Simultaneous multi-parameter observation of Harring-tonine-treating HL-60 cells with both two-photon and confocal laser scanning microscopy

Harringtonine (HT), a kind of anticancer drug isolated from Chinese herb-Cephalotaxus hainanensis Li, can induce apoptosis in promyelocytic leukemia HL-60 cells. With both two-photon laser scanning microscopy and confocal laser scanning microscopy in combination with the fluores-cent probe Hoechst 33342, tetramethyrhodamine ethyl ester (TMRE) and Fluo 3-AM, we simulta-neously observed HT-induced changes in nuclear morphology, mitochondrial membrane potential and intracellular calcium concentration ([Ca2+]i) in HL-60 cells, and developed a real-time, sensitive and invasive method for simultaneous multi-parameter observation of drug- treating living cells at the level of single cell.

基于双螺旋点扩展函数工程的三维多焦点双光子激光扫描显微技术

为了进一步提高成像速度和分辨率,提出了基于双螺旋点扩展函数(DH-PSF)工程的多焦点双光子激光扫描显微成像方法和系统(DH-MTPLSM)。在激发光路中,通过高速相位型空间光调制器(SLM)同时实现了三维多焦点阵列的产生和在样品面上的高精度并行数字寻址扫描;在探测光路中,通过双螺旋相位片将系统探测PSF调制为DH-PSF,从而提供样品的轴向信息,减少轴向扫描层数,进而提高三维成像速度;结合基于DH-PSF的数字重聚焦算法,恢复出不同深度样本的宽场图像,通过单次二维扫描获得样品的三维光切片信息。在此基础上,利用搭建的DH-MTPLSM系统开展了小鼠肾组织切片的双光子成像实验,验证了该方法的快速三维高分辨成像能力,这对于MTPLSM的发展具有重要的意义。