Fluorescence imaging of the activity of glucose oxidase using a hydrogen-peroxide-sensitive europium probe.
A method for optical imaging of the activity of glucose oxidase (GOx) using a fluorescent europium(III) tetracycline probe for hydrogen peroxide is presented. A decay time in the microsecond range and the large Stokes shift of 210 nm of the probe facilitate intensity-based, time-resolved, and decay-time-based imaging of glucose oxidase. Four methods for imaging the activity of GOx were compared, and rapid lifetime determination imaging was found to be the best in giving a linear range from 0.32 to 2.7 m Unit/mL. The detection limit is 0.32 m Unit/mL (1.7 ng mL(-1)) which is similar to that of the time-resolved (gated) imaging using a microtiterplate reader. Fluorescent imaging of the activity of GOx is considered to be a useful tool for GOx-based immunoassays with potential for high-throughput screening, immobilization studies, and biosensor array technologies.
Wu
Meng
M
Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, D-93040 Regensburg, Germany.
Lin
Zhihong
Z
Schäferling
Michael
M
Dürkop
Axel
A
Wolfbeis
Otto S
OS
D015374:Biosensing Techniques; D005063:Europium / Q000737:chemistry*; D005949:Glucose Oxidase / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000032:analysis* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D007700:Kinetics; D015335:Molecular Probes / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013752:Tetracycline / Q000737:chemistry; D013997:Time Factors
D015335:Molecular Probes; D005063:Europium; D006861:Hydrogen Peroxide; D005949:Glucose Oxidase; D013752:Tetracycline