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Immunology and Infection

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

Article DOI: 10.3791/62733-v • 07:31 min • August 19th, 2021
August 19th, 2021 •
1,2, 2,3
1Department of Biochemistry & Microbiology, Faculty of Science, University of Victoria, 2Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Science, University of Calgary, 3Calvin, Joan and Phoebe Snyder Institute for Chronic Diseases, University of Calgary

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résumé
Automatic Translation

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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PH Measurement Redox Chemistries Degradative Capacity Macropinosomes Dual-fluorophore Ratiometric Microscopy Real-time Assessment Biochemical Parameters Drug Discovery Cell Biology Heterogeneity Raw264.7 Cells Glass Bottom 96-well Plate HBSS TMR Labeled Dextran Fluorescein Labeled Dextran Incubation Wash Steps Microscope Imaging
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