Nystatin-induced changes in yeast monitored by time-resolved automated single cell electrorotation

Area of Science:

• Biophysics
• Cell Biology
• Analytical Chemistry

Background:

• Electrorotation is valuable for determining cellular properties but limited by manual data collection.
• Automated systems are needed to overcome limitations in real-time data acquisition for electrorotation.

Purpose of the Study:

• To develop an automated system for real-time electrorotation spectra collection of single cells.
• To monitor cellular changes in yeast Saccharomyces cerevisiae exposed to the antibiotic nystatin.

Main Methods:

• A novel system utilizing hardware-based image moment registration for automatic object orientation calculation.
• Real-time video signal processing without intermediate image storage, achieving high data throughput (approx. 2 recordings/sec).
• Electrorotation spectra collected from single yeast cells in the 1 kHz to 1 GHz frequency range.

Main Results:

• The system successfully collected electrorotation spectra automatically and in real-time.
• Changes in yeast cell membrane permeability and cytoplasmic conductivity were observed after nystatin addition.
• Two distinct events at 7 and 75 minutes post-nystatin exposure showed rapid increases in membrane permeability and decreased cytoplasmic conductivity.

Conclusions:

• The developed automated system significantly enhances the utility of electrorotation for studying dynamic cellular processes.
• Real-time monitoring reveals rapid alterations in yeast cell membrane and cytoplasm properties under antibiotic stress.
• This technology offers high temporal resolution for investigating drug-induced cellular responses.