Page 140 - Contributed Paper Session (CPS) - Volume 3

CPS1972 Livio Corain et al.
months old), were obtained from local abattoirs in the Veneto region. Animals
were treated according to the present European Community Council directive
concerning animal welfare during the commercial slaughtering process and
were constantly monitored under mandatory official veterinary medical care.
The posterior domain (lobules VIII and IX) of each specimen was cut into 8
μm thick parasagittal sections. For each cerebellar sample, one section every
five was stained (a total of 10 slides per individual per sex) according to the
Nissl protocol (Cozzi et al., 2017). Ten stained sections per subject per sex were
scanned with a semi-automated microscope equipment at a magnification of
40x in fast mode with automatic focusing, saving the acquisition as Jpeg2000
images. The topographical organization of the bovine cerebellum was
analyzed on the Nissl-stained coronal sections, and the histology of the
cerebellar cortex resulted uniform over the entire lobules VIII-IX of the vermis
(Figure 1.A,1.B,1.C). Three layers were distinguishable from the outer to the
inner cerebellar cortex: (i) the superficial molecular layer, (ii) the Purkinje layer
and (iii) the inner granular layer (Figure 1.D,1.E,1.F).
The complete analysis of the acquired imaged of cerebellar slices involves
the detection and outline of tens of thousands of cells. This is not feasible by
human annotation of the images, unless the procedure is carried out in small
region of interest, potentially introducing bias in the procedure. To tackle the
problem, we developed an automatic procedure (Grisan et al., 2018) that can
process the images identifying the position and outline of most of the visible
cells, taking care of the different sizes among the different cell populations,
and at the same time addressing the packed and clustered appearance of cells
in the different layers of the cerebellum, and in particular in the granule cells
layer, where they are most packed.

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