Abstract by Jacob Vance
Chemistry and Biochemistry
Obtaining multiplexed measurements in Trypanosoma brucei with the use of fluorescent staining
Trypanosomes are a group of single-celled parasites, some of which are a major cause of neglected tropical diseases (NTDs) in humans. With the objective of identifying potential targets for drug action, our group has used Trypanosoma brucei as a model of parasite metabolism. We have adapted genetically encoded fluorescence resonance energy transfer (FRET) sensor proteins to monitor levels of analytes that are relevant to glycolysis in T. brucei. We have been successful in monitoring levels of a single analyte in response to changing conditions in the cell’s environment. However, glycolysis in the parasite is a complex pathway. By staining sensor-expressing cells with multiple fluorescent dyes, we have been able to obtain multiplexed measurements of multiple analytes in a single experiment. We demonstrate that this method can be used to obtain data for four different analytes in the same population.