, and it doesn't interfere with staining.
> Thank you,
>
> Cecilia M. Obregon
> Memorial Regional Hospital
> 3501 Johsnon Street
> Hollywood, FL 33021
>
> ________________________________________
> From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] on behalf of Barry Rittman [barryrittman <@t> gmail.com]
> Sent: Saturday, March 21, 2015 2:20 PM
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] Post fixing IHC slides in formalin
>
> There are always discussions about fixation but I have never seen comments
> about using vapor fixation for post fixing or for fixing fresh frozen
> specimens.
> Vapor fixation is simple to use and does not require any solvent as it uses
> the tissue fluids in the case of fresh frozen sections or the solution
> remaining after IHC.
> Especially useful for formaldehyde, alcohol,  acetone, osmium tetroxide etc.
> Barry
>
>> On Sat, Mar 21, 2015 at 12:14 PM, Ann Specian <thisisann <@t> aol.com> wrote:
>>
>> Does anyone post fix their IHC slides in formalin in an effort to try to
>> reduce tissue loss?  If so, does anyone have a protocol for this that they
>> have used and have seen good results?
>>
>>
>> If you have any other suggestions which can help to reduce tissue loss
>> during IHC staining, I would love to hear from you.
>> Thanks,
>> Ann
>> _______________________________________________
>>

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