TY - JOUR
T1 - A cell array biosensor for environmental toxicity analysis
AU - Jin, Hyung Lee
AU - Mitchell, Robert J.
AU - Byoung, Chan Kim
AU - Cullen, David C.
AU - Man, Bock Gu
N1 - Funding Information:
The authors would like to thank Dr. He Sun Chung and Mr. Yoon-Shik Nam at KIST for kindly allowing using CCD camera. This research was supported by the National Research Laboratory (NRL) Program of Korea Institute of Science and Technology Evaluation and Planning (Project No. M10318000094-04J0000-02710). We are grateful for the support.
Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2005/9/15
Y1 - 2005/9/15
N2 - In this study, a cell-based array technology that uses recombinant bioluminescent bacteria to detect and classify environmental toxicity has been implemented to develop two biosensor arrays, i.e., a chip and a plate array. Twenty recombinant bioluminescent bacteria, having different promoters fused with the bacterial lux genes, were immobilized within LB-agar. About 2 μl of the cell-agar mixture was deposited into the wells of either a cell chip or a 384-well plate. The bioluminescence (BL) from the cell arrays was measured with the use of highly sensitive cooled CCD camera that measured the bioluminescent signal from the immobilized cells and then quantified the pixel density using image analysis software. The responses from the cell arrays were characterized using three chemicals that cause either superoxide damage (paraquat), DNA damage (mitomycin C) or protein/membrane damage (salicylic acid). The responses were found to be dependent upon the promoter fused upstream of the lux operon within each strain. Therefore, a sample's toxicity can be analyzed and classified through the changes in the BL expression from each well. Moreover, a time of only 2 h was needed for analysis, making either of these arrays a fast, portable and economical high-throughput biosensor system for detecting environmental toxicities.
AB - In this study, a cell-based array technology that uses recombinant bioluminescent bacteria to detect and classify environmental toxicity has been implemented to develop two biosensor arrays, i.e., a chip and a plate array. Twenty recombinant bioluminescent bacteria, having different promoters fused with the bacterial lux genes, were immobilized within LB-agar. About 2 μl of the cell-agar mixture was deposited into the wells of either a cell chip or a 384-well plate. The bioluminescence (BL) from the cell arrays was measured with the use of highly sensitive cooled CCD camera that measured the bioluminescent signal from the immobilized cells and then quantified the pixel density using image analysis software. The responses from the cell arrays were characterized using three chemicals that cause either superoxide damage (paraquat), DNA damage (mitomycin C) or protein/membrane damage (salicylic acid). The responses were found to be dependent upon the promoter fused upstream of the lux operon within each strain. Therefore, a sample's toxicity can be analyzed and classified through the changes in the BL expression from each well. Moreover, a time of only 2 h was needed for analysis, making either of these arrays a fast, portable and economical high-throughput biosensor system for detecting environmental toxicities.
KW - Bioluminescent bacteria
KW - Cell array chip
KW - Environmental biosensor
KW - Toxicity analysis
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U2 - 10.1016/j.bios.2004.12.015
DO - 10.1016/j.bios.2004.12.015
M3 - Article
C2 - 16076440
AN - SCOPUS:23044515022
SN - 0956-5663
VL - 21
SP - 500
EP - 507
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
IS - 3
ER -