[Histonet] Need Suggestions: IHC Slides and Floating Sections + 4% PFA Fixation

[Nasir Abbas via Histonet]

Hi All
1) How long vibratom liver sections when cut in ice cold PBS and then stored at 
4C in PBS could be used? 2) How long paraffin embedded liver tissues after 
dewaxing and antigen retrieval stored in PBS at 4C could be used?
3) Whether liver tissues fixed in 4% PFA for more than a month would have any 
affect on antigen integrity?when antigen of interests (CD68, F4/80, Ly6g, CD4 
and CD8) are low in abundance (different time points from 15min to 96hrs of 
injury)? Has any body encountered such problem before? If such was the case 
with you, how would have you tackle it?
and finally,3) Any body could suggest liver pathology atlas with normal to 
special stain please?
For your opinions and suggestions, thanks in advance.
Sincerely Nasir Abbas
---PhD FellowGuangzhou Institutes of Biomedicine and HealthChinese Academy of 
Sciences190 Kai Yuan Avenue Science Park Guangzhou, 510530, Guangdong 
P.R.ChinaCell: +86-1312 8296 614
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[Histonet] Suggestions: Liver Pathology Book/Atlas

[Nasir Abbas via Histonet]

Hi Histo-FamilyI wonder if some one could suggest an updated and best Liver 
Pathology Book or Atlas with slight explanation, which covers all pathology 
assays. Or if some one already have for sale (used one), that will be 
preferred. Any suggestions please?
Sincerely 
Nasir Abbas
---PhD FellowGuangzhou Institutes of Biomedicine and HealthChinese Academy of 
Sciences190 Kai Yuan Avenue Science Park Guangzhou, 510530, Guangdong 
P.R.ChinaCell: +86-1312 8296 614
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[Histonet] Suggestion for Dual staining Kit/reagent

[Nasir Abbas via Histonet]

Hi All
Can some one suggest dual staining reagents for IHC-P (mouse liver tissue) for 
my primary antibodies: ab183685, GTX53126 and ab25377.I was suggested vectors 
labs products (following table) but I am not clear about Vector Blue reagent 
SK-5300 that whether it can be used in IHC-P, because it features shows it is 
fluorescent. Moreover it is sensitive to xylene and Histoclear may be used as a 
clearing reagent in this case, which makes me even more worried. Any suggestion 
and tips for my dual staining?Table:
| Pri-Ab | Vector Lab Products- Suggested |
| ab183685 | MP-7401-15 |
| GTX53126 | SK-4105 |
| ab25377 | MP-5404-15 |
|   | SK-5300 |


 
Sincerely Nasir Abbas
---PhD FellowGuangzhou Institutes of Biomedicine and HealthChinese Academy of 
Sciences190 Kai Yuan Avenue Science Park Guangzhou, 510530, Guangdong 
P.R.ChinaCell: +86-1312 8296 614
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Re: [Histonet] Histonet Digest, Vol 154, Issue 22

[Nasir Abbas via Histonet]

Dear Histoneters!Good DayMay I request you guys to share protocol of frozen 
tissues (mouse liver) for immunoflorescence assay. Lots of protocol are 
available online but i will highly appreciate the one that could lead me from 
tissue preparation towards staining procedure as I am really blank about frozen 
tissues and slides dealing. So I will be on standby for your response. Thanks 
in advance.
Sincerely Nasir Abbas
---PhD FellowGuangzhou Institutes of Biomedicine and HealthChinese Academy of 
Sciences190 Kai Yuan Avenue Science Park Guangzhou, 510530, Guangdong 
P.R.ChinaCell: +86-1312 8296 614 

On Wednesday, 28 September 2016, 1:12, 
"histonet-requ...@lists.utsouthwestern.edu" 
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Today's Topics:

  1. GMS (Martin, Gary)
  2. Re: Unprocessing tissue (Caroline Miller)
  3. Water for H Stainers? (P Sicurello)
  4. Re: Water for H Stainers? (Elizabeth Chlipala)


--

Message: 1
Date: Mon, 26 Sep 2016 16:30:49 -0700
From: "Martin, Gary" 
To: 
Subject: [Histonet] GMS
Message-ID:
    <6ED9D4252F278841A0593D3D788AF24C366E5E51@mailsvr.MARSHMED.local>
Content-Type: text/plain;    charset="us-ascii"

I am having difficulties with a stain I've done many times ...GMS. The
light green is only staining the outer edges of the specimen. Any Ideas?


Thanks 

Gary



--

Message: 2
Date: Mon, 26 Sep 2016 16:49:14 -0700
From: Caroline Miller 
To: "Armstrong, Karoleigh T" 
Cc: "histonet@lists.utsouthwestern.edu"
    
Subject: Re: [Histonet] Unprocessing tissue
Message-ID:
    

[Histonet] Markers for Kupffer cell: Secondary Conjugate Systems

[Nasir Abbas via Histonet]

Dear All,
I need your guidance in the following issues,
Issue No:1I am going to identify two markers on a single cell (macrophage) as 
well as on two different cells (CD4 and CD8) in a single tissue (costaining). 
What could be the best secondary conjugate systems for detection in this case 
(I only have some experience with HRP conjugate). A protocol will be 
appreciated.
Issue No.2what are the specific markers of Kupffer cells?
I am working with CD11b and F4/80 for said cells but it seems (found in 
articles) that these two markers are also expressed by other leukocytes as 
well, but my job require a very specific marker for kupffer cells so if any one 
knows please most welcome.
Thanks in anticipation.
 

Nasir Abbas
---PhD FellowGuangzhou Institutes of Biomedicine and HealthChinese Academy of 
Sciences190 Kai Yuan Avenue Science Park Guangzhou, 510530, Guangdong 
P.R.ChinaCell: +86-1312 8296 614
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