Image analysis of liver cell nuclei from sodium valproate-treated developing rats

Liver imprints were obtained from male Wistar rats that after a 21-day age received 100 mg/kg VPA daily for 9, 14 and 39 days. The preparations were fixed in an absolute ethanol-acetic acid mixture (3:1, v/v) for 5 min and subjected to the Feulgen reaction for studying nuclear features evaluated with image analysis using KS400-version 3 software.

The nuclear features obtained were as follows: total area (µm2), absorbing area (µm2), perimeter (µm), Feret ratio, optical density (= absorbance), integrated optical density, variability in the degree of chromatin packaging (SDtd), nuclear entropy, and nuclear energy. Animal care and use protocols were approved by the Ethics Committee for Animal Use at the University of Campinas (registration no. 6269-1/2023) and followed the guidelines established by the Brazilian National Council for the Control of Animal Experimentation (CONCEA).