A fluorimetric nitrite biosensor with polythienothiophene-fullerene thin film detectors for on-site water monitoring.

A novel fluorimetric sensor for highly sensitive nitrite detection on the site is presented in this study. The proposed on-chip approach comprises the use of integrated polymer photodetectors to detect light from fluorescence reactions with a diaminofluorescein probe. The detectors were prepared with a heterostructured nanofilm of polythieno[3,4-b]thiophene/benzodithiophene and (6,6)-phenyl-C71-butyric-acid methyl-ester as a photoactive layer. Prior to fluorimetric detection, the quality of the spin-coated photoactive layer was characterized via nano-morphology and current-density measurements. Nitrite assays were conducted on a poly(methyl methacrylate) microchannel chip, to which polythienothiophene-C71 based detectors were aligned. Results of signal-to-noise ratio determination have indicated a detection limit below 0.55 µM, lower than the 0.1 mg L-1 maximum limit of operation in recirculating aquaculture systems for farming Atlantic salmon Salmo salar. An increase of the nitrite concentration to toxic levels may therefore be possible to detect. The fluorimetric sensor exhibited good linearity over three orders of magnitude and acceptable detection reproducibility, which confirmed its analytical value. Further tests revealed great promise of the integrated biosensor device for detecting nitrite in aquaculture-relevant samples with high precision. The approach reported hereby may provide impetus to in situ analytical tools for monitoring water quality at aquaculture facilities, the food industries or water monitoring stations.

Recent developments in optical detection technologies in lab-on-a-chip devices for biosensing applications.

The field of microfluidics has yet to develop practical devices that provide real clinical value. One of the main reasons for this is the difficulty in realizing low-cost, sensitive, reproducible, and portable analyte detection microfluidic systems. Previous research has addressed two main approaches for the detection technologies in lab-on-a-chip devices: (a) study of the compatibility of conventional instrumentation with microfluidic structures, and (b) integration of innovative sensors contained within the microfluidic system. Despite the recent advances in electrochemical and mechanical based sensors, their drawbacks pose important challenges to their application in disposable microfluidic devices. Instead, optical detection remains an attractive solution for lab-on-a-chip devices, because of the ubiquity of the optical methods in the laboratory. Besides, robust and cost-effective devices for use in the field can be realized by integrating proper optical detection technologies on chips. This review examines the recent developments in detection technologies applied to microfluidic biosensors, especially addressing several optical methods, including fluorescence, chemiluminescence, absorbance and surface plasmon resonance.

A cascade-like silicon filter for improved recovery of oocysts from environmental waters.

Standard filtration methods have been characterized by poor recoveries when processing large-volume samples of environmental water. A method to pre-remove particulates present in turbid waters would be necessary to enhance recovery of protozoan oocysts. Particulate separation can be achieved by the proposed multiplex particle refining (MPR) system. This system employs multiple counter-flow microfiltration units that are arranged into a cascade-like structure. By use of this design, the target oocysts are pre-concentrated from environmental waters. The performance of the MPR system was investigated using 10-L deionized water and surface water spiked with 100 Cryptosporidium parvum oocysts. A recovery rate of around 85% was obtained for spiked river water. The water samples were processed using high flow rate and a simple filtration protocol. Further experiments were conducted using the MPR as a pre-filter for five commercially available filters. The recovery rates were two- to threefold higher employing the pre-filter than using the filters alone. The merit of the refining system to use different numbers of counter-flow units led to superior oocyst recovery rate for the Filta-Max and Envirochek HV filters, which are approved by the US Environmental Protection Agency. This work demonstrates a feasible tool for improved filtration performance in environmental waters.

Recovery of Cryptosporidium and Giardia organisms from surface water by counter-flow refining microfiltration.

As waterborne parasitic cryptosporidiosis and giardiasis outbreaks continue globally, monitoring of Cryptosporidium parvum and Giardia lamblia in surface water continues to be challenging. Lack of non-clogging and high-efficiency methods for recovery of C. parvum oocysts and G. lamblia cysts in environmental water strongly limits the sensitivity of detection methods for these protozoan organisms. In this work, the Counter-Flow Micro-Refinery (CFMR) system was developed by employing the novel counter-flow microfiltration principle to enrich (oo)cysts for subsequent analytical purposes. The CFMR system was constructed with multiple counter-flow concentration units that were arranged into two refining levels. By use of different numbers of units, the CFMR offered an adjustable concentration ratio allowing the concentration of 10 L and 100 L to hundreds of mL with no recirculation processing. With spiked samples, recovery of 81.3% oocysts and 86.2% cysts at a variance of < 7% was achieved for concentrations as low as 0.5-100 organisms L(-1). The recovery efficiency showed consistent for a wide range of water turbidities as well as different sample volumes. No significant clogging has been observed in the experiments. Moreover, the refining filter was able to enrich and separate oocysts and cysts in water, simultaneously. This work verifies a feasible solution for recovering C. parvum oocysts and G. lamblia cysts in large-volume surface waters. The refining system has potential to be a high-efficiency monitoring tool when combined with proper analytical detection methods.

Immunodetection of salivary biomarkers by an optical microfluidic biosensor with polyethylenimine-modified polythiophene-C70 organic photodetectors.

This work reports a novel optical microfluidic biosensor with highly sensitive organic photodetectors (OPDs) for absorbance-based detection of salivary protein biomarkers at the point of care. The compact and miniaturized biosensor has comprised OPDs made of polythiophene-C70 bulk heterojunction for the photoactive layer; whilst a calcium-free cathode interfacial layer, made of linear polyethylenimine, was incorporated to the photodetectors to enhance the low cost. The OPDs realized onto a glass chip were aligned to antibody-functionalized chambers of a poly(methyl methacrylate) microfluidic chip, in where immunogold-silver assays were conducted. The biosensor has detected IL-8, IL-1ß and MMP-8 protein in spiked saliva with high detection specificity and short analysis time exhibiting detection limits between 80pgmL-1 and 120pgmL-1. The result for IL-8 was below the clinical established cut-off of 600pgmL-1, which revealed the potential of the biosensor to early detection of oral cancer. The detection limit was also comparable to other previously reported immunosensors performed with bulky instrumentation or using inorganic photodetectors. The optical detection sensitivity of the polythiophene-C70 OPD was enhanced by optimizing the thickness of the photoactive layer and anode interfacial layer prior to the saliva immunoassays. Further, the biosensor was tested with unspiked human saliva samples, and the results of measuring IL-8 and IL-1ß were in statistical agreement with those provided by two commercial assays of ELISA. The optical microfluidic biosensor reported hereby offers an attractive and cost-effective tool to diagnostics or screening purposes at the point of care.

Ultrasensitive opto-microfluidic immunosensor integrating gold nanoparticle-enhanced chemiluminescence and highly stable organic photodetector.

The expensive fabrication of current opto-microfluidic sensors is a barrier to the successful adoption of these devices in point-of-care testing. This work reports a simple inexpensive opto-microfluidic device incorporating a poly(dimethylsiloxane)-glass hybrid microfluidic chip modified with gold nanoparticles and a high-detectivity, high-stability organic photodetector. The enhancing effect of the gold nanoparticles on horseradish peroxidase-luminol-H2O2 chemiluminescence was exploited in rapid single-analyte immunoassays. The limit of detection for 17-ß estradiol was 2.5 pg/ml, which is ∼200 times more sensitive than previously reported chemiluminescent immunosensors employing other organic photodetectors. Detection was also demonstrated in complex media, including natural water and blood serum.

Integrated optical microfluidic biosensor using a polycarbazole photodetector for point-of-care detection of hormonal compounds.

A picogram-sensitive optical microfluidic biosensor using an integrated polycarbazole photodiode is developed. The photodetector is mainly composed of the blend heterojunction of poly [N-9'-heptadecanyl-2,7-carbazole-alt-5,5-(4',7'-di-2-thienyl-2',1',3'-benzothiadiazole)] (PCDTBT) and [6,6]-phenyl C71-butyric acid methyl ester (PC70BM) and the poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) as the hole transport layer. Analyte detection is accomplished via a chemiluminescent immunoassay performed in a poly(dimethylsiloxane)-gold-glass hybrid microchip, on which antibodies were immobilized and chemiluminescent horseradish peroxidase-luminol-peroxide reactions were generated. Enhanced sensor response to the chemiluminescent light is achieved by optimizing the thickness of PCDTBT: PC70BM and PEDOT:PSS. Using the optimized polycarbazole photodiode for detecting the human thyroid-stimulating hormone as the model target, the integrated biosensor demonstrates an excellent linearity in the range of 0.03 to 10 ng/ml with an analytical sensitivity of 68 pg/ml. The sensor response shows high specificity and reproducibility. Hormone detection in clinical samples is further demonstrated and compared with a commercial enzyme-linked immunosorbent assay. The integrated device reported here has potential to detect other hormonal compounds or protein targets. Moreover, the presented concept enables the development of miniaturized, low-cost but highly sensitive optical microfluidic biosensors based on integrated polymer photodetectors with high potential for point-of-care diagnostics.

WITHDRAWN: Multiplex particle refining system to enhance Cryptosporidium recovery for surface water filtration methods.

The Publisher regrets that this article is an accidental duplication of an article that has already been published, http://dx.org/doi:10.1016/j.mimet.2012.12.009. The duplicate article has therefore been withdrawn.