激光拉曼光谱系统无损鉴别大鼠原代胰岛α、β细胞

A LASER RAMAN SPECTROSCOPY SYSTEM FOR NON-DESTRUCTIVE IDENTIFICATION OF RAT PRIMARY PANCREATIC ISLET α AND β CELLS

目的:利用激光拉曼光谱系统无损区分贴壁生长的大鼠原代胰岛α、β细胞,并比较该方法与传统方法的优劣。方法:收集贴壁生长于定位格子培养皿中大鼠胰岛细胞的拉曼光谱、坐标及照片后应用免疫组化结合培养皿坐标定位确定胰岛细胞的分类,用主成分分析结合线性判别分析(PCA-LDA)对两种胰岛细胞拉曼光谱进行分析,用线性判别分析法对两种细胞大小进行分析,两组间光谱及面积差异则采用两独立样本t检验。结果:贴壁生长的胰岛β细胞体积较α细胞大,差异具有统计学意义(P=0.001);然而单纯利用面积胰岛α细胞及胰岛β细胞正确区分率分别为84.5%、50.0%;胰岛α细胞与胰岛β细胞的拉曼光谱有共同特征峰同时也存在显著差异的波峰,即二者有不同的"细胞分子指纹",且将该特点结合PCA-LDA的方法对胰岛α、β细胞进行区分正确率均为100%。结论:激光拉曼光谱系统可以成为快速无损区分胰岛α、β细胞的有效方法。

Objective：To discover a novel non-destructive method using a laser Raman spectroscopy system used for the identification of rat original pancreaticαandβcells,and compare its advantages and disadvantages with the conventional method in purifying pancreatic islet cells.Methods：Rat primary pancreatic islet adherent cells in culture dish were collected for the cell type identification by Raman spectra,coordinates and photos,and assessed by immunohistochemistry in combination with coordinates and photos.PCA-LDA was used to analyze the Raman spectra of the two cell types,LDA was used to analyze their sizes,and ttest was used to compare the differences of their spectra and areas of two type cells.Results：Pancreaticβcells are larger thanαcells（P〈 0.05）.However,when considering cell surface area alone,the accuracy rate for discriminatingαcells is 84.5%,and that forβcells is 50.0%.,Raman spectrum waves in isletαandβcells share certain similarities,and also,some significant differences.Each cell type can be considered to have an individual＂molecular fingerprint＂.Therefore,combination of Raman spectroscopy with PCALDA,we could increase the accuracy rate to 100%in the identification of isletαandβcells.Conclusion：Laser Raman spectroscopy may be a new promising method for the purification of islet cells because of its accuracy and non-invasiveness.