Grutzendler Jaime, Tsai Julia, Gan Wen-Biao
Department of Physiology and Neuroscience, New York University School of Medicine, 540 First Avenue, New York, NY 10016, USA.
Methods. 2003 May;30(1):79-85. doi: 10.1016/s1046-2023(03)00009-4.
Particle-mediated ballistic delivery of fluorescent dyes has been recently used to label neuronal populations in a rapid and efficient fashion. Here we describe detailed protocols for this technique as well as recent improvements in its implementation. This technique allows rapid labeling of entire neurons in a Golgi-like manner after membranes of individual neurons are contacted by particles coated with lipophilic dyes. Neurons can be labeled by dyes of different colors at controlled densities to facilitate the study of structural interactions between cells. Furthermore, in conjunction with other histochemical labeling methods, the technique can be used to study changes in neuronal structures associated with pathologic processes in animal models or postmortem human brain. In addition to lipophilic dyes, water-soluble molecules such as calcium indicators can also be delivered efficiently with this technique. The method of ballistic delivery of indicators thus provides new avenues to probe the structure and function of the nervous system.
近年来,粒子介导的荧光染料弹道递送已被用于快速高效地标记神经元群体。在此,我们描述了该技术的详细方案及其实施过程中的最新改进。该技术允许在用亲脂性染料包被的粒子接触单个神经元的膜后,以类似高尔基染色法的方式快速标记整个神经元。可以用不同颜色的染料以可控密度标记神经元,以促进对细胞间结构相互作用的研究。此外,结合其他组织化学标记方法,该技术可用于研究动物模型或死后人类大脑中与病理过程相关的神经元结构变化。除亲脂性染料外,水溶性分子如钙指示剂也可以通过该技术有效递送。因此,指示剂的弹道递送方法为探究神经系统的结构和功能提供了新途径。