title: A new tool for routine testing of cellular protein expression: integration of cell staining and analysis of protein expression on a microfluidic chip-based system.
abstract: The key benefits of Lab-on-a-Chip technology are substantial time savings via an automation of lab processes, and a reduction in sample and reagent volumes required to perform analysis. In this article we present a new implementation of cell assays on disposable microfluidic chips. The applications are based on the controlled movement of cells by pressure-driven flow in microfluidic channels and two-color fluorescence detection of single cells. This new technology allows for simple flow cytometric studies of cells in a microfluidic chip-based system. In addition, we developed staining procedures that work "on-chip," thus eliminating time-consuming washing steps. Cells and staining-reagents are loaded directly onto the microfluidic chip and analysis can start after a short incubation time. These procedures require only a fraction of the staining reagents generally needed for flow cytometry and only 30,000 cells per sample, demonstrating the advantages of microfluidic technology. The specific advantage of an on-chip staining reaction is the amount of time, cells, and reagents saved, which is of great importance when working with limited numbers of cells, e.g., primary cells or when needing to perform routine tests of cell cultures as a quality control step. Applications of this technology are antibody staining of proteins and determination of cell transfection efficiency by GFP expression. Results obtained with microfluidic chips, using standard cell lines and primary cells, show good correlation with data obtained using a conventional flow cytometer.
authors.affiliation: Agilent Technologies Deutschland GmbH, Hewlett-Packard Strasse 8, D76337Waldbronn, Germany. Carsten_Buhlmann@Agilent.com
authors.forename: Carsten
authors.initials: C
authors.lastname: Buhlmann
authors.forename: Tobias
authors.initials: T
authors.lastname: Preckel
authors.forename: Samuel
authors.initials: S
authors.lastname: Chan
authors.forename: Gerd
authors.initials: G
authors.lastname: Luedke
authors.forename: Marc
authors.initials: M
authors.lastname: Valer
mesh_terms: D000906:Antibodies; D015703:Antigens, CD / Q000276:immunology; D001331:Automation; D051940:B7-2 Antigen; D005434:Flow Cytometry; D020869:Gene Expression Profiling / Q000379:methods*; D008562:Membrane Glycoproteins / Q000276:immunology; D044085:Microfluidics; D040901:Proteomics / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000906:Antibodies; D015703:Antigens, CD; D051940:B7-2 Antigen; D008562:Membrane Glycoproteins