Evaluation of hippocampus in rhesus monkeys after chronic (1-year) inhalation exposure to marijuana: II. Morphometric methods
An innovative method was used to collect data for morphometric analysis of synaptic measurements. Negatives of the CA3 region of the hippocampus were obtained as described by Andrews et al., 1990, and evaluated by computersupported image analysis. Negatives of 4000X and 40,000X magnification were used to measure synaptic density and width, respectively. The image analysis system (IAS) used an IBM-AT compatible computer with a frame grabber (Imaging Research Inc.). This particular system can measure either straight line distance (rubber line) or curved two point distances (draw line).For synaptic density, the IAS was calibrated in angstroms at a magnification of 4000X. A log file was created for each negative in which the measurement for each synapse was stored. The negative was placed on a transparency with grids 9 mm square to ensure that the entire negative was scanned. The image was displayed on a multicsync monitor using a high resolution camera (Applied Intelligent Systems Inc.). When a synapse came into view, the image was digitized on the monitor (Fig. 1 A and B) and stored in digital form.