Abstract:

In order to identify the influence of three fixation solutions on the quality of paraffin section of turbot liver tissue, and select the best fixation solution, in this study, three fixation solutions, Bouin fixation solution, Davidson fixation solution and 4% Paraformaldehyde (PFA), were used to fix the turbot liver tissue, and paraffin sections were prepared. The morphological characteristics of the tissue cells were observed by hematoxylin-eosin (H-E) and immunohistochemical staining. The integrity and immunohistochemical positive rate of hepatocyte were analyzed by statistical methods. The results showed that there were significant differences in the fixation effect of the three fixation solutions on paraffin sections of turbot liver tissue. Clear structure and complete morphology of hepatocyte were observed after H-E staining with Bouin fixation solution. However, the hepatocyte manifested dissolved and unclear structure by Davidson fixation solution, but the nucleocytoplasmic boundary could be distinguished. 4% PFA fixation solution showed bright color of hepatocyte, but the boundary of nucleus and plasma was not clear. The percentage of intact hepatocytes after 4% PFA fixation was significantly lower than that after Bouin and Davidson fixation solutions treatment (P<0.05), Bouin and Davidson fixation solutions showed no significant difference (P>0.05). There were significant differences in the positive rate of hepatic proliefrating cell nuclear antigen (PCNA) immunohistochemical staining among the three fixation solutions, and that of Bouin fixation solution was significantly higher than 4% PFA and Davidson fixation solutions (P<0.05). In conclusion, Bouin fixation solution is the optimal selection for turbot hepatic H-E or immunohistochemical staining for observing tissue cell structure and cell proliferation.

Key words: fixation solution, hematoxylin eosin staining, immunohistochemistry, PCNA, liver