Re: [Histonet] Faded H tissue section

or the reagents, it should work just fine. Amos Brooks Message: 6 Date: Thu, 9 Nov 2023 22:02:46 +0530 From: "jayalakshmy p.s" To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Faded H tissue section Message-ID: Content-Type: text/plain; charset="UTF-8" Hello, I wo

[Histonet] Re IF staining questions

Hi, Direct IiF (a fluorescent conjugated primary) is certainly easier, but there are some reasons one might prefer using an indirect method (using a conjugated secondary). Indirect methods allow the use of a different wavelength to be used simply by switching the secondary. It is also

[Histonet] p17 mice brain sections (Alonso Mart?nez Canabal)

cutting them and transferring them directly to an 8 well plate with PBS and doing the IHC as a floating section. Cheers, Amos Brooks > > Message: 1 > Date: Tue, 19 Sep 2023 19:07:04 -0600 > From: Alonso Mart?nez Canabal > To: Histonet > Subject: [Histonet] p17 mice brain sec

[Histonet] Detection Systems

info is really frustrating. It has it's uses on occasion. Amos Brooks > -- > Message: 2 > Date: Tue, 1 Feb 2022 23:06:57 + > From: "Mac Donald, Jennifer" > To: "histonet@lists.utsouthwestern.edu" > > Subject:

Re: [Histonet] Microwaving Slides

Hi Samantha, Microwaves are terrible! I am really not a fan of them in general, but especially for drying slides, and even moreso for slides intended for IHC. There is no way to really monitor the temperature the slides get to. Sure you can get a fnacy one with a probe, but that probe

Re: [Histonet] Jones problems

ilver solution and on the slides be due to the > instability of the silver solution? > > > > Thank you so much! > > Jordan > > > > *From:* Amos Brooks > *Sent:* Saturday, September 25, 2021 8:03 AM > *To:* Hood, Jordan ; > histonet@lists.utsouthwestern.edu > *Subject:*

[Histonet] Jones problems

this helps, Amos Brooks ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Re: [Histonet] Histonet Digest, Vol 194, Issue 15

Hi Betsy, I wouldn't do it. It's an unnecessary risk to let them dry out. Better to leave it in distilled water if you absolutely must. Ideal to not deparaffinize in the first place if you can't finish the stain. Dried out sections is just a bad plan. Amos Brooks On Mon, Jan 20, 2020, 1:00

[Histonet] X-gal staining

> > > Hi, I'd like to concur with Carl Hobbs. With formalin fixed, paraffin embedded tissue, beta Galactosidase is definitely the way to go. It is just an antibody so you would do it like any other IHC. Amos Hi > No replies so far so.my pennyworth. > Imho ...no > beta Gal enzyme is

[Histonet] plural fluid prep

r you let it sit though, the more complications can occur like bacterial growth and further call division or death depending on what's going on in the fluid. Amos Brooks > > Message: 7 > Date: Wed, 18 Dec 2019 10:51:40 -0700 > From: Michelle Jamison > To: "histonet@lists.utsouthwest

Re: [Histonet] guidelines for using a microwave designated for food, but used to heat up histogel

> > Hi, There is NO reason to have a microwave in a histology lab. They don't really save any time and cause more problems than they are worth. You won't have to worry about any certifying agencies rulings about microwaves if they aren't there! Just put the Histogel in a rack in a

[Histonet] Tunel

Hi, > I hand stain this. It is a finicky test and benefits from a personal touch. When I have a ton of them, sometimes I do the enzyme on the stainer and then take it off for the primary and secondary then DAB on the stainer. Amos Message: 1 Date: Fri, 19 Oct 2018 19:24:01 + From:

Re: [Histonet] Histonet Digest, Vol 179, Issue 5

Hi, > I have a hunch that the DAB deposits you are seeing are not a DAB problem. You didn't mention how long your incubation time is. If I were to venture a guess, I am thinking there may be some evaporation of either the primary or secondary antibody or boil-over from the antigen retrieval.

[Histonet] Oil Red O

Hi, I have seen this too. I mitigate the problem by making it up in 50 ml increments and staining them flat. I draw from around the middle of the Falcon tube I make it up in. It tends to precipitate so you could filter it. Don't bother re-using the reagent. It's cheap & easy to make up. I

[Histonet] Science Haiku

Anyone that heard Science Friday recently may have heard of this. I thought it would be fun to see what others come up with. So many people think I'm an archaeologist when I tell them I'm a Histotech. This makes for a fun elevator pitch for our profession. Scientists are challenged to summarize

[Histonet] Tunel with Apoptag

Hi, I am contentedly using the Millipore (Chemicon) Apoptag IHC kit for Tunel stains. It works great... *BUT* I have been getting requests for a fluorescent tag rather than a chromogenic. The contents of the fluorescent kits are identical except for the anti-Dig. In the chromogenic kit is HRP

[Histonet] buffered zn-formalin recipe needed

Hi Johanna, The Histonet Archives are your friend. In 2009, Gayle Callis posted a great formulation that being the neo-Luddite that I am I printed and have a copy of here for just such an occasion... http://lists.utsouthwestern.edu/mailman/htdig/histonet/2009-October/046985.html Zinc

Re: [Histonet] Modified Pentachrome

e your safran du gatinais/saffron from? At this point I'm ready to start growing my own Crocus sativus. On Thu, Sep 1, 2016 at 12:35 PM, Amos Brooks <amosbro...@gmail.com> wrote: > Hi, > I use Lugol's Iodine all the time for this. It works just fine. I do > purchase certain ch

[Histonet] Modified Pentachrome

Hi, I use Lugol's Iodine all the time for this. It works just fine. I do purchase certain chemicals from manufacturers like Lugol's Iodine which I get from EMS. Most of the chemicals I make up myself from powder though. I have shared the procedure with you via Google Docs. I hope it helps.

[Histonet] Crystals in PAS

Hi, If you are having trouble with the instrument and the manufacturer isn't able to fix it, just hand stain them. PAS is really not a complicated stain. Please don't just accept underperforming equipment when we are all capable of so much more. Amos

[Histonet] Mucin on PR

Hi, I'm asking this for a colleague that is experiencing this problem on formalin fixed paraffin embedded human clinical samples. She is using PR636 from Dako on a Leica Bond platform. Has anyone seen mucin or any cytoplasmic staining with Progesterone receptor antibody? Thanks folks, Amos

[Histonet] PPE/ Hair Regulations

If it is actually getting caught in things or falling in waterbaths or embedding centers and such, then it needs to be tied back. The same is true of food service. You can't risk contamination. If it is under control though, leave it alone. Don't try to use safety to enforce your personal bias of

[Histonet] Tissue Arrays

Hi, My experience with microarrays is that they are sometimes a bit complicated to work with depending on the way they are constructed and the platform they are used on. Often these slides are dipped in paraffin to help preserve them. When this happens they need considerably longer

Re: [Histonet] IHC with H & E staining

Hi, I usually try to avoid eosin as a counterstain for a DAB labeled slide because the red/pink of the eosin can obscure the rusty brown of DAB. If you really want to use it though I would suggest a *really* light eosin, perhaps even just a few milliliters in the 95% ETOH as you are

[Histonet] GATA3

Hi Lisa, Skin seemed to work fine for me. Amos On Tue, Dec 22, 2015 at 1:00 PM, wrote: > Message: 2 > Date: Mon, 21 Dec 2015 13:12:14 -0500 > From: "White, Lisa M." > To: > Subject: [Histonet]

[Histonet] VEGF

Hi, VEGF is driving me nuts. I was hoping someone might have some suggestions. I have been using clone VG1 for a while. I have never really been particularly happy with it. The labeling is not nearly as specific as I would like and there is almost always some background. If I dilute it more

[Histonet] Purple elastic

Hi, There is a reason that every manual with an elastic procedure says that the stain should be made up fresh. My guess is that the company is trying to prolong the shelf life by removing or reducing either the iodine or ferric chloride. Doing this will prevent the hematoxylin from turning

[Histonet] H2O2 blocking

Hi Gayle, Thank you again for the insight. You are a wealth of knowledge. I am also not particularly surprised that I was incorrect in my assumption about Peroxabolish. I really like to know what is in the products I am using. That's why I prefer to make up my own reagents whenever possible,

[Histonet] More on H202 issues

Hi, Peroxidase can really be a pain. If you look in the archives though (or ask her really nice) Gayle Callis submitted a recipe for a glucose oxidase for peroxidase quenching that does not include hydrogen peroxide. If you aren't really a fan of making these things up I would bet dimes to

Re: [Histonet] training log

Hi, Actually that would be cool for comparison purposes for me too. Perhaps it might be easiest to put them on dropbox and post a link to the histonet. But please share. Thanks, Amos Brooks On Tue, May 19, 2015 at 1:00 PM, histonet-requ...@lists.utsouthwestern.edu wrote: Message: 12

[Histonet] B-gal positive control

Hi, Normal kidney should work fine for this. Amos On Fri, May 8, 2015 at 1:00 PM, histonet-requ...@lists.utsouthwestern.edu wrote: Message: 9 Date: Fri, 8 May 2015 14:50:42 + From: Coffey, Anna (NIH/NCI) [C] anna.cof...@nih.gov To: histonet@lists.utsouthwestern.edu

[Histonet] Tissue transfer method:

Hi, I have used a product called Mount Quick. It is designed as a liquid coverslip, but once applied, it can be removed lifting the sections along with it. Here is a link with a good description of the process. http://www.newcomersupply.com/product/mount-quick Happy Friday, Amos Brooks

[Histonet] H. Pylori Testing

into a catch tray below. It makes it **really** easy to reproducibly do IHC manually. Amos Brooks On Tue, Apr 28, 2015 at 12:24 PM, histonet-requ...@lists.utsouthwestern.edu wrote: Message: 8 Date: Mon, 27 Apr 2015 16:49:25 -0400 From: Garreyf garr...@gmail.com Subject: Re: [Histonet] H

[Histonet] setting up for staining

Hi, It's a good setup for a smaller volume lab. The racks hold 10 slides and the dishes slide together and link into a chain or can be used separately. I have used the dishes for antigen retrieval and they haven't cracked or warped. I haven't regretted the purchase. Cheers, Amos Brooks

[Histonet] type IV collagen antibody

Hi, The one from AbCam Cat# ab6586 works nicely. I have also used Dako's Coll IV with some success. Amos On Wed, Apr 8, 2015 at 1:00 PM, histonet-requ...@lists.utsouthwestern.edu wrote: Message: 4 Date: Tue, 7 Apr 2015 14:32:58 -0700 From: Amy Lee amylee...@gmail.com Subject:

[Histonet] Trichrome Fixation

Hi, It is interesting that you should mention the importance of fixation on the Trichrome stain. I have an image of two murine hearts processed, cut and stained side by side. The only difference between the two is that they were harvested at different times, so one sat in formalin long enough

[Histonet] Epitomics

Hi, You will want to contact AbCam. They bought Epitomics a while ago. Their website is www.abcam.com and their support is great. Amos Message: 7 Date: Thu, 15 Jan 2015 16:50:47 -0500 From: Clare Thornton cthorn...@dahlchase.com Subject: [Histonet] Epitomics To:

[Histonet] Slides for IHC

Hi, Part of checking out slides for use should be to take a *blank* slide (right out of the box with no section on it) and look at it under a fluorescent microscope. With increased FISH and fluorescent labelling lately, we have seen many slides that have fluorescent inclusions within the

[Histonet] BrDU

Hi, I use Sigma's B2531 with 1 hr pretreatment in 1N HCl at 40 deg (in a coplin jar in my waterbath) then 15 min Trypsin (don't forget the CaCl2) at 40 deg. I detect it witb Envision+ but any mouse IgG would work fine. Amos Message: 15 Date: Tue, 24 Jun 2014 15:12:01 + From: Chiriboga,

[Histonet] Re: Histonet Digest, Vol 127, Issue 3

Hi Alpha ;-), I usually tell new techs that you will not find and should not look for the perfect job right out of school. It is imperative that you get yourself a broad base of experience before settling down with something you really like. Eventually you will find your niche. You have

[Histonet] Re: Histonet Digest, Vol 126, Issue 15

Hi Andi, I have a Google Doc of our procedure. I just sent you an invite to view it. If anyone else wants to see it, just let me know I'll share :-) I know the procedure calls for overnight in the LFB solution. It's for the best, the results are much better if you are patient. It is really

[Histonet] Leica IPs slide printer

Hi, Leica instruments are usuly over-engineered finickey pains in the neck that work fantastic when they want to but really want to try to fly out the window with troubling regularity. Our IPs has been babied for a long time. The ink on these tends to get really gummy after a while. You

[Histonet] (no subject)

Hi Tresa, I haven't seen any responses to this question, so I figure I should take a crack at it. You are right that using a polymer detection system is so clean that you hardly have much to worry about background/non-specific staining when not using a buffer for wash steps. There is

[Histonet] Mouse GranzymeB

Ok Musketeers, I am trying to detect cytotoxic T-Cells in formalin fixed paraffin embedded mouse liver. GranzymeB should do the trick. I have a rabbit anti mouse (and human and rat) GranzymeB from abCam (ab53097). The spleen that I ran as a control with it works fine. Nice T-cells and no

[Histonet] CAP Survey Question

Hi, Regarding the question of weather it would save time to not have the techs QC check the IHC slides. I feel that it would be an extremely bad practice to omit the techs from the QC process. This goes for not only IHC but special stains and even HE stains. It is imperative that the techs

[Histonet] (no subject)

Hi, There are a few ways of doing this. Perhaps the easiest is scraping the cells off while it is still in the media. Transfer it to a centrifuge tube and spin it down. Pour off the supernatant add formalism vortex it and centrifuge it again and pour off the supernatant again and add

Re: [Histonet] PicroSirius Red in Frozen Sections

Peggy, That was a fantastic answer and it is responses like that to such questions that is the whole reason I enjoy the Histonet so much. Thank you for such a well thought out answer. Amos On Tue, Nov 12, 2013 at 1:00 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 7 Date:

[Histonet] Speaking of picrosirius red...

Hi, In light of the current discussion about picrosirius red I would like to revisit a question that was asked a while back. If it was answered publicly, I apologize I probably missed it and didn't see it in the archives. Someone asked what is the purpose of the phosphomolybdic acid step

[Histonet] Manual IHC

an instrument is certainly justified. Best of luck, Amos Brooks ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Unsubscribe, Chapter 195

Hi, I do not disagree at all. in fact, I think it would make for an interesting management tool. If you have the nicest CV around, but a simple Histonet archive search for your name + unsubscribe shows that you can't follow simple instructions, it says something about your communication

[Histonet] Re: Histonet Digest, Vol 118, Issue 2

Hi Laurie, You can pick them up from Home Depot or Lowes. They are mini scrapers. Amos On Tue, Sep 3, 2013 at 1:00 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 2 Date: Tue, 3 Sep 2013 14:16:13 + From: Laurie Colbert lcolb...@pathmdlabs.com Subject: [Histonet] Razor

[Histonet] HSP70

Hi, We had decent luck with the abCam antibody ab5542 at 1:1600 with no retrieval. Good luck, Amos ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet

[Histonet] Re: Histonet Digest, Vol 113, Issue 10

hauler in the first place. Ultimately, the decision should be made by your Department of Environmental Protection. It would be best to share the MSDS directions for use with them and let them decide if it is a good idea before trying to use it. Amos Brooks On Thu, Apr 11, 2013 at 1:00 PM

[Histonet] Stain to ID eosinophils specifically

Hi Colleen, Although I was not able to locate the specific reference you listed, the two stains that come to mind are Lendrum's Carbol Chromotrope and Congo Red. While they are not *really* specific for eosinophils, they do stain eosinophils well. It is fairly easy to differentiate an

[Histonet] Temperatures

: WILLIAM DESALVO wdesalvo@outlook.com Subject: RE: [Histonet] Temperatures To: Amos Brooks amosbro...@gmail.com, histonet histonet@lists.utsouthwestern.edu Message-ID: bay002-w158efe2f3accfae9c9a033c82...@phx.gbl Content-Type: text/plain; charset=iso-8859-1 Who cares? The patient

[Histonet] Temperatures

Good Grief! Why would this really be an issue. The temperatures are taken throughout the week and are constant (or you have a different problem entirely) why would they only spike or tank on the weekend, and why would it even matter if the equipment isn't being used. If a tree falls in the

[Histonet] Re: Histonet Digest, Vol 110, Issue 25

. Best of luck, Amos Brooks On Sat, Jan 19, 2013 at 1:00 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 3 Date: Fri, 18 Jan 2013 19:51:20 + From: Herrick, James L. (Jim) herrick.ja...@mayo.edu Subject: [Histonet] SRBS/van Gieson To: histonet@lists.utsouthwestern.edu

[Histonet] Trichrome

. One final test could be to make up fresh solutions and try again, although I would suspect you are likely to get more of the same. All the best, Amos Brooks On Tue, Dec 4, 2012 at 1:00 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 3 Date: Tue, 4 Dec 2012 05:18:35 -0800 (PST

[Histonet] Problem with cardiomyocytes staining

Hi Laura, I do this stain very frequently. It can be finickey. You should make very sure the pH is below 2.5. It could be that you have an old solution and over time these tend to drift toward a neutral pH. This will affect the staining. Sometimes the solutions need to be discarded and either

[Histonet] Picric acid

Hi, The Halifax explosion was indeed a very dramatic event. Anyone unfamiliar with the story should certainly read up on it. It was truly incredible. The link to the Wikipedia article was previously posted. A couple of important points about this story. The ship that blew up was carrying metric

RE: [Histonet] air drying special stain slides rather than dehydrate and clear

...@lists.utsouthwestern.edu] On Behalf Of Amos Brooks Sent: Tuesday, September 11, 2012 7:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] air drying special stain slides rather than dehydrate and clear Hi, My choice to air dry rather than dehydrate in ETOH xylene is based

[Histonet] air drying special stain slides rather than dehydrate and clear

Hi, My choice to air dry rather than dehydrate in ETOH xylene is based on the stain rather than the spooky xylene hazard boogyman. Yes, not using xylene if it is not really needed is not a bad idea, but the main reason I air dry some stains is the alcohols remove some of the stains. Ever

[Histonet] endothelial cell marker

Hi, My favorite by far is VonWillebrandt (spelling?) factor VIII. Dako has a really good one (Cat# A0082) that works great in mouse tissue. CD31 is OK but it is really finicky. CD34 works but F8 is easier and more reliable. Amos On Wed, Sep 5, 2012 at 1:01 PM,

[Histonet] calcium buildup

build up there and can actually clog the drain if left unchecked. Actually this should all be part of a periodic maintenance plan. We have a service take care of it for us (Belair in New Jersey comes out for us) and I also take it down and clean it out before our Christmas break. Amos Brooks

[Histonet] counterstain for Alcian Blue (ph2.5)

Hi, We use Neutral Red in 1% solution. (Not being in the lab right now I think it is made up in 1% acetic acid, but I could be wrong.) It is much faster than NFR (nucear fast red). It is about one minute to stain as opposed to the 5 to 15 if the NFR solution is old. It is easy as heck to make

[Histonet] Golgi-Cox Sections

Hi, For those that nave had the joy of performing the Golgi-Cox stain, you will likely recall that it generally calls for some thick sections. The researcher I am working with is hoping for 100 uM sections. That is way too thick for a microtome and a cryostat. I suggested a vibratome might

[Histonet] Elastic Stain

Hi Dorothy, Controls for Verhoeff vanGieson elastic stain is very subjective due to the differentiation in ferric chloride. Large arteries are usually used as a control. If you differentiate all the slides based upon the appearance of a large artery the small vessels will be totally gone. If

[Histonet] BLock alignment tool

Hi, There is one on every microtome made. The two little knobs align the block just perfectly to any angle previously cut. It is much better than just whacking into a block that was cut slightly differently than what a dumb instrument is telling you is aligned. Knowing how to use this

[Histonet] Embedding

Hi, Any chance it is from your reservoir? Sometimes the paraffin is not filtered very well and there is some junk that collects in the chamber. (Yes I'm looking at you Paraplast!) Also is the reservoir you keep your cassettes in prior to embedding clean? Perhaps junk on your tampers? Amos

[Histonet] Re: Histonet Digest, Vol 102, Issue 28

Hi, I have done this. It works quite well. It would probably be best to use an alkaline phosphatase detection since the brilliant red of the fast red precipitate is more distinguishable from the brown DAB. (I am assuming you used HRP-DAB detection of the Tunel.) You needn't worry about the left

[Histonet] DeCloaking Chamber

Hi, Here are the best steamers on the market... http://www.walmart.com/ip/Black-Decker-7-Quart-Food-Steamer/14320967?findingMethod=rr There are others that will work as well. Use only one layer. The second layer is never as hot as the first. I drill a hole in the top and drop in a

[Histonet] Problems with pAKT.

Hi Courtney, We tried it and didn't have much luck. I think this antibody is in need of a lot more development. We abandoned pAKT for panAKT from Epitomics. I am still not 100% content with any of the AKT antibodies, but this was the best we tried. Good Luck, Amos On Wed, Apr 25, 2012 at

[Histonet] Re: Histonet Digest, Vol 101, Issue 27

Hi, Strange issue with your Ki-67. I don't know what would do that unless it was non-specific background. Is it nuclear or cutoplasmic? If you are working in mice or other animals you could try labeling them with BRDu then detecting it with an anti BRDu antibody. Humans don't like being

[Histonet] Re: placenta encapsulation

Hi, I would recommend getting someone else to do it for you. Unless you are *really* resilient, I would expect you would have other recovery related things to think about. With the OB/GYN's permission (and curiosity) I took my wife's placenta twice. Once for each kid. The local college was

[Histonet] Cardiac Myocytes

Hi, What would one use to identify cardiac myocytes in mice. I know there are specific antibodies, but I was kinda hoping for something a bit more mundane like desmin. Any ideas? Amos ___ Histonet mailing list Histonet@lists.utsouthwestern.edu

[Histonet] fibrin IHC on FFPE mouse tissue

Hi Kim, I am working on a project just like this right now and we are using Fibrinogen (AbCam ab34269) and PTAH. Drop me a line if you have any questions. Amos On Mon, Feb 27, 2012 at 1:00 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 20 Date: Mon, 27 Feb 2012 09:34:08

[Histonet] Cytology CSF Cell Pellets made from Histogel

Hi, I haven't done a cellblock on CSF with Histogel, but I have has success with some fairly scanty cell culture specimens. A short processing cycle would be best. Try to make sure you have removed as much supernatant as possible to keep the gel from shriveling during processing. The IHC will

[Histonet] Music in the lab

Hi, I am in the same boat as those that would go nuts without music. I have noticed that I cut a *heck* of a lot faster with music than without. It's a rhythm thing, I can't really explain it. I really can sympathize with those that can't tolerate certain types of music though. Having one tech

[Histonet] Formalin Neutralizing

Hi, To take it a step further, if you look at the MSDS of the Neutralex, it is very nebulous as to what the chemical is. We couldn't dump the Neutralex alone down the sink since we don't know what it is. So combining a hazardous chemical with an unknown chemical then dumping it down the sink...

[Histonet] CD marker help [particularly CD68]

Hi, It sounds like there are a number of antibodies with similar problems here if you are having troubles with CD68, F4/80 and CD11b. If you are getting variable results it could be that the tissues are not fixed and processed sufficiently. If they are not fixed sufficiently there will be

[Histonet] Nerve Fiber Density Testing

Hi, I did this same thing some years back. Keeping the free floating skin bx sections from breaking up during the IHC was tricky. Sometimes it is easiest to transfer the solutions in out of the plate wells rather than trying to lift the sections with a loop. It can end up with good results

Re: [Histonet] b-cells stain

alum hematoxylin with either potassium dichromate or ferric chloride! Cheers, John Kiernan Anatomy, UWO London, Canada = = = On 01/12/11, *Amos Brooks *amosbro...@gmail.com wrote: Hi, I like to prepare solutions myself as well. As you say though paraldehyde is classified as a drug so

[Histonet] b-cells stain

Hi, I like to prepare solutions myself as well. As you say though paraldehyde is classified as a drug so there are hoops to jump through to be able to purchase it. This is one of those times that it totally makes sense to let someone else make it up. EMS sells aldehyde fuchsin so it saves me

[Histonet] validation

Hi, I would say no, and if any inspector disagreed you would be well within your rights to give him a good cuff upside the head. The point in having a negative mouse and rabbit is to run them at the same concentration as whatever primary you are running. That would mean you would need to

[Histonet] microtomes

Hi, If you are concerned about the thickness of the sections being accurate to the setting, I would suggest picking up a cheap micrometer from a hardware store. (OK perhaps not cheap as you will want a quality one, but the cost of these isn't terrible.) You can't really measure 5 microns (or

[Histonet] Immune cells antibody

Hi, You can detect NKT cells with CD57. They are also labelled with Granzyme B too, but I think CD57 will be a bit more specific. Amos On Tue, Nov 8, 2011 at 12:32 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 12 Date: Mon, 7 Nov 2011 09:48:01 +0100 (CET) From: Carmen Maria

[Histonet] Stripping antibodies

Hi, Stripping sites are usually found in the more seedy areas of large cities ... Oh wait you meant ... nevermind :-) If you developed the reaction with DAB, you may be in a difficult position. DAB is a really strong reaction. You might try a Mallory bleach to remove it. 0.3% sulfuric acid in

[Histonet] Annexin V IHC staining in mouse xenograft

Hi, Have you considered running the antibody through some antibody purification beads. It isn't really a common histolgy thing to do, but the process isn't terribly difficult. It really sounds like a dirty antibody is at fault. It will probably result in a cleaner and more concentrated

[Histonet] Von Kossa stain kit

Kits ... Bah! Just make it up. Save yourself a fortune and actually know what's in the solutions. All the classical texts should have the procedures as well as various websites like stainsfile.org IHCworld.com Otherwise just ask here and I'm sure someone will have the information you need

[Histonet] Paraffin sections for molecular assays

Hi, You will want to do more than just wiping it down. You should use something specifically designed to clean such contamination like RNAase away (Invitrogen if memory serves). You'll want to wipe down any surfaces that come in contact with the block, sections or your hands when sectioning.

[Histonet] Atlas of laboratory mouse histology

Hi, I would be interested in this reference as well. Please post to the group if you have any suggestions. Amos On Mon, Oct 3, 2011 at 1:00 PM, histonet-requ...@lists.utsouthwestern.eduwrote: Message: 10 Date: Mon, 3 Oct 2011 11:40:55 -0400 From: Hinsinger Julie julie.hinsin...@umontreal.ca

[Histonet] Xylene sensitivity

Hi, I can attest to the effacacy of the isopropyl/mineral oil processing that Rene describes. It works great for murine heart samples. I would not switch entirely though, since xylene does a fine job with most other purposes. My main concern is the rather hysterical impulse to get rid of every

[Histonet] AO-860 Sliding Microtome + OCT Embedded Tissue?

Hi, Got a walk in freezer? Really, cutting large slabs of OCT embedded material is just not possible on a sliding microtome unless you keep that microtome in a -20 freezer. You could cut small blocks on it by mounting the OCT on a large chuck and surrounding it with dry ice. This will really

[Histonet] Question of combining immunofluorescence and more conventional cytologic stains

Hi, Have you tried doing the conventional stain first, then the fluorescent after. As long as the epitope survives the initial staining, it should be fairly easy to label the cells with a fluorescent tag. Amos On Sat, Sep 17, 2011 at 1:00 PM, histonet-requ...@lists.utsouthwestern.eduwrote:

[Histonet] Von Kossa vs Alizarin Red

Hi, I do both often. You would want the VonKossa rather than Alizarin Red if you had any plans to do any subsequent double staining. The silver isn't going anywhere. The Alizarin Red will disappear as soon as you place the slides in water (or alcohol) so you can't stain anything else. Since

[Histonet] immunohistochemistry: manual staining

Hi, Shandon Sequenza staining racks and slide clips. They are great. You don't need the whole system (timers boxes to hold the reagents). Basically this is a rack that holds slides and slide clips vertically and the reagents are dropped in at the top of the slide and it displaces the

Re: [Histonet] immunohistochemistry: manual staining

...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Amos Brooks Sent: Tuesday, September 13, 2011 3:34 PM To: histonet@lists.utsouthwestern.edu; wim.vandenbro...@ugent.be Subject: [Histonet] immunohistochemistry: manual staining Hi, Shandon Sequenza staining

[Histonet] Digital Security

Hi, When this happens, first change your email password. That may be the best way to prevent further breaches. Next scan your PC (certainly not a Mac or linux right?) with a good antivirus program. AVG and Clamwin are both decent free antivirus programs. Next it would be good to scan for

[Histonet] Histonet] Re: Embedding process etc.

Hi, That was a good article. I really do hope I am not the only one that was just mortified at the high numbers of bad sections being produced. Once in a while on a bit of difficult tissue is one thing, but consistent poor quality is inexcusable. Come to think of it that was actually a rather

[Histonet] Immuno. confusion

Hi, Well... almost any point before DAB. You wouldn't want to put it in after the HRP conjugated secondary because that would quench the HRP signal you are actually looking for leaving you nothing for the DAB to precipitate on. On the other hand you certainly do not *need* to do it after the

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