摘要
目的:通过下丘脑弓状核乙酰胆碱表达的检测、图像处理、图像分析,应用阳性神经元面积百分比(APPN)和染色灰度相对强度值(RISGL)相结合的方法,探讨免疫组化(IHC)图像定量分析的特点。方法:样本来源于运动性免疫抑制过程中SD大鼠下丘脑弓状核胆碱能阳性神经元乙酰胆碱表达的免疫组化切片,包括第0、2、4、6周与每周Control、Immediately after exercise、3 hours after exercise三组共计12组(n=6)。采用免疫组化技术测定其乙酰胆碱(ACh)的表达,并根据ACh阳性神经元总面积(TAPN)、阳性神经元灰度平均强度值(AISGL)、APPN、RISGL、APPN/RISGL参数进行免疫组化图像的定量分析,从而比较APPN和RISGL在定量分析中与传统参数的不同及其优势。结果:APPN与TAPN的变化呈现几乎一致的变化特征,能发现相应差异的显著性意义(P<0.05),但APPN的敏感性和抗干扰性更强。RISGL与AISGL呈现的结果并不完全一致,而且APPN结合RISGL比单参数更能反映阳性表达情况。结论:APPN和RISGL免疫组化图像分析参数能较可靠地、准确地进行图像定量分析,APPN与RISGL的结合不仅可反映阳性表达的数量情况,还可以帮助分析其作用机理,优于传统分析参数。
Objective： The acetylcholine expression in hypothalamus arcuate nucleus is detected and then the images are processed and analyzed. The features of the image quantitative analysis of immunohistochemistry （IHC） with the method combining two parameters of area percent of positive neuron （APPN） and relative intensity of staining grey level （RISGL） were investigated. Methods： Samples were the im- munohistochemical slices of acetylcholine（ACh）expression of hypothalanlic arcuate nucleus cholinergic neurons in the process of exercise induced immunosuppression, which included twelve groups of ＂0 w, 2 w, 4 w, 6 w＂ and three groups of ＂control, immeditative after exercise, 3 hours after exercise＂ in every week. IHC technology was used to detect the ACh expression. The image quantitative analysis of IHC was conducted in accordance with the parameters of ACh total area of positive neuron （TAPN）, average intensity of staining grey level （AISGL）, APPN, RISGL, APPN/RISGL. Then the differences among APPN, RISGL and traditional parameters in the quantitative analysis were com- pared and the advantages were found. Results： The changes of TAPN and APPN showed almost the same variation. Namely the corresponding significant differences could be found through these two parameters（ P 〈 0.05）, but the sensitivity and anti-interference of APPN was higher. The results of AISGL and RISGL were not coincident completely. Furthermore, with the combination of APPN and RISGL, the positive expression could be reflected better than any single parameter. Conclusion： The parameters of immunohistochemical image analysis, APPN and RISGL, can be reliable and accurate in image quantitative analysis of IHC. The combination of APPN and RISGL can not only reflect the expression of positive neurons, but also help analyze its mechanism, which is better than traditional analysis parameters.
作者
曾志刚
邱红
朱梅菊
朱洪竹
萧建华
ZENG Zhi-gang QIU Hong ZHU Mei-ju ZHU Hong-zhu XIAO Jian-hua(School of Physical Education Department of Pediatrics of the Mfilialed Hospital, Jinggangshan University, Jiangangshan 343009, China)
出处
《中国应用生理学杂志》
CAS
CSCD
2017年第3期282-286,共5页
Chinese Journal of Applied Physiology
基金
国家自然科学基金资助项目(31160217
31360255)
江西省教育厅科技项目(GJJ10545)
关键词
免疫组化
图像分析
面积
灰度
大鼠
immunohistochemistry
computer-assisted image processing
small-area analysis
grey level
rat
