Journal
ELECTROPHORESIS
Volume 25, Issue 21-22, Pages 3513-3527Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/elps.200406086
Keywords
capillary electrophoresis; detection limit; laser-induced fluorescence; microchip electrophoresis; review
Funding
- NINDS NIH HHS [F33NS046828, R15NS038443] Funding Source: Medline
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Laser-induced fluorescence is an extremely sensitive method for detection in chemical separations. In addition, it is well-suited to detection in small volumes, and as such is widely used for capillary electrophoresis and microchip-based separations. This review explores the detailed instrumental conditions required for sub-zeptomole, sub-picomolar detection limits. The key to achieving the best sensitivity is to use an excitation and emission volume that is matched to the separation system and that, simultaneously, will keep scattering and luminescence background to a minimum. We discuss how this is accomplished with confocal detection, 90degrees on-capillary detection, and sheath-flow detection. It is shown that each of these methods have their advantages and disadvantages, but that all can be used to produce extremely sensitive detectors for capillary- or microchip-based separations. Analysis of these capabilities allows prediction of the optimal means of achieving ultrasensitive detection on microchips.
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