[Histonet] Best second chromogen with DAB IHC?

Hi there, we want to do some double labelling, one of the chromogens being DAB and haematoxylin the counter stain. Which chromogen would you choose for the second labelling? We use envision-HRP as secondary As always, thanks a lot for your help! Cheers julio Julio Benavides Silván

[Histonet] comparison between microtomes leica RM 2125RTS y thermo HM325

Hi there, we are buying a new microtome for the lab (research, not high load of work) and have been offered leica RM 2125RTS and thermo HM325 with similar prices. Could anyone help us to decide? thanks a lot cheers Julio ___ Histonet mailing

[Histonet] Update on Trimming soft brain

in alcoholic formalin did not interfere with the IHCs, at least with the one we are using (CD3, bAPP and GFAP). Again, thank you so much for your help! cheers Julio El 06/03/2017 a las 23:18, Julio Benavides via Histonet escribió: Hi there, I´m trying to trim ovine foetal brain (90 days

Re: [Histonet] Sections falling off

Just to add another advice, Pablo, yesterday I told you that we were drying the slides overnight at 37º... not true!! (that happens for me speaking without checking with the really knowledgeable ones, the technicians!! We used to have that same problem and then began to dry at 60º for one

[Histonet] Trimming soft brain

Hi there, I´m trying to trim ovine foetal brain (90 days of gestation). The brain is too soft and there is no way to get a proper slice. It has been in buffered 10% formalin for three weeks but still too soft. When you take the brain out of the pot and put into the trimming table, it just kind

[Histonet] Leica DM2000 microscope + integrated camera

Hi there, We are considering buying a leica DM 2000 microscope with an integrated camera ICC50W. Anyone has any experience with this microscope? is the camera any good (thinking mainly on discussion sessions with students but also your odd good quality picture for publication). Your

[Histonet] Processing of frozen material

Hi there, I have frozen (-20ºC) samples from skeletal muscle (sheep samples) that now requiere processing and cutting for HE and IHC. Any advice on how to proceed? better to put them in formalin while still frozen or first thaw them? A good way to thaw them preserving the most of the

Re: [Histonet] Macrophage Stain

Hi Jessica, there are actually quite a number of Abs working on sheep tissues. Take a look at this paper, most of them work in sheep (I believe all but gamm-IFN). Let me know if you cannot access the paper and I will send you a copy. https://www.ncbi.nlm.nih.gov/pubmed/27315822 Happy to

[Histonet] Substituting formalin by 70%ethanol in fixed samples.

Hi there, This is me again with formalin issues. The health and safety officer of my institute keeps on in her crusade of eliminating formalin from the world. After your helpful emails, I convinced her that you do need formalin to fix samples, other approach would need to much setup

Re: [Histonet] veterinary histology

Hi there, could be E. coli+virus ("swallen head syndrome") or fowl cholera (P. multocida). I would check the nasal cavity (slice the peak longitudinally) and also the infraorbital sinuses. I am at aloss regarding the ear discharge. Try a coronal slice of the cranium at the ear opening. A

Re: [Histonet] Formaldehyde and Xylene Detector

I would also be very grateful if you could help me with this issue. We are trying to monitor xylene and formaldehyde levels in our lab, but we cannot find adequate measurement devices. What do you use? is there any equipment capable of give you an envoiromental level of these products in the

[Histonet] Stamp staining in paraffin and smear samples

Hi There, Is anyone using this technique to stain chlamydia in placental samples (working with sheep here)? I was looking for a working technique but it looks like it is not easy to find. Thanks for your thoughts! Julio Rene J Buesa via Histonet

Re: [Histonet] picric acid

2 19. Documentation of the Threshold Limit Values, American Conference of Governmental Industrial Hygienists, Inc.: Cincinnati, Ohio, 1991, page 1271. 06/18/02 +++++++ -Original Message- From: Julio Benavides via Histonet [mailto:histonet@lists.utsouthwe

Re: [Histonet] picric acid

Hi, For how long can you keep it in water? any particular dilution or just keep it humid (saturation)? We also do have some dry picric acid in the lab and, after reading about the bomb squad, I was begining to get concerned... Thanks a lot julio El 06/05/2016 a las 15:30, Rene J Buesa

Re: [Histonet] Mycoplasma

No, it won´t. You better think on IHC for labelling mycoplasma. cheers Julio El 05/04/2016 a las 20:12, Bernice Frederick via Histonet escribió: Hello all, Will PAS stain mycoplasma? Bernice Bernice Frederick HTL (ASCP) Senior Research Tech Pathology Core Facility Robert. H. Lurie Cancer

Re: [Histonet] Image analysis software

Hi Jodi, I was using imageAnalysis from Olympus some years ago, bus since I discovered ImageJ there is no way I will go back to commercial stuff. Pros of ImageJ, apart from the obvious one that it is free, is that a lot of people is using it, so plenty of tutorials, tricks, how-to in

[Histonet] Collodian bag for cell blocks

Hi, I have a question regarding the use of UCSF Collodion Bag Cell blocks. once you do the bag, you pour in the cells in, let´s say 10ml of formalin. Then, how do you proceed? Do you spin down the cells with the bag inside a plastic tube? do you take then put the formalin and cut the top

Re: [Histonet] Davidson's fixative and IHC question

Hi, in my experience, Bouin´s fixative can sometimes give you problems with IHC. Both because it seriously interfere with some antigens (in my case, I was doing IHC for lentivirus and herpesvirus and more than 24h of fixation would drastically reduce IHC labelling) but also because it

[Histonet] Pinkerton's adaptation of machiavello's stain for chlamydophila on tissue sections

Hi there, I was wondering if any of you had used the Pinkerton's adaptation of machiavello's stain for staining chlamydophila on parafin tissue sections and whether it works or not. I would also be very grateful if anybody can advise in any other technique working on tissue sections. We

[Histonet] Trypsin vs. proteinase in IHC

Hi everybody, I was wondering whether you can use proteinase K instead of trypsin for antigen retrieval in immunohistochemistry. Sure thing you can use both, but do they unmask the same antigens? Thanks a lot for your thoughts, All the best Julio

[Histonet] Eosinophil labelling

Hi everyone, Is there anti antibody , working on FFPE, labelling eosinophils? if it works in sheep then that´s perfect! I have seen several histochemical methods (Modified Congo Red, Luna Protocol, Astra Blue/Vital New Red Protocol (AB/VNR, Sirius Red Stai ). Does anyone has any experience

[Histonet] Opinion on PTlink from Dako

Hi, we are thinking about incorporating the PTlink system, from Dako, in our lab. Have any of you worked with it before? Any opinion? They would be greatly appreciated Thanks a lot Cheers Julio University of León Spain ___ Histonet mailing list

[Histonet] Reagents changing in dewax/dehydration and tissue processor

Hi histonets, how often do you change alcohol/xylene for the dewax-dehydratation? we are running a manual system, with racks of 20 slides in 200 ml containers. These is the routine: *Dewax:* Xylene Xylene 100 ethanol 96 ethanol 60 ethanol *Dehydrate:* 96 ethanol 100 ethanol 100 ethanol