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JoVE Journal

Immunology and Infection

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Measuring the pH, Redox Chemistries, and Degradative Capacity of Macropinosomes using Dual-Fluorophore Ratiometric Microscopy

2.3K Views

07:31 min

August 19th, 2021

August 19th, 2021

2,283 Views

0:05

Introduction

0:28

Measuring Macropinosome pH

2:06

In Situ Calibration of Macropinosome pH

3:04

Measuring Oxidative Events within Macropinosomes

4:22

Measuring Protein Digestion within Macropinosomes

5:30

Results: Measuring the pH, Oxidative Events, and Protein Degradation in Macropinosomes using Dual-Fluorophore Ratiometric Microscopy

6:59

Conclusion

Transcript

This protocol allows for the real-time assessment of various processes and biochemical parameters within the lumen of individual macropinosomes. These methods can be employed to drug discovery in the cell biology of macropinocytosis. This protocol

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We describe protocols for measuring pH, oxidative events, and protein digestion in individual macropinosomes in live cells. An emphasis is placed on dual-fluorophore ratiometric microscopy and the advantages it offers over population-based techniques.

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