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The potential of autofluorescence for the detection of single living cells for label‐free cell sorting in microfluidic systems
Author(s) -
Emmelkamp Jurjen,
Wolbers Floor,
Andersson Helene,
DaCosta Ralph S.,
Wilson Brian C.,
Vermes Istvan,
van den Berg Albert
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200406070
Subject(s) - microfluidics , autofluorescence , cell sorting , single cell analysis , fluorescence , fluorescence microscope , materials science , flow cytometry , confocal , nanotechnology , microscope , confocal microscopy , cell , microscopy , biomedical engineering , chemistry , microbiology and biotechnology , biology , optics , medicine , biochemistry , physics , genetics
A novel method for studying unlabeled living mammalian cells based on their autofluorescence (AF) signal in a prototype microfluidic device is presented. When combined, cellular AF detection and microfluidic devices have the potential to facilitate high‐throughput analysis of different cell populations. To demonstrate this, unlabeled cultured cells in microfluidic devices were excited with a 488 nm excitation light and the AF emission (> 505 nm) was detected using a confocal fluorescence microscope (CFM). For example, a simple microfluidic three‐port glass microstructure was used together with conventional electroosmotic flow (EOF) to switch the direction of the fluid flow. As a means to test the potential of AF‐based cell sorting in this microfluidic device, granulocytes were successfully differentiated from human red blood cells (RBCs) based on differences in AF. This study demonstrated the use of a simple microfabricated device to perform high‐throughput live cell detection and differentiation without the need for cell‐specific fluorescent labeling dyes and thereby reducing the sample preparation time. Hence, the combined use of microfluidic devices and cell AF may have many applications in single‐cell analysis.