An optical relay approach to very low cost hybrid polymer-complementary metal-oxide semiconductor electrophoresis instrumentation

Hall, Gordon H.; Sloan, David; Ma, Tingxuan; Couse, Madeline; Martel, Stéphane; Elliott, D.G.; Glerum, D. Moira; Backhouse, C.

doi:10.1016/j.chroma.2014.05.005

Cited by 7 publications

(13 citation statements)

References 16 publications

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“…The distance from the intersection to the edge of S was 2 mm and the distance from the intersection to the detection point was 17 mm. Further detail can be found in the supporting information and has been previously reported .…”

Section: Methodsmentioning

confidence: 98%

“…Separations were performed in 2.5% agarose in a 0.5 x running buffer. Further detail can be found in the supporting information and in past work .…”

Section: Methodsmentioning

confidence: 99%

“…The resulting electropherograms were processed with a standard data processing algorithm . The algorithm first discounts all preseparation signal.…”

Section: Methodsmentioning

confidence: 99%

“…The emission pathway was based upon an optical relay arrangement in which two GRIN lenses (GRIN2306A, Thorlabs) image the photodiode active area (SFH2701, OSRAM Semiconductor) onto the bottom of the chip. The GRIN lens arrangement provides a high light collection efficiency due to their high numerical aperture and proximity to the channel . In contrast to most other systems, the optically probed volume was limited horizontally by a 200 μm pinhole in the emission pathway rather than placing an aperture on the excitation path.…”

Section: Methodsmentioning

confidence: 99%

“…Since low power LEDs were used, photobleaching of the sample was not apparent, unlike the situation commonly found with higher power LED or laser systems. In the vertical direction, the light collection efficiency drops substantially for distances greater than 100 μm away from the chip bottom . The emission filter was a 3 mm thick piece of amber plastic (Acrylite 2C04, Evonik).…”

Section: Methodsmentioning

confidence: 99%

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Light emitting diode, photodiode‐based fluorescence detection system for DNA analysis with microchip electrophoresis

2015

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Electrophoretic separation of fluorescently end-labeled DNA after a PCR serves as a gold standard in genetic diagnostics. Because of their size and cost, instruments for this type of analysis have had limited market uptake, particularly for point-of-care applications. This might be changed through a higher level of system integration and lower instrument costs that can be realized through the use of LEDs for excitation and photodiodes for detection--if they provide sufficient sensitivity. Here, we demonstrate an optimized microchip electrophoresis instrument using polymeric fluidic chips with fluorescence detection of end-labeled DNA with a LOD of 0.15 nM of Alexa Fluor 532. This represents orders of magnitude improvement over previously reported instruments of this type. We demonstrate the system with an electrophoretic separation of two PCR products and their respective primers. We believe that this is the first LED-induced fluorescence microchip electrophoresis system with photodiode-based detection that could be used for standard applications of PCR and electrophoresis.

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Section: Methodsmentioning

confidence: 98%

“…Separations were performed in 2.5% agarose in a 0.5 x running buffer. Further detail can be found in the supporting information and in past work .…”

Section: Methodsmentioning

confidence: 99%

“…The resulting electropherograms were processed with a standard data processing algorithm . The algorithm first discounts all preseparation signal.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Light emitting diode, photodiode‐based fluorescence detection system for DNA analysis with microchip electrophoresis

2015

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Effective length calibration method for processing the fluorescence signal detected by charge‐coupled device in capillary electrophoresis

Liu

Jin

et al. 2017

J of Separation Science

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A novel method named effective length calibration method has been developed to process the fluorescence signal detected by charge-coupled device during capillary electrophoresis. The new method treated each pixel as an individual point detector, and effectively binned a large number of pixels into a final electropherogram without losing the narrow detection window defined by a single pixel. Capillary electrophoresis separations of DNA were carried out and detected by charge-coupled device and conventional detector (photomultiplier tube). Detection properties including signal-to-noise ratio, peak width, detection frequency, and tilt of detector were investigated. It was found that the new method achieved much higher signal-to-noise ratio and smaller peak width than the conventional detector did. A Detection width of 0.5 μm was easily achieved.

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Study of the peak broadening due to detection in the electrophoretic separation of DNA by CE and microchip CE and the application of image sensor for ultra‐small detection cell length

Zhao

Chen

et al. 2019

J of Separation Science

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Narrow peaks are important to high-resolution and high-speed separation of DNA fragments by capillary electrophoresis and microchip capillary electrophoresis.Detection cell length is one of the broadening factors, which is often ignored in experiments. However, is it always safe to neglect detection cell length under any condition? To answer this question, we investigated the influence of detection cell length by simulation and experiments. A parameter named as detection cell length ratio was proposed to directly compare the detection cell length and the spatial length of sample band. Electrophoretic peaks generated by various detection cell length ratios were analyzed. A simple rule to evaluate the peak broadening due to detection cell length was obtained. The current states of the detection cell length of detection system and their reliabilities in capillary electrophoresis and microchip capillary electrophoresis were analyzed. Microchip capillary electrophoresis detection with an ultra-small detection cell length of 0.36 μm was easily achieved by using an image sensor. K E Y W O R D Scharge-coupled-device, detection cell length, microchip capillary electrophoresis, peak broadening 2280 Study of the peak broadening due to detection in the electrophoretic separation of DNA by CE and microchip CE and the application of image sensor for ultra-small detection cell length.

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An optical relay approach to very low cost hybrid polymer-complementary metal-oxide semiconductor electrophoresis instrumentation

Cited by 7 publications

References 16 publications

Light emitting diode, photodiode‐based fluorescence detection system for DNA analysis with microchip electrophoresis

Light emitting diode, photodiode‐based fluorescence detection system for DNA analysis with microchip electrophoresis

Effective length calibration method for processing the fluorescence signal detected by charge‐coupled device in capillary electrophoresis

Study of the peak broadening due to detection in the electrophoretic separation of DNA by CE and microchip CE and the application of image sensor for ultra‐small detection cell length

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