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

Luo，Yingdong1; Huang，Yuanyuan1; Li，Yani1; Duan，Xiudong1; Jiang，Yongguang2; Wang，Cong1; Fang，Jiakun3; Xi，Lei4; Nguyen，Nam Trung5; Song，Chaolong1

2023

10.1039/d3lc00127j

Lab on a Chip   影响因子和分区

1473-0197

1473-0189

Biomolecular imaging of intracellular structures of a single cell and subsequent screening of the cells are of high demand in metabolic engineering to develop strains with the desired phenotype. However, the capability of current methods is limited to population-scale identification of cell phenotyping. To address this challenge, we propose to utilize dispersive phase microscopy incorporated with a droplet-based microfluidic system that combines droplet volume-on-demand generation, biomolecular imaging, and droplet-on-demand sorting, to achieve high-throughput screening of cells with an identified phenotype. Particularly, cells are encapsulated in homogeneous environments with microfluidic droplet formation, and the biomolecule-induced dispersive phase can be investigated to indicate the biomass of a specific metabolite in a single cell. The retrieved biomass information consequently guides the on-chip droplet sorting unit to screen cells with the desired phenotype. To demonstrate the proof of concept, we showcase the method by promoting the evolution of the Haematococcus lacustris strain toward a high production of natural antioxidant astaxanthin. The validation of the proposed system with on-chip single-cell imaging and droplet manipulation functionalities reveals the high-throughput single-cell phenotyping and selection potential that applies to many other biofactory scenarios, such as biofuel production, critical quality attribute control in cell therapy, etc.

SCI ; EI

英语

通讯

National Natural Science Foundation of China["61804138","52177089","62022037"] ; Wuhan Science and Technology Bureau[2020010601012164] ; Fundamental Research Funds for the Central Universities, China University of Geosciences, Wuhan[CUG170608]

Biochemistry & Molecular Biology ; Chemistry ; Science & Technology - Other Topics ; Instruments & Instrumentation

Biochemical Research Methods ; Chemistry, Multidisciplinary ; Chemistry, Analytical ; Nanoscience & Nanotechnology ; Instruments & Instrumentation

WOS:000987196100001

ROYAL SOC CHEMISTRY

20232214171578

Drops ; Metabolic engineering ; Metabolism ; Metabolites ; Microfluidics ; Molecular biology ; Strain

Biomedical Engineering:461.1 ; Genetic Engineering:461.8.1 ; Biology:461.9 ; Microfluidics:632.5.1 ; Materials Science:951

CHEMISTRY

2-s2.0-85160422670

Scopus

1.A School of Mechanical Engineering and Electronic Information,China University of Geosciences,Wuhan,430074,China
2.School of Environmental Studies,China University of Geosciences,Wuhan,430074,China
3.State Key Laboratory of Advanced Electromagnetic Engineering and Technology,School of Electrical and Electronic Engineering,Huazhong University of Science and Technology,Wuhan,China
4.Department of Biomedical Engineering,Southern University of Science and Technology,Shenzhen,China
5.Queensland Micro and Nanotechnology Centre,Griffith University,Nathan,170 Kessels Road,QLD 4111,Australia

Luo，Yingdong,Huang，Yuanyuan,Li，Yani,et al. Dispersive phase microscopy incorporated with droplet-based microfluidics for biofactory-on-a-chip[J]. Lab on a Chip,2023,23(12):2766-2777.

Luo，Yingdong.,Huang，Yuanyuan.,Li，Yani.,Duan，Xiudong.,Jiang，Yongguang.,...&Song，Chaolong.(2023).Dispersive phase microscopy incorporated with droplet-based microfluidics for biofactory-on-a-chip.Lab on a Chip,23(12),2766-2777.

Luo，Yingdong,et al."Dispersive phase microscopy incorporated with droplet-based microfluidics for biofactory-on-a-chip".Lab on a Chip 23.12(2023):2766-2777.