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.
出处
《激光生物学报》
CAS
CSCD
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)
