Measuring Dynamic Glycosomal pH Changes in Living <i>Trypanosoma brucei</i>

Daniel Call; Sabrina S. Pizarro; Erica Tovey; Emily Knight; Carrie Baumgardner; Kenneth A. Christensen; James C. Morris

doi:10.3791/66279

Journal

Immunology and Infection

Development of pHluorin2-Based Sensor for Glycosomal pH in Trypanosoma brucei

Development of pHluorin2-Based Sensor for Glycosomal pH in Trypanosoma brucei

January 19, 2024

Author Spotlight: Advancements in Glycosomal pH Monitoring in Trypanosoma brucei Using pHluorin2 Biosensor

01:46 min

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272 Views

Sample Preparation for Flow Cytometry to Measure pHluorin2 Fluorescence in Living Trypanosoma brucei

00:54 min

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34 Views

Flow Cytometry Procedure to Measure Relative pH of the Glycosome in Living Trypanosoma brucei

02:15 min

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20 Views

Analysis of Flow Cytometry Data to Measure the Relative pH of the Glycosome in Living Trypanosoma brucei

04:00 min

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22 Views

Daniel Call*¹, Sabrina S. Pizarro*^2,3, Erica Tovey¹, Emily Knight^2,3, Carrie Baumgardner^2,4, Kenneth A. Christensen¹, James C. Morris²

¹Department Chemistry and Biochemistry,Brigham Young University, ²Eukaryotic Pathogens Innovation Center,Clemson University, ³Department of Genetics and Biochemistry,Clemson University, ⁴Department of Physics and Astronomy,Clemson University

Summary

We describe a method to study how pH responds to environmental cues in the glycosomes of the bloodstream form of African trypanosomes. This approach involves a pH-sensitive heritable protein sensor in combination with flow cytometry to measure pH dynamics, both as a time-course assay and in a high-throughput screen format.