Digital microfluidics for time-resolved cytotoxicity studies on single non-adherent yeast cells

Area of Science:

• Biotechnology
• Cell Biology
• Microfluidics

Background:

• Single cell analysis (SCA) is crucial for understanding cellular heterogeneity.
• Flow cytometry lacks spatio-temporal resolution for single cell studies.
• Non-adherent cells present challenges for high-throughput analysis.

Purpose of the Study:

• To introduce a digital microfluidic (DMF) platform for high-throughput SCA of non-adherent cells.
• To enable cytotoxicity assays with enhanced spatio-temporal resolution.
• To validate the DMF platform against bulk experimental results.

Main Methods:

• A DMF platform with a microwell array for trapping single non-adherent yeast cells.
• Shuttling cell-containing droplets over the microwell array.
• Time-lapse fluorescence microscopy to monitor cell responses to Amphotericin B over time.

Main Results:

• The DMF platform successfully performed high-throughput cytotoxicity assays on isolated yeast cells.
• Cellular responses were monitored with enhanced spatio-temporal resolution.
• DMF results showed strong correlation with bulk experimental data, validating the platform.

Conclusions:

• The developed DMF platform is a valuable tool for single cell analysis of non-adherent cells.
• The platform provides spatio-temporal resolution, crucial for understanding cellular behavior during treatment.
• The gentle cell trapping mechanism avoids cell damage, preserving cellular integrity.