A microfluidic flow-focusing device for low sample consumption serial synchrotron crystallography experiments in...

Area of Science:

• Structural Biology
• Biophysics
• Materials Science

Background:

• Serial synchrotron crystallography (SSC) requires specialized sample environments for low-dose, room-temperature protein structure determination.
• Efficient protein production and crystallization are critical, necessitating X-ray-compatible setups that minimize sample usage and maintain native crystal conditions.

Purpose of the Study:

• To present an optimized and rapid manufacturing process for metal-polyimide microfluidic flow-focusing devices.
• To enable X-ray diffraction data collection from protein microcrystals in a continuous liquid flow.

Main Methods:

• Fabrication of metal-polyimide microfluidic chips using a fast and optimized manufacturing route.
• Implementation of flow-focusing conditions within the microfluidic devices for sample delivery.
• Utilizing high-repetition-rate X-ray sources for data collection.

Main Results:

• The developed microfluidic devices facilitate X-ray diffraction data collection in a flow format.
• Flow-focusing significantly reduces sample consumption.
• Enables complex experiments like rapid mixing for time-resolved serial crystallography.
• Full datasets can be acquired rapidly (approximately 1 hour) from crystal slurries in liquid flow.
• The microfluidic chips are manufacturable, reliable, durable, and operate at low flow rates (∼30 µl/h).

Conclusions:

• The novel microfluidic devices offer an efficient and low-sample-consumption platform for serial synchrotron crystallography.
• This technology accelerates protein structure determination and opens possibilities for advanced time-resolved experiments.
• The manufacturability and performance of the chips make them suitable for routine structural biology applications.