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Measurement of an Analyte Concentration in Test Solution by Using Helmholtz Resonator for Biosensor Applications.

ABSTRACT: In this paper, an indirect method of measuring an analyte concentration in a test solution using the resonant frequency change of a Helmholtz resonator is proposed, using a novel architecture of Helmholtz resonator filled with two kinds of fluids (fixed fluid and test solution). Since the analyte concentration yields changes of density and sound speed of the test solution, the resonant frequency of the proposed Helmholtz resonator is affected by the analyte concentration of the test solution. From this effect, the analyte concentration of the test solution can be measured by the spectrum of acoustic resonance of the Helmholtz resonator. The experiment was done using a 3D-printed Helmholtz resonator system with an acoustic power source and detectors, which is consistent with analytical results and showed that the analyte concentration can be measured with higher sensitivity compared to conventional cantilever-type sensors. As an example application, the possibility of measuring glucose concentration of human blood was demonstrated, showing higher sensitivity and relatively low frequency range compared to previous resonance based methods.

SUBMITTER: Chen Y

PROVIDER: S-EPMC6427302 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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Measurement of an Analyte Concentration in Test Solution by Using Helmholtz Resonator for Biosensor Applications.

Chen Yugang Y Park Yong-Hwa YH

Sensors (Basel, Switzerland) 20190305 5

In this paper, an indirect method of measuring an analyte concentration in a test solution using the resonant frequency change of a Helmholtz resonator is proposed, using a novel architecture of Helmholtz resonator filled with two kinds of fluids (fixed fluid and test solution). Since the analyte concentration yields changes of density and sound speed of the test solution, the resonant frequency of the proposed Helmholtz resonator is affected by the analyte concentration of the test solution. Fr ...[more]

PMID: 30841646

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Project description:A novel solvent-exposed analyte channel, generated by F165G substitution, on the surface of green fluorescent protein (designated His(6)GFPuv/F165G) was successfully discovered by the aid of molecular modeling software (PyMOL) in conjunction with site-directed mutagenesis. Regarding the high predictive performance of PyMOL, two pore-containing mutants namely His(6)GFPuv/H148G and His(6)GFPuv/H148G/F165G were also revealed. The pore sizes of F165G, H148G, and the double mutant H148G/F165G were in the order of 4, 4.5 and 5.5 A, respectively. These mutants were subjected to further investigation on the effect of small analytes (e.g. metal ions and hydrogen peroxide) as elucidated by fluorescence quenching experiments. Results revealed that the F165G mutant exhibited the highest metal sensitivity at physiological pH. Meanwhile, the other 2 mutants lacking histidine at position 148 had lower sensitivity against Zn(2+) and Cu(2+) than those of the template protein (His(6)GFPuv). Hence, a significant role of this histidine residue in mediating metal transfer toward the GFP chromophore was proposed and evidently demonstrated by testing in acidic condition. Results revealed that at pH 6.5 the order of metal sensitivity was found to be inverted whereby the H148G/F165G became the most sensitive mutant. The dissociation constants (K(d)) to metal ions were in the order of 4.88 x 10(-6) M, 16.67 x 10(-6) M, 25 x 10(-6) M, and 33.33 x 10(-6) M for His(6)GFPuv/F165G, His(6)GFPuv, His(6)GFPuv/H148G/F165G and His(6)GFPuv/H148G, respectively. Sensitivity against hydrogen peroxide was in the order of H148G/F165G > H148G > F165G indicating the crucial role of pore diameters. However, it should be mentioned that H148G substitution caused a markedly decrease in pH- and thermo-stability. Taken together, our findings rendered the novel pore of GFP as formed by F165G substitution to be a high impact channel without adversely affecting the intrinsic fluorescent properties. This opens up a great potential of using F165G mutant in enhancing the sensitivity of GFP in future development of biosensors.

Project description:Infection diagnosis and antibiotic susceptibility testing (AST) are time-consuming and often laborious clinical practices. This paper presents a microwave-microfluidic biosensor for rapid, contactless and non-invasive device for testing the concentration and growth of Escherichia Coli (E. Coli) in medium solutions of different pH to increase the efficacy of clinical microbiology practices. The thin layer interface between the microfluidic channel and the microwave resonator significantly enhanced the detection sensitivity. The microfluidic chip, fabricated using standard soft lithography, was injected with bacterial samples and incorporated with a microwave microstrip ring resonator sensor with an operation frequency of 2.5?GHz and initial quality factor of 83 for detecting the concentration and growth of bacteria. The resonator had a coupling gap area on of 1.5?×?1.5?mm2 as of its sensitive region. The presence of different concentrations of bacteria in different pH solutions were detected via screening the changes in resonant amplitude and frequency responses of the microwave system. The sensor device demonstrated near immediate response to changes in the concentration of bacteria and maximum sensitivity of 3.4?MHz compared to a logarithm value of bacteria concentration. The minimum prepared optical transparency of bacteria was tested at an OD600 value of 0.003. The sensor's resonant frequency and amplitude parameters were utilized to monitor bacteria growth during a 500-minute time frame, which demonstrated a stable response with respect to detecting the bacterial proliferation. A highly linear response was demonstrated for detecting bacteria concentration at various pH values. The growth of bacteria analyzed over the resonator showed an exponential growth curve with respect to time and concurred with the lag-log-stationary-death model of cell growth. This biosensor is one step forward to automate the complex AST workflow of clinical microbiology laboratories for rapid and automated detection of bacteria as well as screening the bacteria proliferation in response to antibiotics.

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Measurement of an Analyte Concentration in Test Solution by Using Helmholtz Resonator for Biosensor Applications.

Publications

Measurement of an Analyte Concentration in Test Solution by Using Helmholtz Resonator for Biosensor Applications.

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