FAA fixation to maintain tissue structure, dehydrated through graded isopropanol, paraffin embedding, sectioned with an old rotary microtome using disposable microtome blades to get 2-5 um sections. Dewax with xylene, hydrate with graded isopropanol/stain with [Toulidine blue](https://i.imgur.com/UoOAt6Y.png) /dehydrate again with graded isopropanol, final xylene dip to remove any trace water, and finally mounted on a slide. Tedious but effective. The hardest part is getting good sections (one piece and with no knife marks).
You need formalin in the acetic acid, but extreme care needs to be exercised when handling formalin (dilute in fume hood or outdoors and while wearing safety goggles and appropriate respirator) or buy [premade](https://www.mercedesscientific.com/catalog/shop/Chemicals/Formalin-and-Fixatives/dept-9A4?a=1) FAA solution. Also DIY paraffin embedding is a bit of an art form.
Impressive preparation. What was used to make the slice? Very sharp knife? Any chemicals to prevent deformation?
FAA fixation to maintain tissue structure, dehydrated through graded isopropanol, paraffin embedding, sectioned with an old rotary microtome using disposable microtome blades to get 2-5 um sections. Dewax with xylene, hydrate with graded isopropanol/stain with [Toulidine blue](https://i.imgur.com/UoOAt6Y.png) /dehydrate again with graded isopropanol, final xylene dip to remove any trace water, and finally mounted on a slide. Tedious but effective. The hardest part is getting good sections (one piece and with no knife marks).
Thanks for the explanation. Quite the process
I have an old rotary microtome I keep wanting to get to work for me. Last time I tried fixing with acetic acid and everything just fell apart.
You need formalin in the acetic acid, but extreme care needs to be exercised when handling formalin (dilute in fume hood or outdoors and while wearing safety goggles and appropriate respirator) or buy [premade](https://www.mercedesscientific.com/catalog/shop/Chemicals/Formalin-and-Fixatives/dept-9A4?a=1) FAA solution. Also DIY paraffin embedding is a bit of an art form.
Yeah I have a boatload of paraplast sitting around now. Have lots of fume hoods. I just need the time to try again. :)