Microtomy is a technical skill that requires practice and proper training. I suggest you work with a trained histotechnologist to show you the tricks of the trade. Beyond that, make sure that your blade is sharp, that you are not trying to cut too thinly (between 3 and 5 um is the usual thickness), and that your tissue has been properly processed. Note that it's normal for a section to fold a bit when sectioning, this is why you must use a water bath to have it stretch out after sectioning.

Thank you for the quick response! I will attempt to find someone who can assist me further with hands-on training. Also, we're using 6 um. And I am aware of that; however, this is well beyond a little bit of folding.

It helps if the FFPE block is really cold before microtomy. Do you leave the blocks on a cold plate first before attempting to cut?

Brain tissue can take much longer than other tissue to fix in formaldehyde. Check your wax blocks, if there are any soft areas or shrinkage you need a longer fixation period. Slicing the brain to 5mm before fixation can speed this up. Make sure you are using a well tested protocol for the rest of the tissue processing.

If you have no cold plate, a block of ice with a shallow film of water over it will work just as well. The water will help with thermal conductivity and also soften connective tissue.

Trim your blocks in to give a flat surface exposing the full face of the tissue biopsy, then cool them for 10 minutes or more before cutting your sections. Use an untouched part of the blade when any scratches appear in the sections. If the sections are rolling up or not forming a ribbon try thinner sections. 4um should work well. A very gentle brush stroke run over the cutting edge of a new blade can also help ribbons to form.

Good luck!

A good reference book is the "Theory and practice of Histological Techniques".. Lots of editions, originally compiled/edited by Bancroft & Stevens. Many chapters written by ACTUAL practicing Histotechnologists. You can pick up older editions very cheaply on the interweb thingy... Fix your tissue well, process it well and it will section well. Happy new year!

Thank you everyone for your suggestions! To answer the first question: Wei-Qiang Leow, I chill my blocks in a freezer before use; however, I do not take any measures to keep them cold during the sectioning process. Second, Peter Walker, thank you for the suggested text! I will have to check that out.

Hi Svenson!

Few points you can follow:

1. Fix and embed your tissue properly

2. Use new and sharp blade

3. Make your block cold before you start sectioning

4. Moisten the cutting surface of your block with water

5. Practice more to gain a rhythm because proper sectioning is a skill.

Hope this will help. All the best.....

Hello Svenson,

Major problem you cannot get good tissue sections “Surface of your paraffin block is too much dried !!!”

Follow this:

1. Make sure you have enough humidity in the room you keep the microtome. If not, use a humidifier to make the air humid (With no sudden air movements; keep doors always closed when you use the microtome)

2. Pre-cut your tissue block at 10um thickness until you see the tissue expose to surface (make sure you adjust the microtome blade at 45 angle, otherwise the surface will not be equal at all points (this will make tissue slices in different thickness, making your HE stanning or IHC gives false results/observations)

3. Keep the tissue block you pre-cut in -20C freezer, 4C fridge (surface up) or on ice (surface down) for 10-15 min (You will see the surface is cold and can proceed with sectioning)

4. Briefly, do one round with 4um thickness section to remove excess moisture on surface and you will be able to get serial sections without any crumbles afterwards (Make sure you get only transparent sections (This will allow you to see the cells more clearly. You can use a toothpick/small paint brush to avoid sections folding and using that , you can transfer your sections to water bath; make sure you will not keep brain slices in water bath for long ,once you will see the paraffin section surface is clear without folds ,take it ASAP to a silane coated slides and keep in incubator (37C) until use for staining)

I hope this helps!!

Charith