SEMIAUTOMATED ANALYSIS OF NMDA-MEDIATED TOXICITY IN DIGITIZED COLOR IMAGES FROM RAT HIPPOCAMPUS

The evaluation of neuronal cell survival after, for example, mechanica l, hypoxic or drug-mediated injury requires the analysis of a high num ber of histological specimens. Since this is a time-consuming occupati on, we have developed a semi-automated analysis routine for the determ ination of the distribution of live and dead cells. After digitalizati on of the histological preparations, 8-bit colour bitmaps were assesse d using a compiled image-analysis programme of the software package Kh oros. In the current study a detailed example of the application of th is image-processing approach is described for the investigation of the cell survival after intraventricular application of N-methyl-D-aspart ate (NMDA). The samples were prepared as fuchsin acid/toluidine blue s tained hippocampal thin slices. The calculated areas of the live and d ead cells were highly correlated with manual counts of live and dead c ells in the 100 samples examined in this study. Twenty-four hours foll owing NMDA-treatment animals (n = 5) were found to have significantly fewer live and more dead hippocampal cells than the saline-treated ani mals (n = 5), using either automated or manual examination techniques. The automated technique also revealed that NMDA treatment resulted in a reduction in the density of live cell distribution. (C) 1998 Elsevi er Science B.V. All rights reserved.