Microfire ccd camera

Design-based stereologic analyses were performed using a stereology workstation based on a Zeiss Axioplan II microscope (Carl Zeiss MicroImaging, Thornwood, NY, USA) equipped with Plan-Neofluar 2.5Â (N.A. ¼ 0.075) and 40Â (N.A. ¼ 1.30) objectives, Fluar 10Â (N.A. ¼ 0.5) and 20Â (N.A. ¼ 0.75) objectives, a Microfire CCD camera (Optronics, Goleta, CA, USA), a motorized stage (Ludl Electronics, Hawthorne, NY, USA), and stereology software (Ster-eoInvestigator, Version 10, MBF Bioscience, Williston, VT, USA).
Regions of interest included the striatum and thalamic centromedian/parafascicular complex. Delineations of these regions were performed according to established criteria [7, 9, 18, 19] . Volumes were calculated using Cavalieri's principle, by measuring the projection area on all sections and multiplying this by the interval of the selected sections, and by the thickness of the sections [20] . The total cell number was evaluated using the optical fractionator probe [21] . The counting frame was 70 Â 70 mm for neurons and 15 Â 15 mm for glial cells. The dissector height was 25 mm and the guard zone was 10 mm. All cells whose nuclear boundaries or (if present) nucleoli came into focus across the dissector height were counted. Finally, the cell density was calculated by dividing the total cell number by the volume of the region.