Nanoliter dispensing method by degassed poly(dimethylsiloxane) microchannels and its application in protein...

Area of Science:

• Biotechnology
• Materials Science
• Chemical Engineering

Background:

• Nanoliter-scale liquid handling is crucial for high-throughput screening.
• Existing methods for dispensing liquids into microwells can be complex and inefficient.

Purpose of the Study:

• To develop a simple and effective method for dispensing nanoliter volumes of liquid into microwell arrays.
• To demonstrate the utility of this method for protein crystallization screening and long-term incubation.

Main Methods:

• Utilized degassed poly(dimethylsiloxane) (PDMS) microchannels reversibly bound to microwell arrays.
• Employed PDMS as an internal vacuum source for liquid infusion.
• Tested compatibility with microwells made of PDMS, PMMA, and glass.

Main Results:

• Successfully dispensed aqueous solutions into microwells using the PDMS microchannel system.
• Achieved nanoliter-scale multiplex reactions and screening by forming microchambers.
• Demonstrated successful protein crystallization screening, including long-term incubation (over 2 months) in glass microwells.
• Obtained X-ray diffraction data with 3.1 Angstrom resolution from crystallized protein.

Conclusions:

• The developed method provides a facile approach for nanoliter liquid dispensing into microwell arrays.
• This technique is suitable for high-throughput screening, multiplex reactions, and protein crystallization.
• The system supports long-term incubation and yields crystals suitable for structural analysis.