Abstract
An image analyzing system (Luzex 500, Japan Regulator Ltd., Hachioji, Tokyo) was tested for its usefulness by examining red blood cells of guinea pigs, mouse and guinea pig macrophages and L2C cells of leukemic strain 2 guinea pigs for their maximum and Feret's diameters and areas. Effects of muramyl dipeptide (MDP) on guinea pig macrophages were also investigated by determining these parameters on the cultured macrophages in the presence of 10 μg/ml of MDP in a medium supplemented with fetal calf serum. Several factors which affected the analysis were also examined. From a total of 1, 158 red blood cells of a guinea pig, 7.3±0.8μm, 8.2±0.8μm and 39.8±6.6μm^2 were obtained for their Feret's and maximum diameters and areas, respectively. These measurements were not affected by differences in the threshold level provided that an optimal threshold level was set for a particular field under the microscope. Mouse macrophages were shown to be heterogenous in their sizes as revealed in a similar examination on a cultured preparation. Their measurements were 21.5±7.9μm, 17.9±7.3μm and 137.9±41.2μm^2 for maximum and Feret's diameters and areas, respectively. Guinea pig macrophages were also heterogenous in their sizes and their maximum and Feret's diameters and areas were 18.5±2.3μm, 16.8±2.7μm and 226.6±61.3μm^2, respectively. L2C cells taken from a leukemic guinea pig showed 9.6±1.7μm, 11.0±1.8μm and 70.9±21.6μm^2 for those dimensions, which were consistent with the previous data reported by others but much larger when examined on a cytocentifuged and stained perparation of cells which had been separated on Ficoll-Hypaque gradients. Their dimensions were 18.8±2.0μm, 17.3±2.1μm and 223.9±41.4μm^2 for those parameters, respectively. Guinea pig macrophages which were cultured in the presence of 10 μg/ml of MDP were slightly larger in these dimensions than those in the control experiment which did not contain MDP, their measurements being 29.8±11.5μm, 24.6±10.9μm and 291.8±98.0μm^2 for the former and 36.3±12.7μm, 27.2±11.2μm and 304.1±91.2μm^2 for the latter, respectively. These results show that the image analyzing system is potentially useful for the morphological studies under various conditions involving a static status as well as dynamic changes of many types of cells.
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