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Optical Quantification of Intracellular pH in Drosophila melanogaster Malpighian Tubule Epithelia with a Fluorescent Genetically-encoded pH Indicator
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Biologia
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JoVE Journal Biologia
Optical Quantification of Intracellular pH in Drosophila melanogaster Malpighian Tubule Epithelia with a Fluorescent Genetically-encoded pH Indicator

Optical Quantification of Intracellular pH in Drosophila melanogaster Malpighian Tubule Epithelia with a Fluorescent Genetically-encoded pH Indicator

DOI:
10.3791/55698-v

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11:54 min

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August 11, 2017

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,
1Department of Physiology and Biomedical Engineering,Mayo Clinic College of Medicine, 2Department of Nephrology and Hypertension,Mayo Clinic College of Medicine

Capitoli

  • 00:05Titolo
  • 00:54Dissection of Adult Drosophila Anterior Malpighian Tubules
  • 04:32Validation of Imaging Protocol and Tubule Health
  • 09:47Results: Quantification of Acid Extrusion in Malpighian Tubule Epithelia
  • 10:56Conclusion

Summary

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Cellular ion transport can often be assessed by monitoring intracellular pH (pHi). Genetically Encoded pH-Indicators (GEpHIs) provide optical quantification of intracellular pH in intact cells. This protocol details the quantification of intracellular pH through cellular ex vivo live-imaging of Malpighian tubules of Drosophila melanogaster with pHerry, a pseudo-ratiometric genetically encoded pH-indicator.

Tags

Intracellular PHDrosophila MelanogasterMalpighian Tubule EpitheliaFluorescent Genetically-encoded PH IndicatorBasolateral Proton TransportEpithelial Proton MovementPH RegulationInsect EpithelialMammalian NephronEpithelial CellsIPBSSchneider’s MediumDissectionForceps

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