[Histonet] Microtomy of Equine Tendon Blocks

Gerard Spoelstra G.Spoelstra at murdoch.edu.au
Thu Dec 7 20:25:18 CST 2017

Hi Alan

We have had success with horse's hoof using potassium hydroxide prior to processing. After the hoof is fixed for several weeks we leave the hoof for 12 hours sometimes longer in 10% Potassium Hydroxide and then further trim the tissue in. The Potassium Hydroxide  has a marked affect in softening the tissue without affecting the staining.  The tissue is processed as usual. We all our process the tissue with our paraffin temperature set at 60 degrees C. This gives better infiltration of hard tissue. I would try with one block. Take it back to water leave it in NBF for O/N the leave it in 10% Potassium Hydroxide for 12 hours. After washing in water  process as usual.


Gerard Spoelstra

Veterinary Histology

Murdoch University

Perth Western Australia

From: Alan via Histonet <histonet at lists.utsouthwestern.edu>
Sent: Wednesday, 6 December 2017 12:03:07 AM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] Microtomy of Equine Tendon Blocks

Greetings to all on Histonet. I have a particularly challenging question regarding the sectioning of Equine tendon material. My colleague and I recently received a number of TS tendon slices. Unfortunately they had been placed in 90% IDA for fixation approximately 10 years ago, where they remained up to the point they were sent to our lab for cutting and staining.
The blocks are extremely hard, almost like undecalcified bone. They have processed well into paraffin, but we have wrecked several disposable blades just in facing the blocks. They have a very gritty feel on trimming even at 2 microns.
We have tried some commercial softening agents, including Mollifex, with long immersion over days, rather than the usual hour or so, we have tried neat fabric softener and 10% phenol in 70% IDA. We have also left some blocks in molten paraffin for several days, as this sometimes works with very fibrous tissues. As does prolonged chilling with ice and water.
We have managed to recover a small number of reasonable sections, at present we are looking at 1 block = 1 blade to obtain a very few working sections.
Do any histotechs out there have any secret recipes that they are prepared to share and have made up themselves, or a popular in-house reagent that they know is reliable,  that they have enjoyed success  with in their labs, or references to softening solutions for very difficult tissues, that may be historical or more recent. We have, of course, asked the referring lab to place all future freshly dissected tissues immediately into 10%NBF. Thank you for any assistance you are able to offer.

Yours sincerely
Alan Taylor BSc(Hons). FRMS
Microtechnical Services
Exeter. UK

Microtechnical Services - for technical excellence in ...<http://www.microtechnicalservices.co.uk/>
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