利用双光子显微镜检测活体动物脑内Ca^(2+)动态变化  

In vivo Imaging of Ca^(2+) Signaling Using Two-photon Laser Scanning Fluorescent Microscopy

作　　者：刘双双[1] 廖美华[2] 尹伟[1] 林赵肖楠 肖桂凤[1] 

机构地区：[1]浙江大学医学院公共技术成像平台,浙江杭州310058 [2]浙江大学毒理与生化药学研究所,浙江杭州310058

出　　处：《激光生物学报》2014年第1期29-32,24,共5页Acta Laser Biology Sinica

基　　金：浙江省科技厅分析测试项目(2012F82G2010024);浙江大学实验技术研究项目(20130626)

摘　　要：利用正置双光子显微镜系统和荧光探针标记技术,观察脑内Ca2+分布,建立测量活体动物脑内Ca2+动态变化的实验方法。制作活体动物颅骨开窗样本,脑内负载Ca2+标记物Oregon Green 488 BAPTA-1和星型胶质细胞标记物Sulforhodamine 101,利用双光子显微镜分别检测神经元和星型胶质细胞内Ca2+分布和动作电位引起的Ca2+瞬变。结果显示双光子显微镜可探测到脑内250μm处荧光信号,图像清晰且信噪比高,并能实时检测神经元和星型胶质细胞内Ca2+信号的动态变化。活体脑内Ca2+检测技术平台的建立为基础研究和医药应用提供了在体实验依据。Using two-photon laser scanning microscopy and fluorescent probe labeling technique,we established a new method for observing the distribution and variation of Ca2＋signaling in the mouse brain in vivo.Cranial window surgery of anesthesia mouse was made,Ca2＋marker Oregon Green 488 BAPTA-1 and astrocyte marker Sulforhodamine 101 were loaded into the brain,and then Ca2＋distribution and Ca2＋transients were detected via two-photon microscopy.The re-sults showed that fluorescent signals were able to be detected clearly in the brain at a depth of up to 250 μm with high signal to noise ratio.Ca2＋transients was observed in both neurons and as astrocytes.It indicates that the platform for de-tecting Ca2＋signaling in vivo by two-photon microscopy has been successfully established,and this platform may provide valuable information for basic research and medical applications.

关 键 词：双光子显微镜 活体动物 Ca2+成像 神经元 星型胶质细胞 

分 类 号：Q-334[生物学]