The potential of autofluorescence for the detection of single living cells for label-free cell sorting in microfluidic systems.
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.
Emmelkamp
Jurjen
J
Mesa+ Institute, Lab-on-a-Chip group, University of Twente, Enschede, The Netherlands. j.emmelkamp@utwente.nl
Wolbers
Floor
F
Andersson
Helene
H
Dacosta
Ralph S
RS
Wilson
Brian C
BC
Vermes
Istvan
I
van den Berg
Albert
A
D004867:Equipment Design; D004912:Erythrocytes / Q000166:cytology; D005453:Fluorescence; D006098:Granulocytes / Q000166:cytology; D006801:Humans; D047569:Laser Scanning Cytometry / Q000295:instrumentation / Q000379:methods*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008856:Microscopy, Fluorescence