Quasi-Confocal, Multichannel Parallel Scan Hyperspectral Fluorescence Imaging Method Optimized for Analysis of Multicolor Microarrays

Shi

et al. 2011

Sensors

Self Cite

Conventional fluorescence scanners utilize multiple filters to distinguish different fluorescent labels, and problems arise because of this filter-based mechanism. In this work we propose a line-monitoring, hyperspectral fluorescence technique which is designed and optimized for applications in multi-channel microfluidic systems. In contrast to the filter-based mechanism, which only records fluorescent intensities, the hyperspectral technique records the full spectrum for every point on the sample plane. Multivariate data exploitation is then applied to spectra analysis to determine ratios of different fluorescent labels and eliminate unwanted artifacts. This sensor is designed to monitor multiple fluidic channels simultaneously, providing the potential for multi-analyte biosensing. The detection sensitivity is approximately 0.81 fluors/μm2, and this sensor is proved to act with a good homogeneity. Finally, a model experiment of detecting short oligonucleotides has demonstrated the biomedical application of this hyperspectral fluorescence biosensor.

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Line-Monitoring, Hyperspectral Fluorescence Setup for Simultaneous Multi-Analyte Biosensing

Shi

et al. 2011

Sensors

Self Cite

“…To detect multiple mutations, melting curves of multiple samples are analyzed by using similar analyzers to those used for qPCR. 6,7 As for nucleic-acid-hybridization-based analysis on a microarray plate 8,9 or a microtiter plate, 10 multi-ple target DNA or RNA fragments are immobilized and independently detected. As for an antigen-antibody-reaction-based immunoassay on a microarray plate 11,12 or a microtiter plate, 13 multiple target proteins are immobilized and independently detected.…”

Section: Introductionmentioning

confidence: 99%

An ultra-small, multi-point, and multi-color photo-detection system with high sensitivity and high dynamic range

Anazawa

Yamazaki

2017

Lab Chip

Although multi-point, multi-color fluorescence-detection systems are widely used in various sciences, they would find wider applications if they are miniaturized. Accordingly, an ultra-small, four-emission-point and four-color fluorescence-detection system was developed. Its size (space between emission points and a detection plane) is 15 × 10 × 12 mm, which is three-orders-of-magnitude smaller than that of a conventional system. Fluorescence from four emission points with an interval of 1 mm on the same plane was respectively collimated by four lenses and split into four color fluxes by four dichroic mirrors. Then, a total of sixteen parallel color fluxes were directly input into an image sensor and simultaneously detected. The emission-point plane and the detection plane (the image-sensor surface) were parallel and separated by a distance of only 12 mm. The developed system was applied to four-capillary array electrophoresis and successfully achieved Sanger DNA sequencing. Moreover, compared with a conventional system, the developed system had equivalent high fluorescence-detection sensitivity (lower detection limit of 17 pM dROX) and 1.6-orders-of-magnitude higher dynamic range (4.3 orders of magnitude).

“…Our group has developed this technology by using the method of quasi-confocal and multichannel parallel scanning to analyse multicolor microarrays. The advanced method has exhibited great potential for applications in microarrays [22]. In our previous work [23], we proposed SPR-supporting sensing structures based on symmetrical CPWR and the experimental results showed that the sensing characteristics of this symmetrical CPWR based sensor are greatly improved over conventional SPR sensors (a refractive index resolution of 8.1 × 10 −8 RIU and 3.5 × 10 −7 RIU in water and line on the sensor films to cover all the channels of the microfluidic system for fluorescence excitation, and thus multiple channels can be monitored simultaneously at a dark field.…”

Section: Introductionmentioning

confidence: 99%

Multi-Channel Hyperspectral Fluorescence Detection Excited by Coupled Plasmon-Waveguide Resonance

Zhang

et al. 2013

Sensors

Self Cite

We propose in this paper a biosensor scheme based on coupled plasmon-waveguide resonance (CPWR) excited fluorescence spectroscopy. A symmetrical structure that offers higher surface electric field strengths, longer surface propagation lengths and depths is developed to support guided waveguide modes for the efficient excitation of fluorescence. The optimal parameters for the sensor films are theoretically and experimentally investigated, leading to a detection limit of 0.1 nM (for a Cy5 solution). Multiplex analysis possible with the fluorescence detection is further advanced by employing the hyperspectral fluorescence technique to record the full spectra for every pixel on the sample plane. We demonstrate experimentally that highly overlapping fluorescence (Cy5 and Dylight680) can be distinguished and ratios of different emission sources can be determined accurately. This biosensor shows great potential for multiplex detections of fluorescence analytes.