"Getting the best sensitivity from on-capillary fluorescence detection in capillary electrophoresis" - A tutorial

Victor A Galievsky; Alexander S Stasheuski; Sergey N Krylov

doi:10.1016/j.aca.2016.06.015

"Getting the best sensitivity from on-capillary fluorescence detection in capillary electrophoresis" - A tutorial

Anal Chim Acta. 2016 Sep 7:935:58-81. doi: 10.1016/j.aca.2016.06.015. Epub 2016 Jun 20.

Authors

Victor A Galievsky¹, Alexander S Stasheuski¹, Sergey N Krylov²

Affiliations

¹ Department of Chemistry and Centre for Research on Biomolecular Interactions, York University, Toronto, Ontario M3J 1P3, Canada.
² Department of Chemistry and Centre for Research on Biomolecular Interactions, York University, Toronto, Ontario M3J 1P3, Canada. Electronic address: skrylov@yorku.ca.

PMID: 27543015
DOI: 10.1016/j.aca.2016.06.015

Abstract

Capillary electrophoresis with Laser-Induced Fluorescence (CE-LIF) detection is being applied to new analytical problems which challenge both the power of CE separation and the sensitivity of LIF detection. On-capillary LIF detection is much more practical than post-capillary detection in a sheath-flow cell. Therefore, commercial CE instruments utilize solely on-capillary CE-LIF detection with a Limit of Detection (LOD) in the nM range, while there are multiple applications of CE-LIF that require pM or lower LODs. This tutorial analyzes all aspects of on-capillary LIF detection in CE in an attempt to identify means for improving LOD of CE-LIF with on-capillary detection. We consider principles of signal enhancement and noise reduction, as well as relevant areas of fluorophore photochemistry and fluorescent microscopy.

Keywords: Capillary electrophoresis; Highly-sensitive detection; Laser-induced fluorescence; Limit of detection; Optical filtration; Signal-to-noise ratio.

Publication types

Review