Dear RG community,
Histology related question:
Can anyone advise on how many ffpe sections (estimated) can be cut using the same part of a microtome blade?
The tissue I am sectioning is smooth, unsinewy and contains no bony or calcified structures!
I am having the problem where some sections are peeling or disintegrating on the slides during multiplex staining and I want to ensure this isn’t caused by poor sectioning!
I would rather not have to add in a further NBF step!
Many thanks
Hello Pia Leete,
Formalin-Fixed Paraffin-Embedded (FFPE) tissue specimens are sometimes difficult to cut because the blades must be tissue-specific (some require flexible blades and others, on the contrary, very rigid -without even containing calcified bones or structures-). Make sure you use the right blades.
Also, try to see what the fixing fluid was (some make the tissue hard and "calcify" the tissue during the process).
Moreover, it is sometimes necessary to change the blade (or at least the area of the blade that has been used) after only a few dozen "cuts", corresponding to a few blades if you put several cuts per blade. Again, it all depends on your samples, their fixation and your blades.
Finally, it is sometimes preferable to put the samples at 4 degrees for about 1 hour before cutting them in order to avoid the peeling or disintegrating phenomenon. In some cases, it is even possible to put an ice cube on the fragment to cool it down even more (be careful however to avoid thermal shocks that can fracture the paraffin).
Hoping that this comment will be useful. Feel free to discuss or share some informations about your technical study.
Ludovic Dumont, Ph.D.
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