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Improving Single-Cell Encapsulation Efficiency and Reliability through Neutral Buoyancy of Suspension.

ABSTRACT: Single-cell analysis is of critical importance in revealing cell-to-cell heterogeneity by characterizing individual cells and identifying minority sub-populations of interest. Droplet-based microfluidics has been widely used in the past decade to achieve high-throughput single-cell analysis. However, to maximize the proportion of single-cell emulsification is challenging due to cell sedimentation and aggregation. The purpose of this study was to investigate the influence of single-cell encapsulation and incubation through the use of neutral buoyancy. As a proof of concept, OptiPrep™ was used to create neutrally buoyant cell suspensions of THP-1, a human monocytic leukemia cell line, for single-cell encapsulation and incubation. We found that using a neutrally buoyant suspension greatly increased the efficiency of single-cell encapsulation in microdroplets and eliminated unnecessary cell loss. Moreover, the presence of OptiPrep™ was shown to not affect cellular viability. This method significantly improved the effectiveness of single-cell study in a non-toxic environment and is expected to broadly facilitate single-cell analysis.

SUBMITTER: Liu H

PROVIDER: S-EPMC7019761 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Improving Single-Cell Encapsulation Efficiency and Reliability through Neutral Buoyancy of Suspension.

Liu Hangrui H Li Ming M Wang Yan Y Piper Jim J Jiang Lianmei L

Micromachines 20200115 1

Single-cell analysis is of critical importance in revealing cell-to-cell heterogeneity by characterizing individual cells and identifying minority sub-populations of interest. Droplet-based microfluidics has been widely used in the past decade to achieve high-throughput single-cell analysis. However, to maximize the proportion of single-cell emulsification is challenging due to cell sedimentation and aggregation. The purpose of this study was to investigate the influence of single-cell encapsula ...[more]

PMID: 31952228

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Project description:Introduction"Lean" is a set of management principles which focus on increasing value and efficiency by reducing or avoiding waste (e.g., overproduction, defects, inventory, transportation, waiting, motion, over processing). It has been applied to manufacturing, education, and health care, leading to optimized process flow, increased efficiency and increased team empowerment. However, to date, it has not been applied to neuroimaging research.MethodsLean principles, such as Value stream mapping (e.g. a tool with which steps in the workflow can be identified on which to focus improvement efforts), 5S (e.g. an organizational method to boost workplace efficiency and efficacy) and Root-cause analysis (e.g. a problem-solving approach to identify key points of failure in a system) were applied to an ongoing, large neuroimaging study that included seven research visits per participant. All team members participated in a half-day exercise in which the entire project flow was visualized and areas of inefficiency were identified. Changes focused on removing obstacles, standardization, optimal arrangement of equipment and root-cause-analysis. A process for continuous improvement was also implemented. Total time of an experiment was recorded before implementation of Lean for two participants and after implementation of Lean for two participants. Satisfaction of team members was assessed anonymously on a 5-item Likert scale, ranging from much worsened to much improved.ResultsAll team members (N = 6) considered the overall experience of conducting an experiment much improved after implementation of Lean. Five out of six team members indicated a much-improved reduction in time, with the final team member considering this somewhat improved. Average experiment time was reduced by 13% after implementation of Lean (from 142 and 147 minutes to 124 and 128 minutes).DiscussionLean principles can be successfully applied to neuroimaging research. Training in Lean principles for junior research scientists is recommended.

Dataset Information

Improving Single-Cell Encapsulation Efficiency and Reliability through Neutral Buoyancy of Suspension.

Publications

Improving Single-Cell Encapsulation Efficiency and Reliability through Neutral Buoyancy of Suspension.

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