Cytometric analysis of protein expression and apoptosis in human primary cells with a novel microfluidic chip‐based system

Abstract

Background:Work with primary cells is inherently limited by source availability and life span in culture. Flow cytometry offers extensive analytical opportunities but generally requires high cell numbers for an experiment. Methods: We have developed assays on a microfluidic system, which allow flow cytometric analysis of apoptosis and protein expression with a minimum number of fluorescently stained primary cells. In this setup, the cells are moved by pressure-driven flow inside a network of microfluidic channels and are analyzed individually by twochannel fluorescence detection. For some assays the staining reactions can be performed on-chip and the analysis is done without further washing steps. Results:We have successfully applied the assays to evaluate (a) activation of E-selectin (CD62E) expression by

Smart Microfluidics: The Role of Stimuli- Responsive Polymers in Microfluidic Devices

“…Lee et al, 2005;Kraly et al, 2009;Abdelgawad et al, 2009) and pre-concentration,(S. Song & Singh, 2006;Anson et al, 2006) filtration and separation, (Broyles et al, 2003;Freira & Wheeler, 2006) as well as fluorescence staining,(S.D.H. Chan et al, 2003;Buhlmann et al, 2003) chemical processing and analysis. (Sims & Allbritton, 2007) A great variety of microchip fabrication techniques and materials are available for producing highly sophisticated two-and three-dimensional microstructures with integrated modules.…”

Microfluidics: Emerging prospects for anti-cancer drug screening

“…A number of emerging, microfluidic technologies for cell-based assays have recently been reported [13,14] . For instance, microfluidics offers an exceptional evolutionary route for flow cytometry, a technique known as microflow cytometry (µFCM) [27][28][29][30][31][32] . Micro fluorescently activated cell sorting (µFACS) and in-flow magnetically activated cell sorting (µMACS) are other rapidly up-andcoming examples of high-throughput on-chip cytometric technologies with substantial potential in anti-cancer drug discovery and personalized diagnostics [32][33][34][35][36] .…”

“…Micro fluorescently activated cell sorting (µFACS) and in-flow magnetically activated cell sorting (µMACS) are other rapidly up-andcoming examples of high-throughput on-chip cytometric technologies with substantial potential in anti-cancer drug discovery and personalized diagnostics [32][33][34][35][36] . Microfluidic flow cytometers and cell sorters require a greatly reduced number of cells per sample when compared with conventional FACS [27][28][29][30][31][32][33][34][35][36] . Spectral impendence using the Coulter principle has also been adapted for on chip devices to study the function of cell size, cytoplasmic resistance and membrane capacitance [37][38][39] .…”

Microfluidic devices for the analysis of apoptosis

“…Calipter technologies/Agilent has developed an annexin-V assay on microfluidic system, which allows flow cytometric analysis of apoptosis with a minimum number of cells [24,25]. In this setup, the cells are moved by pressure-driven flow inside a network of microfluidic channels and are analyzed individually by two channel fluores- …”