D. Manahanvaughan et al., SEMIAUTOMATED ANALYSIS OF NMDA-MEDIATED TOXICITY IN DIGITIZED COLOR IMAGES FROM RAT HIPPOCAMPUS, Journal of neuroscience methods, 82(1), 1998, pp. 85-95
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.