体视学与激光共聚焦显微镜图像分析黑色素瘤细胞三维形态学参数的变化  

Stereological and Laser Scanning Confocal Microscopic Analysis of 3-Dimensional Morphology of Melanoma Cells

作　　者：张莹[1] 胡纳[1] 汪曣[1] 杨春梅[1] 杨立 胡新华 冯远明[1,4] 

机构地区：[1]天津大学精密仪器与光电子工程学院天津市生物医学检测技术与仪器重点实验室 [2]Department of Internal Medicine,Brody School of Medicine,East Carolina University,Greenville,NC 27858,USA [3]Department of Physics,East Carolina University,Greenville,NC 27858,USA [4]天津医科大学肿瘤医院放射治疗科

出　　处：《光学学报》2012年第9期198-205,共8页Acta Optica Sinica

基　　金：国家自然科学基金(81041107;81171342)资助课题

摘　　要：体视学是由形态学与数学交叉形成的一门新兴学科,通过二维结构信息定量测量和分析三维形态结构特征。采用体视学点计数方法,对肿瘤细胞B16F10、B16/Vector及B16/GPR4各20组的共聚焦显微镜图像进行测量,定量分析了其三维形态参数的差异。对比三类细胞的体视学参数发现,对照组B16/Vector和原始细胞B16F10没有明显差异,实验组B16/GPR4的细胞比表面及相对形状因子参数与另两类细胞相比均表现出了显著性差异(p值分别小于0.05和0.02,即B16/GPR4细胞形态光滑,没有较多的突触和伪足),与前期Transwell小室细胞侵袭及迁移实验研究结果相吻合,体现出B16/GPR4细胞中G蛋白耦联受体4(GPR4)基因对细胞迁移性抑制的生物特性。研究结果表明,体视学和激光扫描共聚焦显微镜细胞图像的结合,可以实现细胞三维结构参数的快速测量,是细胞生物学尤其是细胞形态学分析的有效研究方法。Stereology is an interdisciplinary method for 3D morphological study developed from mathematics and morphology. And it is widely used in medical image analysis and cell-biology studies. The laser scanning confocal microscopic images （LSCMI） of tumor cells are quantitatively analyzed by using stereological point counting method. Three groups of cells, B16F10, B16/Vector and B16/GPR4 cells suspended in culture media, are studied and 20 image sets of each are taken and used in the experiments. The results show that the control group B16/Vector has no significant differences as comnared with the parental B16F10 cells, while the experimental group B16/GPR4 shows significant differences in the stereological parameters of the cellular surface area to volume ratio （Rsvc） and the relative cellular form factor （f） （io^0.05 for R and p%0.02 for f, respectively）, suggesting that B16/GPR4 cells have less surface protrusions and reduced irregularity. This is in line with the previous observations that GPR4 inhibits migration and invasion of tumor cell in the transwell assay. The combination of stereology and laser scanning confocal microscopic imaging techniques provides a quick and useful method in cell biology, and especially in cell morphology studies.

关 键 词：医用光学 肿瘤细胞三维形态 体视学 激光扫描共聚焦显微镜 B16F10黑色素瘤细胞 G蛋白耦联受体4 

分 类 号：R318.51[医药卫生—生物医学工程]