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The thickness of section was 15 20 μm.
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When using the freezer microtome, the thickness of sections was set to 20-40 μm.
The shrinkage along the z-axis was estimated by comparing the thickness of sections as they come off the microtome and after histological processing with Neurolucida.
The thickness of sections produced using paraffin-embedded specimens did not allow accurate co-localization of CD3+ and CCR8+ cells.
This latter value is below the limits of experimental error in histological assays, which may arise from different local thickness of sections or uneven distribution of the antibodies.
We then applied the Abercrombie formula (Abercrombie 1946), which uses the average measured diameter of the germ cell nuclei and the thickness of sections to correct for any double counting due to single cells appearing in two successive sections.
Even their histological demonstration is dependent on the commitment of the pathologist[ 17 ],[ 18 ]; the number of lymph nodes examined, the number and thickness of sections taken and the use of immunhistochemistry and molecular analysis[ 14, 15, 18 22 ].
Thus, estimation of nuclear DNA values on sections may be sufficiently precise only if the thickness of each individual section is known and the nuclear IOD-values have mathematically been corrected for the error caused by different thickness of sections [ 33].
We then used the Abercrombie formula (Abercrombie 1946), which uses the average measured diameter of the germ cell nuclei (D) and the thickness of sections (S) to correct for any double counting due to single cells appearing in two successive sections, to obtain the true count (TC): TC = CC × S/(S+ D).
The thickness of sections also affects the style of the braid.
Total volume of each section was calculated (area of the section x thickness of the section) and follicle counts for each animal were corrected for the total volume of ovarian tissue counted.
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