Denis R. Santos

A point-of-use microfluidic device with integrated photodetector array for immunoassay multiplexing: Detection of a panel of mycotoxins in multiple samples.

For a point-of-use analytical device to be successful in real-world applications, it needs to be rapid, simple to operate and, ideally, able to multiplex the detection of several analytes and samples. Mycotoxin detection in food and feedstock in particular has become increasingly relevant as these toxins, such as ochratoxin A (OTA), aflatoxin B1 (AFB1) and deoxynivalenol (DON), are subject to strict regulations and recommendations in the European Union. A novel, simple, negative pressure-driven device with manually operated magnetic valves was developed and the simultaneous immunodetection of these three mycotoxins was demonstrated via the laminar flow patterning of probes in an area of ≈0.12mm2 and subsequent chemiluminescence generation via HRP-labeled antibodies. The three mycotoxins were detected in less than 20min at concentrations of 100ng/mL for OTA and DON and 3ng/mL for AFB1, spiked in a sample under analysis and simultaneously compared to a toxin-free reference and a standard contaminated with critical target concentrations. The on-chip optical detection was performed in a single acquisition step by integrating a microfabricated array of 25×25µm2 hydrogenated amorphous silicon (a-Si:H) photosensors below the microfluidic chip. The device presented in this work is simple and effective for point-of-use multiplexing of immunoassays and was applied in this work to the screening of mycotoxins.

Performance of Hydrogenated Amorphous Silicon Thin Film Photosensors at Ultra-Low Light Levels: Towards Attomole Sensitivities in Lab-on-Chip Biosensing Applications

Thin-film optical transducers integrated in lab-on-a-chip (LoC) devices have the potential to facilitate miniaturization, multiplexing capabilities, portability, and sensitivity. However, there are few systematic studies that provide detailed characterization of these photosensors to particular miniaturized bioassays at reduced light intensities. A more detailed representation of the photosensors performance at low light levels is thus needed in order to improve the engineering of integrated detectors for the next generation of portable biosensors. Here, an assessment of the performance of hydrogenated amorphous silicon (a-Si:H) photosensors, at ultra-low light intensity, based on two different device configurations for integration in a microfluidic platform for biomolecular detection is presented. Both p-i-n photodiodes and parallel contact photoconductors show low dark current density (~10−10 A.cm−2) and photosensitivity comparable with high performing crystalline silicon photosensors. The a-Si:H photosensors were integrated in microfluidic devices for the detection of antibody-horseradish peroxidase conjugates (Ab-HRP) in a chemiluminescence-based assay. Surface concentrations of Ab-HRP as low as ~26 amol.cm−2 and ~9 fmol.cm−2 (corresponding to ~103 and ~106 HRP molecules) were detected with detectors in photodiode and photoconductor configurations, respectively, with the sensitivity of the former being comparable with that of bulky and ultra-sensitive photomultiplier tubes. The excellent sensitivity and dynamic range obtained, together with the miniaturization and arraying potential, highlight the potential of a-Si:H photosensors as an effective means of integrating the multiplexed acquisition of optical data in real time in point-of-need LoC systems.