Laborless, Automated Microfluidic Tandem Cell Processor for Visualizing Intracellular Molecules of Mammalian Cells

Visualization and quantification of intracellular molecules of mammalian cells are crucial steps in clinical diagnosis, drug development, and basic biological research. However, conventional methods rely mostly on labor-intensive, centrifugation-based manual operations for exchanging the cell carrie...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2020-02, Vol.92 (3), p.2580-2588
Main Authors: Mu, Tinglin, Toyoda, Hajime, Kimura, Yuki, Yamada, Masumi, Utoh, Rie, Umeno, Daisuke, Seki, Minoru
Format: Article
Language:English
Subjects:
Actin
Animals
Automation
Biological research
Cell Line
Cell Nucleus - chemistry
Cell Nucleus - metabolism
Centrifugation
Chemistry
Cytokines
Cytokines - analysis
Cytokines - biosynthesis
Cytoskeleton
Cytoskeleton - chemistry
Cytoskeleton - metabolism
Drug development
Equipment Design
Exchanging
Flow Cytometry - instrumentation
Interleukin 2
Intracellular
Lymphocytes
Lymphocytes T
Mammalian cells
Mammals
Mice
Microfluidic Analytical Techniques - instrumentation
Microfluidics
Microprocessors
NIH 3T3 Cells
Nuclei (cytology)
Pretreatment
Reaction time
Reagents
Recovery
Staining
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Summary:Visualization and quantification of intracellular molecules of mammalian cells are crucial steps in clinical diagnosis, drug development, and basic biological research. However, conventional methods rely mostly on labor-intensive, centrifugation-based manual operations for exchanging the cell carrier medium and have limited reproducibility and recovery efficiency. Here we present a microfluidic cell processor that can perform four-step exchange of carrier medium, simply by introducing a cell suspension and fluid reagents into the device. The reaction time period for each reaction step, including fixation, membrane permeabilization, and staining, was tunable in the range of 2 to 15 min by adjusting the volume of the reaction tube connecting the neighboring exchanger modules. We double-stained the cell nucleus and cytoskeleton (F-actin) using the presented device with an overall reaction period of ∼30 min, achieving a high recovery ratio and high staining efficiency. Additionally, intracellular cytokine (IL-2) was visualized for T cells to demonstrate the feasibility of the device as a pretreatment system for downstream flow-cytometric analysis. The presented approach would facilitate the development of laborless, automated microfluidic systems that integrate cell processing and analysis operations and would pave a new path to high-throughput biological experiments.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.9b04288