Modeling and measurement of a whole-cell bioluminescent biosensor based on a single photon avalanche diode

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• 作者：Ramiz Daniel ; Ronen Almog ; Amit Ron ; Shimshon Belkin ; Yosi Shacahm Diam
• 关键词：Whole-cell biosensor ; Bioluminescence ; SOS response ; Single photon avalanche photodiode
• 刊名：Biosensors and Bioelectronics
• 出版年：2008
• 出版时间：1 December 2008
• 年：2008
• 卷：24
• 期：4
• 页码：888-893
• 全文大小：493 K

文摘

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 20 ppm).