An Improved Protocol for Fluoro-Jade C Staining Method

• 1、College of life science,Hebei Normal Uiversity,Shijiazhuang,050024

Abstract：Fluoro-Jade C stain is a frequently-used method for labeling degenerating neurons in tissue. In the experimental process, we found that the tissue sections were easily broken and dropped from slide glass in frozen section stained with traditional Fluoro-Jade C method. In order to solve this problem, we modified the protocol of Fluoro-Jade C stain, and have achieved satisfactory results. Twelve adult C57BL/6 male mice were randomly divided into control and PD model groups. The PD model was duplicated with intraperitoneal injection of MPTP (30 mg/kg/d) for 5 days. On the eighth day, the mice were perfused and the midbrain were removed and sectioned with freezing microtome. The sections in same position of each sample were stained by Flurometric TUNEL method; traditional Fluoro-Jade C histofluorescence staining and the improved method of Fluoro-Jade C staining to detect the degenerative dopaminergic neurons. The results showed that there was no degenerative positive neurons in SNc in the control group, while there were some positive neurons FJC and TUNEL-labeled neurons in MPTP-treated mouse, and the traditional and improved FJC staining were both labeled the degenerative neurons. Although the improved FJC staining had the higher background, it can clearly show the degenerative neurons. There were no significant difference among the three methods. The modified FJC histofluorescence staining reduced the damage to the slides, and easier combined with other immunofluorescence. So, the modified FJC staining method can be very simple and rapidly detect the degenerative neurons which made the slides without damage.

keywords： Fluoro-Jade C MPTP dopaminergic neurons TUNEL

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