Analysis of subcellular sized particles. Capillary electrophoresis with post-column laser-induced fluorescence detection versus flow cytometry

Flow cytometry (FCM) and more recently capillary electrophoresis with post-column laser-induced fluorescence detection (CE-LIF) have both been used for subcellular particle analysis but their analytical performance has not been compared. In this work, we compare a commercial FCM with an in-house bui...

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Bibliographic Details
Published in:Journal of chromatography 2006-12, Vol.1137 (2), p.249-255
Main Authors: POE, Bobby G, NAVRATIL, Marian, ARRIAGA, Edgar A
Format: Article
Language:English
Subjects:
Aminoacridines
Analytical chemistry
Cell Fractionation - methods
Cell Line, Tumor
Chemistry
Chromatographic methods and physical methods associated with chromatography
Electrophoresis, Capillary - methods
Exact sciences and technology
Flow Cytometry - methods
Fluorescence
Humans
Lasers
Microspheres
Mitochondria - ultrastructure
Other chromatographic methods
Photobleaching
Sensitivity and Specificity
Subcellular Fractions - chemistry
Online Access:Get full text
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Summary:Flow cytometry (FCM) and more recently capillary electrophoresis with post-column laser-induced fluorescence detection (CE-LIF) have both been used for subcellular particle analysis but their analytical performance has not been compared. In this work, we compare a commercial FCM with an in-house built CE-LIF instrument using fluorescently labeled microspheres and isolated mitochondria. As evidenced by the relative standard deviation (RSD) of the individual fluorescence intensities, FCM is two-fold better than CE-LIF for microspheres with > or =1.5 x 10(6) molecules of equivalent soluble fluorescein (MESF). However, FCM has a comparatively low signal-to-noise ratio (S/N) and high RSD for microspheres with 25 times higher than FCM for all the microspheres tested and a lower RSD for microspheres with
ISSN:0021-9673
DOI:10.1016/j.chroma.2006.10.011