Venkataramani Sowmya, Davitt Kristina M, Xu Heng, Zhang Jiayi, Song Yoon-Kyu, Connors Barry W, Nurmikko Arto V
Department of Physics, Brown University, Providence, RI 02912, United States.
J Neurosci Methods. 2007 Feb 15;160(1):5-9. doi: 10.1016/j.jneumeth.2006.08.004. Epub 2006 Sep 26.
'Caged' compounds are biological molecules that are rendered inactive by a protecting (cage) group. Photocleaving of chemical bonds associated with the cage species with intense UV light results in the release of the active molecules. This technique, called flash photolysis, allows for real-time study of interacting biological molecules and typically involves the use of high intensity lasers or flash lamps to deliver the UV pulse to the biological specimen [Callaway EM, Katz LC. Photostimulation using caged glutamate reveals functional circuitry in living brain slices. Proc Natl Acad Sci USA 1993;90(16):7661-5; Parpura V, Haydon PG. "Uncaging" using optical fibers to deliver UV light directly to the sample. Croat Med J 1999;40(3):340-5; Denk W. Pulsing mercury arc lamps for uncaging and fast imaging. J Neurosci Methods 1997;72(1):39-42]. Here, we introduce compact, custom-designed semiconductor UV light-emitting diodes (LEDs) as a viable and efficient source for performing flash photolysis studies, focusing specifically on the application of these devices for uncaging neurotransmitters locally onto neurons cultured on artificial substrates. The illumination design feature incorporated in these devices allows for direct placement of the UV source in immediate proximity with the neuron of interest and provides a means for optical triggering of activity in the neuronal culture.
“笼状”化合物是一类生物分子,它们因保护(笼状)基团而处于无活性状态。用强紫外光光解与笼状物质相关的化学键会导致活性分子的释放。这种技术称为闪光光解,可用于实时研究相互作用的生物分子,通常涉及使用高强度激光或闪光灯向生物样本传递紫外脉冲[卡拉韦 EM,卡茨 LC。使用笼状谷氨酸进行光刺激揭示活脑切片中的功能电路。美国国家科学院院刊 1993;90(16):7661 - 5;帕尔普拉 V,海登 PG。使用光纤“解笼”将紫外光直接传递到样本。克罗地亚医学杂志 1999;40(3):340 - 5;登克 W。用于解笼和快速成像的脉冲汞弧灯。神经科学方法杂志 1997;72(1):39 - 42]。在此,我们介绍紧凑的、定制设计的半导体紫外发光二极管(LED)作为进行闪光光解研究的可行且高效的光源,特别关注这些器件在将神经递质局部解笼到在人工基质上培养的神经元上的应用。这些器件中包含的照明设计特点允许将紫外光源直接放置在感兴趣的神经元附近,并提供了一种在神经元培养物中光学触发活动的方法。