title: Modeling and measurement of a whole-cell bioluminescent biosensor based on a single photon avalanche diode.
abstract: Whole-cell biosensors are potential candidates for on-line and in situ environmental monitoring. In this work we present a new design of a whole-cell bioluminescence biosensor for water toxicity detection, based on genetically engineered Escherichia coli bacteria, carrying a recA::luxCDABE promoter-reporter fusion. Sensitive optical detection is achieved using a single photon avalanche photodiode (SPAD) working in the Geiger mode. The present work describes a simple mathematical model for the kinetic process of the bioluminescence based SOS toxin response of E. coli bacteria. We find that initially the bioluminescence signal depends on the time square and we show that the spectral intensity of the bioluminescence signal is inverse proportional to the frequency. We get excellent agreement between the theoretical model and the measured light signal. Furthermore, we present experimental results of the bioluminescent signal measurement using a SPAD and a photomultiplier, and demonstrate improvement of the measurement by applying a matched digital filter. Low intensity bioluminescence signals were measured after the whole-cell sensors were exposed to various toxicant concentrations (5, 15 and 20ppm).
authors.affiliation: Department of Physical Electronic, Electrical Engineering Faculty, Tel Aviv University, Ramat Aviv, 69978 Tel-Aviv, Israel.
authors.forename: Ramiz
authors.initials: R
authors.lastname: Daniel
authors.forename: Ronen
authors.initials: R
authors.lastname: Almog
authors.forename: Amit
authors.initials: A
authors.lastname: Ron
authors.forename: Shimshon
authors.initials: S
authors.lastname: Belkin
authors.forename: Yosi Shacahm
authors.initials: YS
authors.lastname: Diamand
mesh_terms: D000900:Anti-Bacterial Agents / Q000008:administration & dosage; D001681:Biological Assay / Q000295:instrumentation* / Q000379:methods; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D003198:Computer Simulation; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000187:drug effects* / Q000502:physiology*; D008163:Luminescent Measurements / Q000295:instrumentation* / Q000379:methods; D008954:Models, Biological; D009268:Nalidixic Acid / Q000008:administration & dosage*; D017785:Photons; D012666:Semiconductors
chemical_list: D000900:Anti-Bacterial Agents; D009268:Nalidixic Acid