Kostov Yordan, Sergeev Nikolay, Wilson Sean, Herold Keith E, Rasooly Avraham
Center for Advanced Sensor Technology, University of Maryland Baltimore County, Baltimore, MD, USA.
Methods Mol Biol. 2009;503:259-72. doi: 10.1007/978-1-60327-567-5_14.
In this chapter we describe a simple and relatively inexpensive Electroluminescence (EL) illumination and charged-coupled device (CCD) camera (EL-CCD) based detector for monitoring fluorescence and colorimetric assays. The portable battery-operated fluorescence detector includes an EL panel for fluoro-genic excitation at 490 nm, a cooled CCD digital camera to monitor emission at 523 nm, filters and a close up lens. The detector system is controlled by a laptop computer for camera operation, image acquisition and analysis. The system was tested using a fluorogenic peptide substrate (SNAP-25) for botulinum neurotoxin serotype A (BoNT-A) labeled with FITC. The level of detection of the system was found to be 1.25 nM of the peptide, similar to the detection level of a commercial photomultipler-based plate fluorometer. The multichannel EL-CCD was used with an assay plate capable of testing nine samples simultaneously in 1 min at this detection level. The portable system is small and is operated by a 12 V source. The modular detector was designed with easily interchangeable ELs, filters and lenses and can be used and adapted for a wide variety of fluorescence and colorimetric assays, fluorescence labels and assay formats.
在本章中,我们描述了一种基于电致发光(EL)照明和电荷耦合器件(CCD)相机(EL-CCD)的简单且相对廉价的检测器,用于监测荧光和比色测定。这种便携式电池供电的荧光检测器包括一个用于在490nm进行荧光激发的EL面板、一个用于监测523nm发射的冷却CCD数码相机、滤光片和一个近摄镜头。该检测系统由一台笔记本电脑控制,用于相机操作、图像采集和分析。使用一种用异硫氰酸荧光素(FITC)标记的肉毒杆菌神经毒素A(BoNT-A)的荧光肽底物(SNAP-25)对该系统进行了测试。发现该系统的检测水平为该肽的1.25nM,与基于商业光电倍增管的平板荧光计的检测水平相似。在这个检测水平下,多通道EL-CCD与一个能够在1分钟内同时测试9个样品的检测板一起使用。该便携式系统体积小,由12V电源供电。该模块化检测器的设计使得EL、滤光片和镜头易于互换,可用于并适用于各种荧光和比色测定、荧光标记和检测形式。