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Author Topic: Thick paraffin sections 40 mikrom  (Read 3085 times)

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Offline voland

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Thick paraffin sections 40 mikrom
« on: September 28, 2009, 02:22:00 AM »
Hi,

I need some help with cutting thick parrafin 40mikrom rat brain sections for stereology. I cut them with a microtome but they roll up when putting them in the water bath. Anyone knows how to solve this problem?

Thanks

Thick paraffin sections 40 mikrom
« on: September 28, 2009, 02:22:00 AM »

Offline richard03

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Re: Thick paraffin sections 40 mikrom
« Reply #1 on: September 28, 2009, 11:26:48 PM »
The best way to cut 40um rat brain sections is to cut it using warm water.

Step 1. Immerse a brush in water bath with temperature usually around 40-45 C.
Step 2. Warm the paraffin block surface with the brush back and forth several times so the block surface is wet and warm.
Step 3. Cut slowly while brush is still gently toughing the block surface and moving brush along with sections together. Now you got one single section.
Step 4. Pick up the section gently with your finger and put it in water bath from one end first and then gently move section down to the other end.
Step 5. Wait for it to be flatten in a few seconds or minutes depending on water bath temperature, and then mount on charged or coated slides.
Step 6. Allow the slides air dry overnight.
Step 7. Put the slides in 45 C oven (never go over 50 C!!!) and bake overnight.
Step 8. Proceed with normal staining procedure.

Reference:

http://www.ihcworld.com/_protocols/histology/gelatin_slides.htm

Re: Thick paraffin sections 40 mikrom
« Reply #1 on: September 28, 2009, 11:26:48 PM »