Automated Sample Preparation in a Microfluidic Culture Device for Cellular Metabolomics
| Autor: | Laura A. Filla, Robert T. Filla, James L. Edwards, Katherine L. Sanders |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Lysis
Materials science Microfluidics Nanotechnology 02 engineering and technology 01 natural sciences Biochemistry Article Analytical Chemistry Specimen Handling Metabolomics Fabrication methods Lab-On-A-Chip Devices Electrochemistry Environmental Chemistry Animals Sample preparation Spectroscopy Cells Cultured Electroporation 010401 analytical chemistry Microfluidic Analytical Techniques 021001 nanoscience & nanotechnology 0104 chemical sciences Microelectrode Cattle 0210 nano-technology Microelectrodes |
| Popis: | Sample pretreatment in conventional cellular metabolomics entails rigorous lysis and extraction steps which increase the duration as well as limit the consistency of these experiments. We report a biomimetic cell culture microfluidic device (MFD) which is coupled with an automated system for rapid, reproducible cell lysis using a combination of electrical and chemical mechanisms. In-channel microelectrodes were created using facile fabrication methods, enabling the application of electric fields up to 1000 V cm(-1). Using this platform, average lysing times were 7.12 s and 3.03 s for chips with no electric fields and electric fields above 200 V cm(-1), respectively. Overall, the electroporation MFDs yielded a ∼10-fold improvement in lysing time over standard chemical approaches. Detection of multiple intracellular nucleotides and energy metabolites in MFD lysates was demonstrated using two different MS platforms. This work will allow for the integrated culture, automated lysis, and metabolic analysis of cells in an MFD which doubles as a biomimetic model of the vasculature. |
| Databáze: | OpenAIRE |
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