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Title: High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures

Abstract

Microbiomes exert significant influence on our planet's ecology. Elucidating the identities of individual microbes within these communities and how they interact is a vital research imperative. Using traditional plating and culturing methods, it is impractical to assess even a small fraction of the interactions that exist within microbial communities. To address this technology gap, we integrated gel microdroplet technology with microfluidics to generate millions of microdroplet cultures (MDs) that sequester individual cells for phenotyping MDs, facilitating rapid analysis and viable recovery using flow cytometry. Herein, we describe a validated high-throughput phenotyping pipeline that elucidates cell-to-cell interactions for millions of combinations of microorganisms. Through iterative co-culturing of an algae and a pool of environmentally sourced microbes, we successfully isolated bacteria that improved algal growth.

Authors:
 [1];  [2]; ORCiD logo [2];  [3]; ORCiD logo [2];  [2]; ORCiD logo [2]; ORCiD logo [4]
  1. Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Oregon State Univ., Corvallis, OR (United States)
  2. Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
  3. (Tony) [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
  4. Los Alamos National Lab. (LANL), Los Alamos, NM (United States); New Mexico Consortium, Los Alamos, NM (United States)
Publication Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org.:
USDOE Laboratory Directed Research and Development (LDRD) Program
OSTI Identifier:
1532721
Report Number(s):
LA-UR-19-21407
Journal ID: ISSN 0736-6205
Grant/Contract Number:  
89233218CNA000001
Resource Type:
Accepted Manuscript
Journal Name:
Biotechniques
Additional Journal Information:
Journal Volume: 66; Journal Issue: 5; Journal ID: ISSN 0736-6205
Country of Publication:
United States
Language:
English
Subject:
microfluidics; high-throughput screening; microdroplet; picoculture; flow cytometry; algae; emulsion; encapsulation

Citation Formats

Ohan, Juliette Asdegegh, Pelle, Benjamin, Nath, Pulak, Huang, Jen -Huang, Hovde, Blake, Vuyisich, Momchilo, Dichosa, Armand Earl Ko, and Starkenburg, Shawn Robert. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures. United States: N. p., 2019. Web. doi:10.2144/btn-2018-0124.
Ohan, Juliette Asdegegh, Pelle, Benjamin, Nath, Pulak, Huang, Jen -Huang, Hovde, Blake, Vuyisich, Momchilo, Dichosa, Armand Earl Ko, & Starkenburg, Shawn Robert. High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures. United States. doi:10.2144/btn-2018-0124.
Ohan, Juliette Asdegegh, Pelle, Benjamin, Nath, Pulak, Huang, Jen -Huang, Hovde, Blake, Vuyisich, Momchilo, Dichosa, Armand Earl Ko, and Starkenburg, Shawn Robert. Fri . "High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures". United States. doi:10.2144/btn-2018-0124. https://www.osti.gov/servlets/purl/1532721.
@article{osti_1532721,
title = {High-throughput phenotyping of cell-to-cell interactions in gel microdroplet pico-cultures},
author = {Ohan, Juliette Asdegegh and Pelle, Benjamin and Nath, Pulak and Huang, Jen -Huang and Hovde, Blake and Vuyisich, Momchilo and Dichosa, Armand Earl Ko and Starkenburg, Shawn Robert},
abstractNote = {Microbiomes exert significant influence on our planet's ecology. Elucidating the identities of individual microbes within these communities and how they interact is a vital research imperative. Using traditional plating and culturing methods, it is impractical to assess even a small fraction of the interactions that exist within microbial communities. To address this technology gap, we integrated gel microdroplet technology with microfluidics to generate millions of microdroplet cultures (MDs) that sequester individual cells for phenotyping MDs, facilitating rapid analysis and viable recovery using flow cytometry. Herein, we describe a validated high-throughput phenotyping pipeline that elucidates cell-to-cell interactions for millions of combinations of microorganisms. Through iterative co-culturing of an algae and a pool of environmentally sourced microbes, we successfully isolated bacteria that improved algal growth.},
doi = {10.2144/btn-2018-0124},
journal = {Biotechniques},
number = 5,
volume = 66,
place = {United States},
year = {2019},
month = {5}
}

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