Georgia Institute of Technology, Coulter Department of Biomedical Engineering, 313 Ferst Drive, Room 2127, Atlanta, Georgia 30332, United States; Emory University, Department of Neurosurgery, 101 Woodruff Circle, WMRB Rm 6337, Atlanta, Georgia 30322, United States.
Emory University , Department of Neurosurgery, 101 Woodruff Circle, WMRB Rm 6337, Atlanta, Georgia 30322, United States.
Neurophotonics. 2016 Apr;3(2):025001. doi: 10.1117/1.NPh.3.2.025001. Epub 2016 Apr 5.
The use of bioluminescent reporters in neuroscience research continues to grow at a rapid pace as their applications and unique advantages over conventional fluorescent reporters become more appreciated. Here, we describe practical methods and principles for detecting and imaging bioluminescence from live cells and animals. We systematically tested various components of our conventional fluorescence microscope to optimize it for long-term bioluminescence imaging. High-resolution bioluminescence images from live neurons were obtained with our microscope setup, which could be continuously captured for several hours with no signs of phototoxicity. Bioluminescence from the mouse brain was also imaged noninvasively through the intact skull with a conventional luminescence imager. These methods demonstrate how bioluminescence can be routinely detected and measured from live cells and animals in a cost-effective way with common reagents and equipment.
生物发光报告基因在神经科学研究中的应用正在迅速发展,因为它们的应用和相对于传统荧光报告基因的独特优势越来越受到重视。在这里,我们描述了从活细胞和动物中检测和成像生物发光的实用方法和原理。我们系统地测试了我们的常规荧光显微镜的各种组件,以优化其用于长期生物发光成像。我们的显微镜设置获得了来自活神经元的高分辨率生物发光图像,该图像可以连续拍摄数小时,没有光毒性的迹象。通过常规的发光成像仪,还可以无创地穿过完整的颅骨对小鼠大脑的生物发光进行成像。这些方法展示了如何以经济有效的方式,使用常见的试剂和设备,从活细胞和动物中常规地检测和测量生物发光。