Biotechnological applications of bioluminescence and chemiluminescence (original) (raw)
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Bio- and chemiluminescence in bioanalysis
Fresenius' Journal of Analytical Chemistry, 2000
Analytical chemiluminescence and bioluminescence represent a versatile, ultrasensitive tool with a wide range of applications in diverse fields such as biotechnology, pharmacology, molecular biology, clinical and environmental chemistry. Enzyme activities and enzyme substrates and inhibitors can be efficiently determined when directly involved in luminescent reactions, and also when they take part in a reaction suitable for coupling to a final light-emitting reaction. Chemiluminescence detection has been exploited in the fields of flow-injection analysis and column-liquid chromatographic and capillary-electrophoretic separative systems, due to its high sensitivity when compared with colorimetric detection. It has widely been used as an indicator of reactive oxygen species formation in cells and whole organs, thus allowing the study of a number of pathophysiological conditions related to oxidative stress. Chemiluminescence represents a sensitive and rapid alternative to radioactivity as a detection principle in immunoassays for the determination of a wide range of molecules (hormones, food additives, environmental pollutants) and in filter membrane biospecific reactions (Southern, Northern, Western, dot blot) for the determination of nucleic acids and proteins. Chemiluminescence has also been used for the sensitive and specific localization and quantitation of target analytes in tissue sections and single cells by immunohistochemistry and in situ hybridization techniques. A relatively recent application regards the use of luminescent reporter genes for the development of bioassays based on genetically engineered microorganisms or mammalian cells able to emit visible light in response to specific inorganic and organic compounds. Finally, the high detectability and rapidity of bio-and chemiluminescent detection make it suitable for the development of microarraybased high throughput screening assays, in which simultaneous, multianalyte detection is performed on multiple samples.
Luminescence, 2020
The recent advancements in synthetic biology, organic chemistry and computational models, allowed the application of bioluminescence in several fields, ranging from well-established methods for detecting microbial contamination to in vivo imaging to track cancer and stem cells, from cell-based assays to optogenetics. Moreover, thanks to recent technological progresses in miniaturized and sensitive light detectors, such as photodiodes and imaging sensors, it is possible to implement laboratory-based assays, such as cell-based and enzymatic assays, into portable analytical devices for point-of-care and on-site applications. This review highlights some recent advances in the development of whole-cell and cell-free bioluminescent biosensors with a glance on current challenges and different strategies that were used to turn bioassays into biosensors with the required analytical performance. Critical issues and unsolved technical problems are also highlighted, to give the reader a taste of this fascinating and challenging field.
Journal of Histochemistry & Cytochemistry, 1993
The breakthrough of chemiluminescence in the field of solution immunoassays and transfer membranes prompted us to explore whether a light-based detection system could provide a gain in sensitivity over chromogenic and FITC markers for nucleic acid and protein detection on histological preparations. A Hamamatsu device and an enhanced chemiluminescence (ECL) luminol substrate of the peroxidase were used to detect epithelial and endothelial components by immunohistochemistry (IHC) and for in situ hybridization (ISH) of papilloma virus DNA. The accuracy of the signal was compared to that obtained with DAB-peroxidase, silver-enhanced DAB-peroxidase, NBT-BCIP-alkaline phosphatase, and FITC. Our results demonstrated the feasibility and high sensitivity of luminescence detection for histological preparations. In part due to the ultrasensitive videocamera and photon-counting imaging, interpretable and reproducible results were obtained within counting times shorter than 5 min, and with dilut...
Progress in chemical luminescence-based biosensors: A critical review
Biosensors and Bioelectronics, 2016
Biosensors are a very active research field. They have the potential to lead to low-cost, rapid, sensitive, reproducible, and miniaturized bioanalytical devices, which exploit the high binding avidity and selectivity of biospecific binding molecules together with highly sensitive detection principles. Of the optical biosensors, those based on chemical luminescence detection (including chemiluminescence, bioluminescence, electrogenerated chemiluminescence, and thermochemiluminescence) are particularly attractive, due to their high-to-signal ratio and the simplicity of the required measurement equipment. Several biosensors based on chemical luminescence have been described for quantitative, and in some cases multiplex, analysis of organic molecules (such as hormones, drugs, pollutants), proteins, and nucleic acids. These exploit a variety of miniaturized analytical formats, such as microfluidics, microarrays, paper-based analytical devices, and whole-cell biosensors. Nevertheless, despite the high analytical performances described in the literature, the field of chemical luminescence biosensors has yet to demonstrate commercial success. This review presents the main recent advances in the field and discusses the approaches, challenges, and open issues, with the aim of stimulating a broader interest in developing chemical luminescence biosensors and improving their commercial exploitation.
Method for Implementing Bioluminescence-Based Analytical Assays in Nanoliter Volumes
Bioluminescence and Chemiluminescence - Progress and Perspectives - Proceedings of the 13th International Symposium, 2005
Bioluminescence-based analytical assays were used to measure various analytes in nanoliter sample volumes. Nanoliter volumes of multiple bioluminescent analytical assays were deposited in an array format and lyophilized. ATP-firefly luciferase (FFL) and NADH-bacterial luciferase (BL) platform reactions were compared. We achieved parallel sample delivery via sample-hydrated membranes. A CCD camera measured the luminescent kinetics for each assay. These miniaturized assays and instruments can be prepared as micro-analytical systems to operate in point-of-care (POC) diagnostic devices.
Towards portable, real-time, integrated fluorescence microarray diagnostics tools
IRBM, 2007
We present recent results on biochips slides with high fluorescence efficiency and their associated readout systems. To obtain the full performance made possible by such slides, the chemical surface functionalization has to be improved, as formerly unobservable defects of the functionalization are now made observable through the improved efficiency. The systems are based on the integration of the hybridization and readout functions into a single machine instead the usual two separate systems, quite cumbersome and expensive. Ultimate performance is reached with systems using standard imaging circuits, CCD or CMOS, as the biochip substrate. In this case, one can obtain a remarkable miniaturization of the full optical system and the integrated hybridization chamber/readout head can be reduced to the size of a webcam.
Bio- and chemiluminescence imaging in analytical chemistry
Analytica Chimica Acta, 2005
Bio-and chemiluminescence imaging techniques combine the high sensitivity of bio-and chemiluminescence detection with the ability of current light imaging devices to localize and quantify light emission down to the single-photon level. These techniques have been successfully exploited for the development of sensitive analytical methods relying on the evaluation of the spatial distribution of the light emitted from a target sample. In this paper, we report on recent applications of bio-and chemiluminescence imaging for in vitro and in vivo assays, including: quantitative assays performed in various analytical formats, such as microtiter plates, microarrays and miniaturized analytical devices, used in the pharmaceutical, clinical, diagnostic and environmental fields; luminescence imaging microscopy based on enzymatic, immunohistochemical and in situ hybridization reactions for the localization of metabolites, enzymes, antigens and gene sequences in cells and tissues; whole-body luminescence imaging in live animals for evaluating biological and pathological processes and for pharmacological studies.
Sensors, 2009
Whole-cell, genetically modified bioreporters are designed to emit detectable signals in response to a target analyte or related group of analytes. When integrated with a transducer capable of measuring those signals, a biosensor results that acts as a self-contained analytical system useful in basic and applied environmental, medical, pharmacological, and agricultural sciences. Historically, these devices have focused on signaling proteins such as green fluorescent protein, aequorin, firefly luciferase, and/or bacterial luciferase. The biochemistry and genetic development of these sensor systems as well as the advantages, challenges, and common applications of each one will be discussed.
The publication is divided into two volumes. Volume I (Springer Vol. 1571) focuses on optical-based detectors, while Vol. II (Springer Vol. 1572) focuses on electrochemical,bioelectronic, piezoelectric, cellular, and molecular biosensors. Volume I (Springer Vol. 1571): Optical-based detection, encompasses a broad array of technologies including direct and indirect methods as discussed above. Part I of Vol. I describes various optical-based direct detectors. Three types of direct optical detection biosensors are described: evanescent wave (SPR and resonant waveguide grating), interferometers, and Raman spectroscopy sensors. Part II focuses on indirect optical detection. Part 2 of Vol. I describes various indirect optical detectors as discussed above. Indirect directors require a labeled molecule to be bound to the signal-generating target. For optical sensors such molecules emit or modify light signals. Most indirect optical detectors are designed to measure fluorescence; however, such detectors can also measure densitometric and colorimetric changes as well as chemiluminescence, and detection depends on the type of label used. Such optical signals can be measured in various ways as described in Part II. These include various CCD-based detectors which are very versatile, inexpensive, and relatively simple to construct and use. Other optical detectors discussed in Part II are photodiode-based detection systems and mobile phone detectors. Lateral flow systems that rely on visual detection are included in this section. Although lateral flow devices are not “classical” biosensors with ligands and transducers, they are included in this book because of their importance for biosensing. Lateral flow assays use simple immunodetection (or DNA hybridization) devices, such as competitive or sandwich assays, and are used mainly for medical diagnostics such as laboratory and home testing or any other point of care (POC) detection. A common format is a “dipstick” in which the test sample flows on an absorbant matrix via capillary action; detection is accomplished by mixing a colorimetric reagent with the sample and binding to a secondary antibody to produce lines or zones at specific locations on the absorbing matrix. Volume II (Springer Vol. 1572): Volume II describes various electrochemical, bioelectronic, piezoelectric, and cellular- and molecular-based biosensors. In Part I of Vol. II, we describe several types of electrochemical and bioelectronic detectors. Electrochemical biosensors were the first biosensors developed and are the most commonly used biosensors in clinical settings (e.g., glucose monitoring). Also included are several electronic/semiconductor sensors based on the field effect. Unlike electrochemical sensors, which are indirect detectors and require labeling, electronic/semiconductor biosensors are label-free. In Part II we describe “mechanical detectors” which modify their mechanical properties as a result of biological interactions. Such mechanical direct biosensors include piezoelectric biosensors which change their acoustical resonance and cantilevers which modify their movement. Part III describes a variety of biological sensors including aptamer-based sensors and cellular and phage display technologies. Part IV describes several microfluidics technologies for cell isolation. In addition, a number of related technologies including Raman spectroscopy and high-resolution microultrasound are described. The two volumes provide comprehensive and detailed technical protocols on current biosensor and biodetection technologies and examples of their applications and capabilities.