A. Two phases of measurement procedure
To reduce the time of interaction with the operator and to increase the effectiveness of his participation in the measurement procedure, we divided the planning of the scanning procedure and measurement itself in two separate parts. The cortical plate represents on the section the wide strip of cellular elements specifically arranged in the space. The geometry of this organization is very complex. So far, the scanning procedure has to follow the geometry of the cortex and the orientation of neuronal columns.
The step of the measurements, called as “Scanning layout creation”, was interactive. The researcher could define the limits and the orientation of the measured fragments of each section under the low microscopic magnification to orient the scanning along the cortical layers in one direction and along the neuronal columns in the other.
Generally, the scanning meander was oblique-angled and presented a series of parallelograms (see Fig. 3).
The first part of the program provided to the second part the capability of the automatic scanning of the several serial sections, collected on the slide (Fig.4). During the second step, the fully automatic measurement, the program used the file of coordinates and angles of orientation of the measuring fields, already created during the first step. So far, the neuronal columns were consistently oriented vertically in the measuring mask regardless the actual orientation on the slide (see direction of the columns of blue arrows on Fig. 3). To minimize interactions between the program and the operator during the measurements, the gray-tone coding of the measurement results was provided. The image, which visualized the measured fragments of each section, was stored in gray-tone image memory. After the measurement this picture provided the information for the checking of the result and for data editing.
It seems that today the same approach can easily be extended to the measurements of the coronal sections of the whole brain.