Encapsulation, Manipulation, and Analysis of Single Cells in Individual Compartments: A Functional FACS Device in a Microfluidic Chip
Zusammenfassung der Projektergebnisse
A modular microfluidic chip system was developed which enabled us to encapsulate cells individually in picoliter-sized aqueous drops in an inert oil phase, incubate, and manipulate the cells while they are still in the drops, and retrieve them from the drops alive. By choosing the right microchannel geometry, flow parameters, oil phase, and surfactants we ensured that the cells could be efficiently encapsulated and kept alive in the drops for sufficient time spans. These techniques are valuable whenever cells are to be studied on a single cell level yet in large numbers to obtain relevant statistical information. High-throughput screening assays such as screening of cells for specific properties (e.g. production of a specific antibody or secretion of certain proteins) can be performed in a time- and cost-efficient manner. We were able to demonstrate the utility of the modular concept by encapsulating hybridoma cells and measuring the concentration of the antibodies which they produced while incubated on chip. Furthermore, a microfluidic drop trapping device was designed to store drops which can, for example, contain individual cells. The drops can be kept in place over long periods of time and the contents of the drops can be observed by microscopic or spectroscopic methods. We were able to keep blood platelets in suspension inside confining drops over several hours and observe their highly dynamic behavior even in the non-activated state. The same device was used to co-encapsulate bacteria and bacteriophages and enabled us to observe cells lysis as a consequence of virus infection as a function of time and in dependence of relevant parameters such as virus genome length and solution salt concentration. Drop-based microfluidics is a powerful technique both from a biotechnological standpoint as well as a tool to investigate fundamental biochemical and biophysical question in cellular systems. The techniques developed and applied in this project will be applicable for many more technological and scientific questions.
Projektbezogene Publikationen (Auswahl)
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Biocompatible surfactants for water-in-fluorocarbon emulsion. Lab on a Chip
Christian Holtze, Amy C. Rowat, Jeremy J. Agresti, J. Brian Hutchison, Francesco Elio Angile, Christian H.W. Schmitz, Sarah Köster, Honey Duan, Katherine J. Humphry, Randall A. Scanga, Jeff S. Johnson, Dario Pisignano, and David A. Weitz
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Dropspots: a picoliter array in a microfluidic device. Lab on a Chip
Christian H.W. Schmitz, Amy C. Rowat, Sarah Köster, and David A. Weitz
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Controlled encapsulation of single cells into monodisperse picolitre drops. Lab on a Chip 8 (2008), 1262 – 1264
Jon F. Edd, Dino Di Carlo, Katherine J. Humphry, Sarah Köster, Daniel Irimia, David A. Weitz, and Mehmet Toner
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Drop-based microfluidic devices for encapsulation of single cells. Lab on a Chip 8 (2008), 1110 – 1115
Sarah Köster, Francesco E. Angilè, Honey Duan, Anton Wintner, Jeremy J. Agresti, Amy C. Rowat, Christoph A. Merten, Dario Pisignano, Andrew D. Griffiths, and David A. Weitz
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Droplet-Based Microfluidic Platforms for the Encapsulation and Screening of Mammalian Cells and Multicellular Organisms. Chemistry and Biology 15 (2008), 427-437
Jenifer Clausell-Tormos, Diana Lieber, Abdeslam El-Harrak, Oliver J. Miller, Jean-Christophe Baret, Joshua Blouwolff, Katie Humphrey, Sarah Köster, Honey Duan, Christian Holtze, David A. Weitz, Andrew D. Griffiths, and Christoph A. Merten
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Internal Capsid-Pressure Dependence of Virus Infection by Phage λ. Physical Review Letters
Sarah Köster, Meerim Jeembaeva, Alex Evilevitch, and David A. Weitz
