[Histonet] mouse femur decal and paraffin embedding protocol

Dear Histonet Colleagues, I hope the holiday season has been good to all. We are interested in a routinely used protocol to decal, process and paraffin embed mouse femur and tibia. Some of our projects may require a gentle decal, whereas others may not be so delicate. Also, we do have access

[Histonet] Issues with rat mandible sectioning and staining

Good morning Everyone!! We have embedded several rat mandibles in MMA (we add Perkadox to the MMA to initiate) but are having issues with sectioning. We would like to keep the sections close to a 5 um thickness and need to stain with an HE. Unfortunately, we are having issues with sectioning

[Histonet] Question on Fluorochromes for Histomorphometry

Dear Histonet Colleagues, I hope everyone is doing well-- We are going to be using a goat mandible model (embedded in MMA) for a project and will need to administer tetracycline and/or calcein (others if suggested) for histomorphometric analysis. Would anyone be kind enough to share their

[Histonet] SRBS/van Gieson

Happy Friday Everybody, We are trying to develop a protocol for an SRBS/van Gieson's stain on MMA embedded tissue. The sections are 5um in thickness and deplasticized. Our protocol seems to work ok on thick sections but not on thin, deplasticized sections. Any thoughts or ideas would be

FW: [Histonet] iliac artery attachment to slides?

Good morning everybody!! I am trying to attach iliac arteries embedded in MMA (methyl methacrylate + dibutyl phthalate + perkadox 16) to gelatin coated slides to allow us to stain them with the Movat's Pentachrome. Unfortunately, I have a real problem getting them to attach - shortly after they

[Histonet] iliac artery attachment to slides?

Good morning everybody!! I am trying to attach iliac arteries embedded in MMA (methyl methacrylate + dibutyl phthalate + perkadox 16) to gelatin coated slides to allow us to stain them with the Movat's Pentachrome. Unfortunately, I have a real problem getting them to attach - shortly after they

[Histonet] Movat's Pentachrome Question

Happy Friday everyone!! I am trying to find a good Modified Movat's pentachrome stain protocol for plastic embedded iliac arteries and also bone tissue (animal model). Everything I have been able to find so far, is for paraffin embedded tissue and a mile long. There may not be a fix for the

[Histonet] Safranin O

Hello everyone, I am trying to stain for cartilage in mouse skull (cross-sectioning with the brain in tact) and wanted to know if anyone might have a good protocol for Safranin O / Fast green or Safranin O / von Kossa? The tissue is embedded in MMA and was cut on a saw microtome and finished down

[Histonet] Plastic discoloration

Good morning everyone, I just have a quick question in regards to GMA turning white following embedding. Does anyone have any experience on why our plastic is losing its clarity and turning a whitish color? This doesn't seem to have any impact on staining but it would be nice to know the reason.

RE: [Histonet] Problems with histonet

I made a post this morning as well, and it didn't show. However, I did receive a response. Jim -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Akemi Allison Sent: Friday, December 03, 2010 6:54 PM To:

FW: [Histonet] Liver Processing Question

-Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Herrick, James L. (Jim) Sent: Friday, September 10, 2010 2:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Liver Processing Question Hello

[Histonet] Liver Processing Question

Hello everyone, I am trying to process and embed liver tissue in MMA. I used the same protocol that has worked very well a hundred times over, for harder tissues, but when used on the liver tissue, polymerized into a relatively soft and rubbery textured plastic for one of the two specimens (the

[Histonet] Staining Question

Good afternoon everybody, Would anyone be willing to offer their wisdom on the best etching solution for 50 to 75 micron thick jaw specimens embedded in MMA? The stain we want to use is a Trichrome. Thanks in advance for your help. Jim ___ Histonet

FW: [Histonet] Mounting Medium Issues

-Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Herrick, James L. (Jim) Sent: Friday, May 14, 2010 3:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mounting Medium Issues Hello everyone

[Histonet] Mounting Medium Issues

Hello everyone, Hope you have all had a good week!! Has anyone had any negative experiences using Eukitt as your mounting media? We embed our specimens (human - iliac crest, animal - femurs, tibias, vertebrae, etc.) in GMA and MMA and have noticed over a period of time that a fairly large number

[Histonet] Processing Question

Dear Histonet Colleagues, I am trying to develop a processing protocol for our Leica TP1020, but the first run has not been very successful. The specimen is canine femur (section lengths of 1 to 1-1/2 ). Following is a list of the processing reagents and times for my first attempt:

[Histonet] Processing Question

Dear Histonet Colleagues, I am trying to develop a processing protocol for our Leica TP1020, but the first run has not been very successful. The specimen is canine femur (section lengths of 1 to 1-1/2 ). Following is a list of the processing reagents and times for my first attempt:

[Histonet] Question on Tissue Processing

Hello all, We have recently received a Leica TP1020 tissue processor and will be setting it up very soon. Our specimens will be embedded in MMA. Would anyone have any protocols that have worked well with rat and mouse femurs/tibias, as well as for larger specimens (e.g. goat and sheep spines,

[Histonet] MMA embedding issue

Hello everyone, I have embedded several rat femur/tibia specimens in MMA and am finding large holes (un-polymerized tissue) as I cut into the tissue - I am still in the soft tissue and not even into the bone yet (the specimens have large tumors). I placed a couple of the specimens into a 50°C

[Histonet] Question on decalcification of canine femur

Hello everyone, Would anyone have any experience with decalcifying canine femurs (approximately ¾ dia. X 1 length)? We are currently using a Ted Pella microwave with a 14% EDTA solution and a pH of 7.4. The microwave is on for approximately 7 to 8 hours daily (excluding weekends) and the

[Histonet] Section Mounting Techniques

Hi everyone, Does anyone have a good technique for mounting bone tissue (rabbit calvaria embedded in plastic) to coated slides? I am having problems with wrinkles following the rolling of the sections onto the slides. I would greatly appreciate any help you might be willing to offer. Thanks. Jim

[Histonet] Question on Sanderson's Rapid Bone Stain

Hello everyone, I am trying to run an SRBS on a 5 µm thick, deplasticized (MMA) section of rabbit calvaria. I have had really good luck with thick sections (plastic left on), but cannot get a good stain on the thin, deplasticized section. Does anyone have any protocols they don't mind sharing?