Light-Based Detection Methods for Capillary Electrophoresis

Lapainis, Theodore; Sweedler, Jonathan V.; Scanlan, Cory

doi:10.1201/9781420004953.ch9

Cited by 2 publications

(2 citation statements)

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“…Coupled to CE, fluorescence detection provides extremely sensitive limits of detection, particularly when detecting brightly fluorescent molecules (5) . LIF requires either on-column or post-column excitation with a laser of appropriate wavelength, and efficient collection and measurement of fluorescence emission.…”

Section: Fluorescence Detectionmentioning

confidence: 99%

“…The gravity-driven sheath flow matches or slightly exceeds the flow in the capillary, minimising turbulence and focussing the analyte stream, reducing background from the cuvette walls (5,25) . A solid-state laser, matched to the optimal excitation of the derivatized analytes, is fibre-coupled to a 10Â objective and focussed just below the capillary output.…”

Section: Coupling Ce-lif To a Microbeammentioning

confidence: 99%

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Microbeam-coupled capillary electrophoresis

Garty

Ehsan

Yang

et al. 2015

Radiat Prot Dosimetry

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Section: Fluorescence Detectionmentioning

confidence: 99%

Section: Coupling Ce-lif To a Microbeammentioning

confidence: 99%

Microbeam-coupled capillary electrophoresis

Garty

Ehsan

Yang

et al. 2015

Radiat Prot Dosimetry

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Improvement of LOD in Fluorescence Detection with Spectrally Nonuniform Background by Optimization of Emission Filtering

2017

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The limit-of-detection (LOD) in analytical instruments with fluorescence detection can be improved by reducing noise of optical background. Efficiently reducing optical background noise in systems with spectrally nonuniform background requires complex optimization of an emission filter-the main element of spectral filtration. Here, we introduce a filter-optimization method, which utilizes an expression for the signal-to-noise ratio (SNR) as a function of (i) all noise components (dark, shot, and flicker), (ii) emission spectrum of the analyte, (iii) emission spectrum of the optical background, and (iv) transmittance spectrum of the emission filter. In essence, the noise components and the emission spectra are determined experimentally and substituted into the expression. This leaves a single variable-the transmittance spectrum of the filter-which is optimized numerically by maximizing SNR. Maximizing SNR provides an accurate way of filter optimization, while a previously used approach based on maximizing a signal-to-background ratio (SBR) is the approximation that can lead to much poorer LOD specifically in detection of fluorescently labeled biomolecules. The proposed filter-optimization method will be an indispensable tool for developing new and improving existing fluorescence-detection systems aiming at ultimately low LOD.

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Light-Based Detection Methods for Capillary Electrophoresis

Cited by 2 publications

References 0 publications

Microbeam-coupled capillary electrophoresis

Microbeam-coupled capillary electrophoresis

Improvement of LOD in Fluorescence Detection with Spectrally Nonuniform Background by Optimization of Emission Filtering

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