Lab-on-a-chip technologies for proteomic analysis from isolated cells

Abstract

Lab-on-a-chip systems offer a versatile environment in which low numbers of cells and molecules can be manipulated, captured, detected and analysed. We describe here a microfluidic device that allows the isolation, electroporation and lysis of single cells. A431 human epithelial carcinoma cells, expressing a green fluorescent protein-labelled actin, were trapped by dielectrophoresis within an integrated lab-on-a-chip device containing saw-tooth microelectrodes. Using these same trapping electrodes, on-chip electroporation was performed, resulting in cell lysis. Protein release was monitored by confocal fluorescence microscopy.

Microfluidic electroporation for cellular analysis and delivery

“…A common set of electrode pattern can be implemented to initially trap and concentrate target cells in regions of interest by dielectrophoretic force and then perform subsequent electroporation while cells are exposed to electric pulsing protocols. 60, 67–69, 72–76, 78–80, 82, 83, 89 Therefore, this technique, adopted in either static or flow-through manner, is particularly suitable for applications combining electrical cell lysis with downstream intracellular content analysis 72–76, 79, 80, 82 .…”

“…Techniques other than electrophoresis were also used for analysis of intracellular molecules released by electroporation. 58, 67–70, 79, 80, 104, 123, 124 An integrated device was reported on F 1 -ATPase rotational assay at the single molecule level. 80 In the work, genetically engineered E. coli expressing 6× His-tag fusion F 1 -ATPase was continuously loaded into the chip and initially lysed by interdigitated sawtooth planar microelectrodes in a chamber.…”

Microscale electroporation: challenges and perspectives for clinical applications

“…Also, the overall process of protein release can be monitored by confocal fluorescence microscopy. 61 To use relatively low voltages in single-cell electroporation, electrophoresis can be used to enhance delivery of compounds, followed by electroporation. For example, this can be used to assist electroporative uptake of impermanent molecules such as Calcein (MW: 622 Da) and Oregon Green Dextran (MW: 70 000 Da) in microchannels.…”

Microtechnology meets systems biology: The small molecules of metabolome as next big targets

“…Single cell and single molecular analytic methods may bring about breakthrough in metabolic research in the future (Amantonico et al, 2009;Krylov et al, 2000;Andersson and van den Berg, 2004;Sedgwick et al, 2008). Many more burdens have to be overcome before they can be really applied to metabolic analysis of real samples from physiologically defined cultures, especially from processes of biotechnology.…”

“…Up to now, the application of microfluidic systems in biotechnology rather focus on the fields of genomic or proteomic research, on-chip cell culturing and the integration of micro analytical techniques (Shendure and Ji, 2008;Andersson and van den Berg, 2003;Sedgwick et al, 2008;Weibel et al, 2007). Usually, the systems are optimized towards minimum sample volume and are designed to meet the requirements for serial production and disposability.…”