TechRxiv
Conchouso_et_al-DropletMicrofluidicTools.pdf (15.09 MB)
Download file

Integration of Droplet Microfluidic Tools for Single-cell Functional Metagenomics: An Engineering Head Start

Download (15.09 MB)
preprint
posted on 11.06.2021, 05:05 by David Conchouso, Amani Al-Ma’abadi, Hayedeh Behzad, Mohammed Alarawi, Masahito Hosokawa, Yohei Nishikawa, Haruko Takeyama, Katsuhiko MinetaKatsuhiko Mineta, Takashi Gojobori

Droplet microfluidics techniques have shown promising results to study single-cells at high throughput. However, their adoption in laboratories studying “-omics” sciences is still irrelevant because of the field’s complex and multidisciplinary nature. To facilitate their use, here we provide engineering details and organized protocols for integrating three droplet-based microfluidic technologies into the metagenomic pipeline to enable functional screening of bioproducts at high throughput. First, a device encapsulating single-cells in droplets at a rate of ~ 250 Hz is described considering droplet size and cell growth. Then, we expand on previously reported fluorescent activated droplet sorting (FADS) systems to integrate the use of 4 independent fluorescence-exciting lasers (e.g., 405, 488, 561, 637 nm) in a single platform to make it compatible with different fluorescence-emitting biosensors. For this sorter, both hardware and software are provided and optimized for effortlessly sorting droplets at 60 Hz. Then, a passive droplet merger was also integrated into our method to enable adding new reagents to already made droplets at a rate of 200 Hz. Finally, we provide an optimized recipe for manufacturing these chips using silicon dry-etching tools. Because of the overall integration and the technical details presented here, our approach allows biologists to quickly use microfluidic technologies and achieve both single-cell resolution and high-throughput (> 50,000 cells/day) capabilities to mining and bioprospecting metagenomic data.

History

Email Address of Submitting Author

katsuhiko.mineta@kaust.edu.sa

ORCID of Submitting Author

0000-0002-4727-045X

Submitting Author's Institution

King Abdullah University of Science and Technology

Submitting Author's Country

Saudi Arabia

Usage metrics

Categories

Exports