Kovalchuk Yury, Garaschuk Olga
Cold Spring Harb Protoc. 2012 Jul 1;2012(7):778-85. doi: 10.1101/pdb.prot070037.
This protocol describes a technique for high-resolution chloride imaging of living cells using a quinoline-based chloride (Cl(-)) indicator dye, MQAE (N-[6-methoxyquinolyl] acetoethyl ester). Bath-applied to acute brain slices, MQAE provides high-quality labeling of neuronal cells and their processes. In living anesthetized mice, cortical cells are labeled using the multicell bolus loading procedure. In combination with two-photon microscopy, this procedure enables in vivo visualization of cell bodies of neurons and astrocytes as well as some astrocytic processes and allows one to monitor changes in the intracellular chloride concentration in dozens of individual cells.
本方案描述了一种使用基于喹啉的氯化物(Cl(-))指示剂染料MQAE(N-[6-甲氧基喹啉基]乙酰乙酯)对活细胞进行高分辨率氯化物成像的技术。将MQAE浴应用于急性脑片时,可对神经元细胞及其突起进行高质量标记。在活体麻醉小鼠中,使用多细胞团注加载程序对皮质细胞进行标记。结合双光子显微镜,该程序能够在体内可视化神经元和星形胶质细胞的细胞体以及一些星形胶质细胞的突起,并允许监测数十个单个细胞内氯化物浓度的变化。