title: MicroPrep: chip-based dielectrophoretic purification of mitochondria.
abstract: We have developed a microfluidic system--microPrep--for subcellular fractionation of cell homogenates based on dielectrophoretic sorting. Separation of mitochondria isolated from a human lymphoblastoid cell line was monitored by fluorescence microscopy and further characterized by western blot analysis. Robust high throughput and continuous long-term operation for up to 60 h of the microPrep chip system with complex biological samples became feasible as a result of a comprehensive set of technical measures: (i) coating of the inner surfaces of the chip with BSA, (ii) application of mechanical actuators to induce periodic flow patterns, (iii) efficient cooling of the device to ensure integrity of organelle, (iv) a wide channel to provide for high fluidic throughput, and (v) integration of a serial arrangement of 10 dielectrophoretic deflector units to enable separation of samples with a high particle load without clogging. Hence, microPrep yields tens of micrograms of enriched and purified mitochondria within hours. Western blots of mitochondria fractions showed that contaminating endoplasmatic reticulum was reduced by a factor 6 when compared with samples prepared by state of the art centrifugation.
authors.affiliation: Natural and Medical Sciences Institute at the University of Tübingen, Reutlingen, Germany.
authors.forename: Meike
authors.initials: M
authors.lastname: Moschallski
authors.forename: Monika
authors.initials: M
authors.lastname: Hausmann
authors.forename: Anton
authors.initials: A
authors.lastname: Posch
authors.forename: Aran
authors.initials: A
authors.lastname: Paulus
authors.forename: Nancy
authors.initials: N
authors.lastname: Kunz
authors.forename: Thanh Tu
authors.initials: TT
authors.lastname: Duong
authors.forename: Brigitte
authors.initials: B
authors.lastname: Angres
authors.forename: Kai
authors.initials: K
authors.lastname: Fuchsberger
authors.forename: Heiko
authors.initials: H
authors.lastname: Steuer
authors.forename: Dieter
authors.initials: D
authors.lastname: Stoll
authors.forename: Simon
authors.initials: S
authors.lastname: Werner
authors.forename: Britta
authors.initials: B
authors.lastname: Hagmeyer
authors.forename: Martin
authors.initials: M
authors.lastname: Stelzle
mesh_terms: D002458:Cell Fractionation / Q000295:instrumentation / Q000379:methods*; D045744:Cell Line, Tumor; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D006801:Humans; D008928:Mitochondria / Q000737:chemistry*; D024101:Mitochondrial Proteins / Q000032:analysis*
chemical_list: D024101:Mitochondrial Proteins