Multi-overlap immunofluorescence techniques used in nerve cells based on morphology in brain injury models / 中华神经医学杂志

ABSTRACT

Objective To explore the application of multi-overlap immunofluorescence techniques in the morphological research of nerve cells,and provide a new method of targeted research on nerve cells.Methods Ten Balb/c mice were selected and randomly divided into experimental group of severe traumatic brain injury (STBI) and sham control group (n=5);mice in the experimental group were induced STBI models by controlled cortical impact (CCI) method,and the neurological severity scale (NSS) scores were 8 in them.All animals were terminated on day 7.The gene and protein expressions of neuronal nuclei antigen (NeuN),glial fibrillary acidic protein (GFAP) and bromodeoxyuridine (BrdU),and the morphology expression of4',6-diamidino-2-phenylindole (DAPI) were observed under confocal microscopy by applying multi-overlap immunofluorescence techniques.Results The images got by multi-overlap immunofluorescence techniques possessed the characteristics of bright and clear color,strong morphological sense on the expressions.The more obvious expressions of BrdU+NeuN+GFAP+DAPI in the STBI group could be noted as compared with those in the sham control group;the relative expression quantity of BrdU+NeuN+DAPI was significantly different between the two groups (P＜0.05).Conclusion STBI in mice could induce self-regenerated phenomenon on cerebral neural cells or astrocyte;these new techniques provide new scientific means in the targeted study of development,proliferation,apoptosis or death of nerve cells and its mechanism.