If I understood you well you have a 11 hours protocol of which 10 hours are fixation? If this is true I think your processing steps (other than fixation) are too fast even if rat tissues are leaner than human.
If in spite of 10 hours fixation you still have reeddish centers in the liver slices: how thick they are? How many they are per cassette?
You should "open" the liver samples (the ones going into the first fixative after necropsy) to assure a good fixation. I think you should prepare the cassettes immediately after necropsy and set the slices to fix.
If you cassette after fixation and place the cassettes in the tissue processor you could embed the next and section the morning which will reduce the TAT.
I did not understand the "drying overnight" step you describe. Why so long?
I think you have a mixed problem between incomplete initial fixation and too fast tissue processing (1 of 11 hours protocol).
Jamie E Erickson wrote:
I hope someone can help with this problem. I am working on a
toxicology study that requires sections of Heart, liver and Lung from rats
on study . These studies we want a quick turn around so we get the tissues
from necropsy in the afternoon, livers are scored at necropsy and put
into formalin jars. I gross the samples Liver (left, midial, caudal lobes
into one cassette) the next morning done by 11am (liver is still pink in
the middle). The samples are put on the processor (11 hour processor run)
that night with formalin in the first station so fixation after grossing
is 10 hours. The next day I embed and section the samples and dry the
sections overnight. Slides are stained and given to the pathologist that
My problem is the liver samples all other blocks are fine, the liver
blocks section fine but on drying either in a rotating drying oven
vertically or on the bench at room temp overnight some NOT all of the
liver samples have lots of cracks and some fall off. It very much looks
like areas of water as it dried causes these cracks. Anyone have ideas to
remedy this problem..?? They want to keep this turn around time if
possible and liver is a key read out... Any ideas...
Sr. Research Associate
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
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