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Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator

Imaging the Neutrophil Phagosome and Cytoplasm Using a Ratiometric pH Indicator

DOI:
10.3791/55107-v

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12:42 min

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April 05, 2017

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, , , ,
1Centre for Molecular Medicine, Division of Medicine,University College London, 2Cell and Developmental Biology,University College London

Chapters

  • 00:05Title
  • 01:05S-1 Coupling to Heat-killed (HK) Candida
  • 03:19Isolation of Neutrophils
  • 05:20Preparation of Slides
  • 07:27Confocal Microscopy
  • 08:57Image Analysis
  • 10:06Results: Images of Neutrophils from Humans and Mice
  • 11:29Conclusion

Summary

Automatic Translation

Automatic Translation

This manuscript describes a simple method to measure the phagosomal pH and area as well as the cytoplasmic pH of human and mouse neutrophils using the ratiometric indicator seminaphthorhodafluor (SNARF)-1, or S-1. This is achieved using live-cell confocal fluorescence microscopy and image analysis.

Tags

NeutrophilPhagosomeCytoplasmPH IndicatorConfocal MicroscopyNADPH OxidasePhagocytic VacuoleIon FluxesCarboxy-S-1 Succinimidyl EsterCandidaOpsonizationHuman NeutrophilsPeripheral BloodDextran

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