Intracellular Manipulation of Phagosomal Transport and Maturation Using Magnetic Tweezers

Shashank Shekhar, Vinod Subramaniam, Johannes S Kanger

    Research output: Chapter in Book / Report / Conference proceedingChapterAcademic

    Abstract

    Phagocytosis is an important process of the immune system by which pathogens are internalized and eliminated by phagocytic cells. Upon internalization, the phagosome matures and acidifies while being transported in a centripetal fashion. In this chapter, we describe protocols for simultaneous imaging of phagosomal acidification as well as their spatial manipulation by magnetic tweezers. First, we describe the protocols for functionalization of magnetic microbeads with pH-sensitive dyes and pH calibration of these particles. We also describe the preparation of magnetic tweezers and the calibration of forces that can be generated by these tweezers. We provide details of the design of the custom electrical and optical setup used for simultaneous imaging of phagosomal pH and phagosome's location. Finally, we provide a detailed description of the data analysis methodology.

    Original languageEnglish
    Title of host publicationPhagocytosis and Phagosomes
    Subtitle of host publicationMethods and Protocols
    EditorsRobert Botelho
    PublisherSpringer
    Pages93-112
    Number of pages20
    ISBN (Electronic)9781493965816
    ISBN (Print)9781493965793, 9781493982417
    DOIs
    Publication statusPublished - 2017

    Publication series

    NameMethods in Molecular Biology
    PublisherHumana Press
    Volume1519
    ISSN (Print)1064-3745

    Keywords

    • Journal Article

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