小鼠脏器荧光信号的观察

Observation of the fluorescent signals in the organs of mice

目的:观察小鼠肝脏、脾脏和肺内的荧光信号,以及硫酸铜消除自发荧光的效果。方法:(1)细胞实验:将大鼠脑胶质瘤细胞系RG2分成对照组和处理组。细胞成熟贴壁后,4%多聚甲醛固定,处理组用 1.0 mmol/L硫酸铜溶液(经过醋酸铵溶液缓冲)处理10 min,对照组未经硫酸铜处理,直接荧光胶封片,观察2组细胞荧光强度。(2)动物实验:将C57/BL6小鼠分为正常组和尾静脉注射组。尾静脉注射组小鼠经尾静脉注射100 μL RG2.0细胞悬液,3 d后处死,经心脏灌注后取肺、肝脏和脾脏,制作成冰冻切片。正常组小鼠直接经心脏灌注后收集肺、肝脏和脾脏。观察各脏器荧光信号以及硫酸铜溶液处理效果。结果:RG2经硫酸铜处理后荧光信号减弱。小鼠各个脏器的荧光分布不一:肺脏中基本无自发荧光;肝脏无明显自发荧光;脾脏中存在大量自发荧光,且经硫酸铜溶液处理也无法完全消除。尾静脉注射组小鼠在肺和肝脏中可以明显观察到注入的荧光信号。结论:硫酸铜能减弱细胞荧光强度,能够去除脾脏一定程度的荧光背景;肺脏和肝脏可以作为更好的荧光信号观察器官。

Objective： To observe the fluorescent signals in the liver, spleen and lung of mice and the quenching effect of autofluorescence by copper sulfate. Methods： ①Cell experiments： RG2 （rat glioma cell line） cells were divided into the control and treatment groups. Following maturity and attachment, two groups were fixed by 4% PFA. The treatment group was treated with 1.0 mmol/L copper sulfate solution （ammonium acetate buffer） for 10 min while the control group was not treated by the solution mentioned above. After sealed with fluorescent gel, the fluorescent intensity of these two groups was observed. ②Animal experiments： C57/BL6 mice were divided randomly into normal group and tail vein injection group. In tail vein injection group, the mice were injected with 100 μL of RG2 cell suspension and were sacrificed on the fourth day. After cardiac perfusion, the lungs, livers, and spleens were collected for frozen sections. In normal group, after cardiac perfusion, the lungs, livers, and spleens of mice were collected directly. The fluorescent signals of each organ and the effect of copper sulfate solution were observed. Results： After the treatment of copper sulfate, the fluorescent signals of RG2 reduced. The fluorescence distribution in each organ in mice was different; there was almost no autofluorescence in the lung; no obvious autofluorescence in the liver; marked autofluorescence in the spleen, which could not be eliminated by the treatment of copper sulfate solution. In contrast, the green fluorescence signals in the lung and liver in the tail vein injection group could be observed obviously. Conclusion： Copper sulfate can reduce the fluorescent intensity of cells and remove the background signal of spleen to a certain extent; the lung and liver can serve as better organs for the observation of fluorescence signals.