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Measuring Phagosome pH by Ratiometric Fluorescence Microscopy
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Measuring Phagosome pH by Ratiometric Fluorescence Microscopy
DOI:

14:39 min

December 07, 2015

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Chapters

  • 00:05Title
  • 01:02Preparation of Phagocytic Targets
  • 04:18Live Video Microscopy
  • 05:47Calibration and Control Experiments
  • 06:34Analysis
  • 11:44Results: Time Courses, Average pH and Ratio Histograms of Wild-type and Hvcn1-/- Neutrophil Phagosomes
  • 13:08Conclusion

Summary

Automatic Translation

Phagosomal pH influences phagosome maturation, oxidant production, phagosomal killing as well as antigen presentation. Here we describe a ratiometric method for measuring time-course and endpoint pH changes in individual phagosomes in living phagocytes using fluorescence microscopy.

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