Chapter title |
Intracellular Manipulation of Phagosomal Transport and Maturation Using Magnetic Tweezers.
|
---|---|
Chapter number | 7 |
Book title |
Phagocytosis and Phagosomes
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6581-6_7 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6579-3, 978-1-4939-6581-6
|
Authors |
Shashank Shekhar, Vinod Subramaniam, Johannes S. Kanger |
Editors |
Roberto Botelho |
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. |
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Geographical breakdown
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Unknown | 5 | 100% |
Demographic breakdown
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Researcher | 2 | 40% |
Student > Bachelor | 1 | 20% |
Other | 1 | 20% |
Student > Postgraduate | 1 | 20% |
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Neuroscience | 1 | 20% |
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Unknown | 1 | 20% |