Printing Life-Inspired Subcellular Scale Compartments with Autonomous Molecularly Crowded Confinement

Area of Science:

• Biotechnology
• Chemical Engineering
• Molecular Biology

Background:

• Preparing life-inspired subcellular compartments is challenging.
• Understanding molecular behavior in confined environments is crucial for cell biology.

Purpose of the Study:

• To develop a simple, rapid, and controlled platform for creating life-inspired subcellular scale compartments.
• To investigate molecular behavior and interactions within these engineered compartments.

Main Methods:

• Inkjet printing of femtoliter (fL)-scale aqueous droplets with programmed molecular content.
• Utilizing nanoliter (nL) mineral oil drop arrays for droplet containment.
• Employing fluorescence microscopy and fluorescence lifetime imaging for analysis.
• Studying molecular probes, DNA hairpins, enzymatic cascades, and protein-ligand interactions.

Main Results:

• The platform successfully generated stable fL-droplets for several hours.
• Submicrometer, molecularly crowded shell structures formed at droplet surfaces.
• Specific features like heterogeneity and responsivity to molecular triggers were observed.
• Intermolecular interactions within the confined environments were elucidated.

Conclusions:

• The developed inkjet printing platform provides a novel method for creating nature-inspired confined reactors.
• This technology facilitates a deeper understanding of molecular confinement effects relevant to subcellular compartments.
• The platform holds potential for advanced lab-on-chip studies and biomolecular research.