Double Emulsion Picoreactors for High-Throughput Single-Cell Encapsulation and Phenotyping via FACS
| Autor: | Gaeun Kim, Kara Brower, Suzanne G. K. Calhoun, Lei S. Qi, Peter Suzuki, Margarita Khariton, Christopher J. Still, Bo Wang, Polly M. Fordyce |
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| Rok vydání: | 2020 |
| Předmět: |
Surface Properties
Cell Microfluidics Dispersity Double emulsion Article Analytical Chemistry Flow cytometry Cell Line Mice Single-cell analysis Mammalian cell High-Throughput Screening Assays Microscopy medicine Animals Particle Size Cell encapsulation medicine.diagnostic_test Chemistry Cell Encapsulation Cell sorting Microfluidic Analytical Techniques Flow Cytometry Cellular phenotype medicine.anatomical_structure Phenotype Biophysics Single-Cell Analysis |
| Zdroj: | Anal Chem |
| ISSN: | 1520-6882 |
| Popis: | In the past five years, droplet microfluidic techniques have unlocked new opportunities for the high-throughput genome-wide analysis of single cells, transforming our understanding of cellular diversity and function. However, the field lacks an accessible method to screen and sort droplets based on cellular phenotype upstream of genetic analysis, particularly for large and complex cells. To meet this need, we developed Dropception, a robust, easy-to-use workflow for precise single-cell encapsulation into picoliter-scale double emulsion droplets compatible with high-throughput phenotyping via fluorescence-activated cell sorting (FACS). We demonstrate the capabilities of this method by encapsulating five standardized mammalian cell lines of varying size and morphology as well as a heterogeneous cell mixture of a whole dissociated flatworm (5 - 25 μm in diameter) within highly monodisperse double emulsions (35 μm in diameter). We optimize for preferential encapsulation of single cells with extremely low multiple-cell loading events (ABSTRACT FIGURE |
| Databáze: | OpenAIRE |
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