A sensitive CCD image system for detection of chemiluminescent reactions

Jansen, Eugène; Buskens, C.A.F.; Berg, Rijk H. van den

doi:10.1002/bio.1170030204

Cited by 15 publications

(4 citation statements)

References 4 publications

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“…A detection limit of 0.001 amol of enzyme has been reported . Currently, the most sensitive enzymatic chemiluminescent assay is AP and the recently developed 1,2-dioxetane substrate, CSPD. − This extreme sensitivity is largely due to improved substrates and advancements in photon sensing technology such as photomultiplyer tubes (PMTs) and in charge-coupled device (CCD) cameras. − New cooled CCD cameras have very low background current, which allows the signal to be integrated over longer periods of time, thereby increasing sensitivity. Moreover, the array-based construction of CCD cameras is well suited for the simultaneous determination of multiple samples.…”

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confidence: 99%

Microformat Imaging ELISA for Pesticide Determination

et al. 1996

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A flat-well microformat competitive enzyme-linked immunosorbent chemiluminescent assay for the detection of the pesticide 2,4-dichlorophenoxyacetic acid (2,4-D) is described. Thick-film technology was used to pattern a hydrophobic layer 100 μm thick onto glass microscope slides to form an array of 2 × 2 mm(2) squares. These flat wells were able to hold 2 μL of reagents, corresponding to a height of ∼500 μm, with minimal contamination risk. The hydrophobic ink used to pattern the surfaces allowed significantly larger volumes of samples to be applied when compared with surfaces patterned with nonhydrophobic inks. This reduced evaporation effects and permitted greater pipetting accuracy, thereby improving assay reproducibility. A competitive immunoassay was developed based on the ability of free 2,4-D hapten to inhibit binding of anti-2,4-D monoclonal antibodies to 2,4-D-bovine serum albumin conjugate adsorbed onto the glass support. The support was subsequently incubated with alkaline phosphatase (AP) labeled anti-mouse IgG. The amount of AP conjugate bound was determined by quantitating the chemiluminescent emission produced from the enzymatic breakdown of CSPD substrate by AP using a cooled CCD camera. The detection limit of the single-sample microformat assay was 2.7 × 10(-)(11) M, or 6 pg of 2,4-D. The linear ranges of the single-sample and multisample assays were 4.5 × 10(-)(8)-4.5 × 10(-)(11) and 4.5 × 10(-)(7)-1.66 × 10(-)(10) M, respectively. In comparison, the detection limits of a tube-based chemiluminescent assay using standard luminometer and of a colorimetric ELISA were 45 × 10(-)(11) and 9.9 × 10(-)(8) M, respectively. The ability to scale the thick-film-based microformat assay makes it an ideal candidate for the development of affinity arrays and high-throughput assay formats. Prospects for further improvements of this imaging ELISA strategy will be discussed.

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confidence: 99%

Microformat Imaging ELISA for Pesticide Determination

et al. 1996

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“…In addition to luminol, many CL substrates, such as acridinium ester, peroxyoxalate, 1,2-dioxetane and their derivatives, have been found. With the development of CL systems and advanced analytical equipment with high resolution such as photomultiplier tube and charge-coupled device (CCD), CL imaging technology has become attractive in in vitro and in vivo applications 2, 3. To get better imaging effect, CCD-based imaging devices have become more and more popular 4.…”

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confidence: 99%

Chemiluminescence and Bioluminescence Imaging for Biosensing and Therapy: In Vitro and In Vivo Perspectives

Yan

Shi

Song³

et al. 2019

Theranostics

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Chemiluminescence (CL) and bioluminescence (BL) imaging technologies, which require no external light source so as to avoid the photobleaching, background interference and autoluminescence, have become powerful tools in biochemical analysis and biomedical science with the development of advanced imaging equipment. CL imaging technology has been widely applied to high-throughput detection of a variety of analytes because of its high sensitivity, high efficiency and high signal-to-noise ratio (SNR). Using luciferase and fluorescent proteins as reporters, various BL imaging systems have been developed innovatively for real-time monitoring of diverse molecules in vivo based on the reaction between luciferin and the substrate. Meanwhile, the kinetics of protein interactions even in deep tissues has been studied by BL imaging. In this review, we summarize in vitro and in vivo applications of CL and BL imaging for biosensing and therapy. We first focus on in vitro CL imaging from the view of improving the sensitivity. Then, in vivo CL applications in cells and tissues based on different CL systems are demonstrated. Subsequently, the recent in vitro and in vivo applications of BL imaging are summarized. Finally, we provide the insight into the development trends and future perspectives of CL and BL imaging technologies.

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“…Luminometers are central to all luminescence studies and in the past, journals have published extensive surveys of luminometers . In addition, the literature contains numerous reviews and evaluations of luminometers and descriptions of luminometers based on charge‐coupled detectors (CCD) invented in 1969 , low cost detectors (e.g., photographic film, silicon photodiodes) , single photon counting and luminometers designed for specific types of reaction vessels such as microplates or for direct injection of reagents for luminescence reactions with rapid emission kinetics .…”

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confidence: 99%