An integrative temperature-controlled microfluidic system for budding yeast heat shock response analysis at the...

Area of Science:

• Cellular biology
• Biophysics
• Microfluidics

Background:

• Budding yeast is a key model organism for studying cellular stress responses.
• Precise environmental control, especially temperature, is crucial for high-resolution single-cell studies.
• Existing platforms lack the capability for fine-tuned temperature control and diverse thermal change patterns.

Purpose of the Study:

• To develop a novel, low-cost, and easily fabricated microfluidic platform for precise temperature control in yeast.
• To enable the study of cellular responses to various temperature change dynamics (step, gradient, oscillations).
• To investigate the dynamic behavior of proteins, such as transcription factor Msn2, under controlled thermal stress.

Main Methods:

• Development of a microfluidic chip integrating a liquid metal microheater and a liquid metal vs. conductive PDMS thermocouple.
• Automated equipment for generating three distinct temperature change profiles: step, gradient, and periodical oscillations.
• Observation of transcription factor Msn2's nuclear entry and exit in yeast cells subjected to heat stress (37 °C).

Main Results:

• Successful fabrication of a cost-effective and simple-to-produce temperature-controlled microfluidic platform.
• Demonstration of precise control over step, gradient, and oscillating temperature changes.
• Validation of the platform's utility by observing Msn2 protein dynamics in response to controlled heat stress.

Conclusions:

• The developed microfluidic platform offers a powerful tool for single-cell analysis of cellular responses to dynamic thermal environments.
• It overcomes limitations of previous systems, enabling detailed studies of cellular adaptation strategies.
• The platform is feasible for investigating dynamic protein behaviors in yeast under precisely controlled temperature conditions.