In the lab, this week, we were working with tissues from chicken with (possible) tuberculosis (the samples came ready for processing from another department of the Faculty). But we had some problems sectioning and staining them.
We did our common protocol for paraffin inclusion¹ (since we are starting to work with pathologic samples), and the tissue was still hard as it damaged the microtome blade. We stained the tissues that were possible to cut, but many fell off. Those that stayed, looks really bad, since they are scratched.
Does anyone has any recommendation for processing and sectioning granulomas?
And what about the missing sections during staining?
Thanks!
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¹Protocol: Alcohol 70° 1h; Alcohol 80° 1 h; Alcohol 96° 1 h; Absolute Alcohol 1 h x3; Xileno 1 h x3; Paraffin 1 h x2 (the samples stay in the last paraffin bath some additional hours, because the program finishes at middle of the night).
Dear colleague
if the mass of granuloma is hard due to calcium salt deposition, you should de-calcified if with formic acid or EDTA, the protocol can early found by simple search in google, but you should note that the sample of granuloma should not rinsed in de-calcification solutions for long time (half the time need to de-calcified bone).
Thanks, Saevan.
The person who brought the samples will try decalcifying them and bring us new pieces; although, they said that is not common calcification in chicken granulomas.
We will probably try to do a reinclusion of the blocks with large time in a paraffin bath, as a technician recommend to us.
Please share your experience if problem solved with above suggestions or not
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