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Measuring in vivo protein dynamics throughout the cell cycle using microfluidics. / de Leeuw, Roy; Brazda, Peter; Moolman, M. Charl; Kerssemakers, J. W.J.; Solano Hermosilla, B.P.; Dekker, Nynke H.

The Bacterial Nucleoid : methods and Protocols. ed. / Olivier Espéli. Vol. 1624 Humana Press, 2017. p. 237-252 (Methods in Molecular Biology; Vol. 1624).

Research output: Chapter in Book/Report/Conference proceedingChapterScientific

Harvard

de Leeuw, R, Brazda, P, Moolman, MC, Kerssemakers, JWJ, Solano Hermosilla, BP & Dekker, NH 2017, Measuring in vivo protein dynamics throughout the cell cycle using microfluidics. in O Espéli (ed.), The Bacterial Nucleoid : methods and Protocols. vol. 1624, Methods in Molecular Biology, vol. 1624, Humana Press, pp. 237-252. https://doi.org/10.1007/978-1-4939-7098-8_18

APA

de Leeuw, R., Brazda, P., Moolman, M. C., Kerssemakers, J. W. J., Solano Hermosilla, B. P., & Dekker, N. H. (2017). Measuring in vivo protein dynamics throughout the cell cycle using microfluidics. In O. Espéli (Ed.), The Bacterial Nucleoid : methods and Protocols (Vol. 1624, pp. 237-252). (Methods in Molecular Biology; Vol. 1624). Humana Press. https://doi.org/10.1007/978-1-4939-7098-8_18

Vancouver

de Leeuw R, Brazda P, Moolman MC, Kerssemakers JWJ, Solano Hermosilla BP, Dekker NH. Measuring in vivo protein dynamics throughout the cell cycle using microfluidics. In Espéli O, editor, The Bacterial Nucleoid : methods and Protocols. Vol. 1624. Humana Press. 2017. p. 237-252. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-7098-8_18

Author

de Leeuw, Roy ; Brazda, Peter ; Moolman, M. Charl ; Kerssemakers, J. W.J. ; Solano Hermosilla, B.P. ; Dekker, Nynke H. / Measuring in vivo protein dynamics throughout the cell cycle using microfluidics. The Bacterial Nucleoid : methods and Protocols. editor / Olivier Espéli. Vol. 1624 Humana Press, 2017. pp. 237-252 (Methods in Molecular Biology).

BibTeX

@inbook{17c4448ae86042569851e5c1880bf0a8,
title = "Measuring in vivo protein dynamics throughout the cell cycle using microfluidics",
abstract = "Studying the dynamics of intracellular processes and investigating the interaction of individual macromolecules in live cells is one of the main objectives of cell biology. These macromolecules move, assemble, disassemble, and reorganize themselves in distinct manners under specific physiological conditions throughout the cell cycle. Therefore, in vivo experimental methods that enable the study of individual molecules inside cells at controlled culturing conditions have proved to be powerful tools to obtain insights into the molecular roles of these macromolecules and how their individual behavior influence cell physiology. The importance of controlled experimental conditions is enhanced when the investigated phenomenon covers long time periods, or perhaps multiple cell cycles. An example is the detection and quantification of proteins during bacterial DNA replication. Wide-field microscopy combined with microfluidics is a suitable technique for this. During fluorescence experiments, microfluidics offer well-defined cellular orientation and immobilization, flow and medium interchangeability, and high-throughput long-term experimentation of cells. Here we present a protocol for the combined use of wide-field microscopy and microfluidics for the study of proteins of the Escherichia coli DNA replication process. We discuss the preparation and application of a microfluidic device, data acquisition steps, and image analysis procedures to determine the stoichiometry and dynamics of a replisome component throughout the cell cycle of live bacterial cells.",
keywords = "DNA replication, Escherichia coli, Fluorescence imaging, Microfluidics, Single-molecule techniques",
author = "{de Leeuw}, Roy and Peter Brazda and Moolman, {M. Charl} and Kerssemakers, {J. W.J.} and {Solano Hermosilla}, B.P. and Dekker, {Nynke H.}",
year = "2017",
doi = "10.1007/978-1-4939-7098-8_18",
language = "English",
isbn = "978-1493970971",
volume = "1624",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "237--252",
editor = "Olivier Esp{\'e}li",
booktitle = "The Bacterial Nucleoid",

}

RIS

TY - CHAP

T1 - Measuring in vivo protein dynamics throughout the cell cycle using microfluidics

AU - de Leeuw, Roy

AU - Brazda, Peter

AU - Moolman, M. Charl

AU - Kerssemakers, J. W.J.

AU - Solano Hermosilla, B.P.

AU - Dekker, Nynke H.

PY - 2017

Y1 - 2017

N2 - Studying the dynamics of intracellular processes and investigating the interaction of individual macromolecules in live cells is one of the main objectives of cell biology. These macromolecules move, assemble, disassemble, and reorganize themselves in distinct manners under specific physiological conditions throughout the cell cycle. Therefore, in vivo experimental methods that enable the study of individual molecules inside cells at controlled culturing conditions have proved to be powerful tools to obtain insights into the molecular roles of these macromolecules and how their individual behavior influence cell physiology. The importance of controlled experimental conditions is enhanced when the investigated phenomenon covers long time periods, or perhaps multiple cell cycles. An example is the detection and quantification of proteins during bacterial DNA replication. Wide-field microscopy combined with microfluidics is a suitable technique for this. During fluorescence experiments, microfluidics offer well-defined cellular orientation and immobilization, flow and medium interchangeability, and high-throughput long-term experimentation of cells. Here we present a protocol for the combined use of wide-field microscopy and microfluidics for the study of proteins of the Escherichia coli DNA replication process. We discuss the preparation and application of a microfluidic device, data acquisition steps, and image analysis procedures to determine the stoichiometry and dynamics of a replisome component throughout the cell cycle of live bacterial cells.

AB - Studying the dynamics of intracellular processes and investigating the interaction of individual macromolecules in live cells is one of the main objectives of cell biology. These macromolecules move, assemble, disassemble, and reorganize themselves in distinct manners under specific physiological conditions throughout the cell cycle. Therefore, in vivo experimental methods that enable the study of individual molecules inside cells at controlled culturing conditions have proved to be powerful tools to obtain insights into the molecular roles of these macromolecules and how their individual behavior influence cell physiology. The importance of controlled experimental conditions is enhanced when the investigated phenomenon covers long time periods, or perhaps multiple cell cycles. An example is the detection and quantification of proteins during bacterial DNA replication. Wide-field microscopy combined with microfluidics is a suitable technique for this. During fluorescence experiments, microfluidics offer well-defined cellular orientation and immobilization, flow and medium interchangeability, and high-throughput long-term experimentation of cells. Here we present a protocol for the combined use of wide-field microscopy and microfluidics for the study of proteins of the Escherichia coli DNA replication process. We discuss the preparation and application of a microfluidic device, data acquisition steps, and image analysis procedures to determine the stoichiometry and dynamics of a replisome component throughout the cell cycle of live bacterial cells.

KW - DNA replication

KW - Escherichia coli

KW - Fluorescence imaging

KW - Microfluidics

KW - Single-molecule techniques

UR - http://www.scopus.com/inward/record.url?scp=85028616308&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-7098-8_18

DO - 10.1007/978-1-4939-7098-8_18

M3 - Chapter

SN - 978-1493970971

VL - 1624

T3 - Methods in Molecular Biology

SP - 237

EP - 252

BT - The Bacterial Nucleoid

A2 - Espéli, Olivier

PB - Humana Press

ER -

ID: 33977738