Microfluidics-based single cell analysis reveals drug-dependent motility changes in trypanosomes.

Area of Science:

• Parasitology
• Microfluidics
• Drug Discovery

Background:

• Trypanosoma brucei is a flagellated unicellular parasite causing significant human diseases.
• Assessing drug efficacy against trypanosomes requires methods that can quantify effects on motility and viability.
• Current drug screening methods may lack the precision to differentiate cytostatic from cytocidal effects in situ.

Purpose of the Study:

• To develop and validate a single-cell viability assay using microfluidics and optical micromanipulation.
• To investigate the impact of chemical agents on the motility and viability of Trypanosoma brucei in situ.
• To establish a platform for rapid, high-content drug screening against parasitic protozoa.

Main Methods:

• Integration of chemical gradient microfluidics with optical micromanipulation for real-time monitoring.
• Quantification of local cell velocity and mean squared displacement (MSD) of individual trypanosome trajectories.
• In situ observation of cell fixation and precise determination of critical drug concentrations.

Main Results:

• The assay successfully quantified the critical concentration of 2-deoxy-d-glucose to paralyze trypanosomes.
• Impacts on cell propulsion and energy generation were detected at significantly lower 2-deoxy-d-glucose concentrations.
• Suramin was shown to reduce trypanosome motility and induce cell lysis post-endocytosis.

Conclusions:

• The microfluidics-based assay enables rapid cell-based drug testing, distinguishing cytocidal from cytostatic effects.
• The method allows for screening effective drug dosages and investigating drug impacts on cell motility.
• This approach offers a powerful tool for advancing antiparasitic drug discovery and development.