Hello,
could anyone please advice on an issue I am facing? We work with precision-cut lung slices (PCLS) and they are often too light/buoyant that when I am embedding them in paraffin (for downstream FFPE histology analyses), the slices often don’t lay flat. This is problematic during sectioning with microtome; when I cut at 4 um, some parts of the tissue are embedded deeper and don’t get sectionined with the rest of the tissue. This often results in tissue sections with a missing part.
Does anyone have a technique they use to ensure their PCLS lay flat during embedding?
thanks!
The PCLS maybe processed with agarose, then washed properly and embedded in agar. After that go for paraffin embedding. Other precautionary measures like use of adhesive slides, proper handling etc are to be maintained. For detailed information, you may login to internet.
Hi Arnold J Apostol ,
I am working on precision-cut kidney slices and facing the same issue. Although kidney slices settle at the bottom of the mold, when the paraffin solidifies, I find that they did not lay completely flat and this results in uneven sectioning. Did you try putting a light-weight mesh on the slice while it is in paraffin?
Arnold J Apostol I did want to share that yesterday after adding molten paraffin on my kidney slice, I tapped the mold on the surface and it helped in laying the slice flatly at the bottom. Maybe this would be useful to you (Provided the lung slice is not so light that it fails to settle.)
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