Dispersive phase microscopy incorporated with droplet-based microfluidics for biofactory-on-a-chip

Area of Science:

• Biotechnology and Metabolic Engineering
• Microfluidics and Single-Cell Analysis

Background:

• Current cell phenotyping methods are limited to population-scale analysis, hindering precise strain development.
• Metabolic engineering requires high-throughput screening of single cells for desired phenotypes.

Purpose of the Study:

• To develop a high-throughput single-cell phenotyping and screening system.
• To enable precise identification and selection of cells based on intracellular biomolecule content.

Main Methods:

• Integration of dispersive phase microscopy with droplet-based microfluidics.
• Utilizing droplet volume-on-demand generation, biomolecular imaging, and droplet-on-demand sorting.
• Encapsulating single cells in homogeneous microfluidic droplets for analysis.

Main Results:

• Demonstrated high-throughput single-cell imaging and phenotype-based sorting.
• Successfully applied the system to evolve *Haematococcus lacustris* for enhanced astaxanthin production.
• Validated the system's potential for single-cell biomass quantification and selection.

Conclusions:

• The proposed microfluidic system offers a powerful tool for high-throughput single-cell phenotyping and selection.
• This technology has broad applicability in metabolic engineering, biofuel production, and cell therapy quality control.
• Enables precise screening of cells based on intracellular metabolite biomass.