Comments on: I’ve worked out a method for doing immunohistochemistry (IHC) on formalin-fixed paraffin embedded (FFPE) sections of bone. http://synthesis.williamgunn.org/2007/08/31/ive-worked-out-a-method-for-doing-immunohistochemistry-ihc-on-formalin-fixed-paraffin-embedded-ffpe-sections-of-bone/ A synthesis of ideas about open science and social technology. Wed, 18 Jan 2012 20:57:43 +0000 hourly 1 http://wordpress.org/?v=3.3.1 By: Mr. Gunn http://synthesis.williamgunn.org/2007/08/31/ive-worked-out-a-method-for-doing-immunohistochemistry-ihc-on-formalin-fixed-paraffin-embedded-ffpe-sections-of-bone/comment-page-1/#comment-15771 Mr. Gunn Mon, 23 May 2011 00:36:05 +0000 http://synthesis.williamgunn.org/2007/08/31/ive-worked-out-a-method-for-doing-immunohistochemistry-ihc-on-formalin-fixed-paraffin-embedded-ffpe-sections-of-bone/#comment-15771 I'm delighted to have the question, John! My procedure was really simple. I did a xylene dip, followed by 100%, 95%, 70% EtOH and then DH2O. I didn't do antigen retrieval, as that destroyed the morphology of my thin sections and tended to kill the TRAP activity I was staining for as well. My goal was to get a osteoblast/osteoclast ratio, and I ended up consulting with a pathologist for that as the method above clearly demonstrated osteoclast activity, but didn't lend itself to quantitative analysis. I detailed the work in my dissertation <a href="http://williamgunn.org/Dissertation%20William%20Gunn.pdf" rel="nofollow">here</a>. If you'd tell me a little about what you're trying to achieve, perhaps I could make some further suggestions. I’m delighted to have the question, John! My procedure was really simple. I did a xylene dip, followed by 100%, 95%, 70% EtOH and then DH2O. I didn’t do antigen retrieval, as that destroyed the morphology of my thin sections and tended to kill the TRAP activity I was staining for as well.

My goal was to get a osteoblast/osteoclast ratio, and I ended up consulting with a pathologist for that as the method above clearly demonstrated osteoclast activity, but didn’t lend itself to quantitative analysis. I detailed the work in my dissertation here. If you’d tell me a little about what you’re trying to achieve, perhaps I could make some further suggestions.

]]> By: John http://synthesis.williamgunn.org/2007/08/31/ive-worked-out-a-method-for-doing-immunohistochemistry-ihc-on-formalin-fixed-paraffin-embedded-ffpe-sections-of-bone/comment-page-1/#comment-15770 John Sat, 21 May 2011 20:37:13 +0000 http://synthesis.williamgunn.org/2007/08/31/ive-worked-out-a-method-for-doing-immunohistochemistry-ihc-on-formalin-fixed-paraffin-embedded-ffpe-sections-of-bone/#comment-15770 Hi, I'm currently trying to do the same type of staining on decalicified mouse bone embedded in paraffin (Fixed in Formalin). I was wondering if you could communicate here your optimal Antigen retrieval/deparaffin protocol (or at least the major steps) for immunofluorescence staining (NON ENZYMATIC) Many thanks in advance. Best John Hi, I’m currently trying to do the same type of staining on decalicified mouse bone embedded in paraffin (Fixed in Formalin).

I was wondering if you could communicate here your optimal Antigen retrieval/deparaffin protocol (or at least the major steps) for immunofluorescence staining (NON ENZYMATIC)

Many thanks in advance.

Best

John

]]> By: Mr. Gunn http://synthesis.williamgunn.org/2007/08/31/ive-worked-out-a-method-for-doing-immunohistochemistry-ihc-on-formalin-fixed-paraffin-embedded-ffpe-sections-of-bone/comment-page-1/#comment-445 Mr. Gunn Thu, 06 Dec 2007 17:24:46 +0000 http://synthesis.williamgunn.org/2007/08/31/ive-worked-out-a-method-for-doing-immunohistochemistry-ihc-on-formalin-fixed-paraffin-embedded-ffpe-sections-of-bone/#comment-445 Yes, I have some of the pictures at <a href="http://www.ihcworld.com/smf/index.php/topic,1973.msg5381.html#msg5381" rel="nofollow">ihcworld</a>. I never tried it on calcified frozen sections, but I don't see why it wouldn't work. Yes, I have some of the pictures at ihcworld. I never tried it on calcified frozen sections, but I don’t see why it wouldn’t work.

]]> By: Orit Kollet http://synthesis.williamgunn.org/2007/08/31/ive-worked-out-a-method-for-doing-immunohistochemistry-ihc-on-formalin-fixed-paraffin-embedded-ffpe-sections-of-bone/comment-page-1/#comment-444 Orit Kollet Thu, 06 Dec 2007 16:21:13 +0000 http://synthesis.williamgunn.org/2007/08/31/ive-worked-out-a-method-for-doing-immunohistochemistry-ihc-on-formalin-fixed-paraffin-embedded-ffpe-sections-of-bone/#comment-444 Dear Mr. Gunn, I found your Post very interesting and would like to know if you have already uploaded any pictures of your staining. Did you ever tried to use your method on frozen calcified bone sections? Best, Orit Dear Mr. Gunn,

I found your Post very interesting and would like to know if you have already uploaded any pictures of your staining. Did you ever tried to use your method on frozen calcified bone sections?

Best, Orit

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