Injection Molded Microfluidics for Establishing High-Density Single Cell Arrays in an Open Hydrogel Format
| Autor: | Jeffrey D. Motschman, Sean T. Kelly, Benjamin B. Yellen, Ying Li |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Microfluidics
Injectable hydrogels High density HL-60 Cells Tumor cells Nanotechnology 010402 general chemistry 01 natural sciences Article Polyethylene Glycols Analytical Chemistry Automation Optical imaging Single-cell analysis Tumor Cells Cultured Humans Curing (chemistry) Chemistry Extramural Optical Imaging 010401 analytical chemistry technology industry and agriculture Hydrogels Microfluidic Analytical Techniques 0104 chemical sciences Acrylates Single-Cell Analysis K562 Cells |
| Zdroj: | Anal Chem |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.9b05099 |
| Popis: | Here, we develop an injection molded microfluidic approach for single cell analysis by making use of: 1) rapidly curing injectable hydrogels, 2) a high density microfluidic weir trap array, and 3) reversibly bonded PDMS lids that are strong enough to withstand the injection molding process, but which can be peeled off after the hydrogel sets. This approach allows for single cell patterns to be created with densities exceeding 40 cells per mm(2), is amenable to high speed imaging, and creates microfluidic devices that enable efficient nutrient and gas exchange and the delivery of specific biological and chemical reagents to individual cells. We show that it is possible to organize up to 10,000 single cells in a few minutes on the device, and we developed an image analysis program to automatically analyze the single-cell capture efficiency. The results show single cell trapping rates were better than 80%. We also demonstrate that the genomic DNA of the single cells trapped in the hydrogel can be amplified via localized, multiple displacement amplification in a massively parallel format, which offers a promising strategy for analyzing single cell genomes. Finally, we show the ability to perform selective staining of individual cells with a commercial bioprinter, providing proof of concept of its ability to deliver tailored reagents to specific cells in an array for future downstream analysis. This injection molded microfluidic approach leverages the benefits of both closed and open microfluidics, allows multi-day single cell cultures, direct access to the trapped cells for genotypic endpoint studies. |
| Databáze: | OpenAIRE |
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