[Histonet] paraffin sectioning problems

From:James Dooley

I recently started paraffin sectioning and I am having trouble with getting good quality sections reproducibly. I was wondering if you have any tips that may help. I am sectioning thymus, pancreas, and lung. Section thickness is 10 um. The microtome that I am using is Reichert-Jung Supercut, disposable knife holder by Leica, and Accu-Edge low profile blades. I fix the tissue in a modified Carnoy's fix overnight, dehydrate in graded alcohol, 100% ETOH, Toluene 2x15 minutes, Paraffin at 65 degrees overnight with 2X changes of paraffin. Embed and store at RT. The paraffin I am using is Fisherbrand Paraplast X-tra Tissue embedding medium. Any suggestion would be greatly appreciated. 

Thank you in advance,
James Dooley

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