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 florescence and colorimetric assays. The portable battery-operated florescence 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 florescence and colorimetric assays, florescence labels and assay formats.
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Acknowledgments
We thank Mr. Steven Sun and Mr. Jesse Frances for their technical assistance. This work was supported in part by the Office of Public Health emergency Preparedness (OPHEP) IAG 224-05-655 (to A. Rasooly and by FDA contract HHSF223200610765P (to Dr. Yordan Kostov)
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Kostov, Y., Sergeev, N., Wilson, S., Herold, K.E., Rasooly, A. (2009). A Simple Portable Electroluminescence Illumination-Based CCD Detector. In: Rasooly, A., Herold, K.E. (eds) Biosensors and Biodetection. Methods in Molecular Biology™, vol 503. Humana Press. https://doi.org/10.1007/978-1-60327-567-5_14
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