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A Multilayer Microfluidic Platform for the Conduction of Prolonged Cell-Free Gene Expression
Published on: October 6, 2019
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Systematic Analysis of Different Cell Spheroids with a Microfluidic Device Using Scanning Electrochemical Microscopy
Liang Zhao1, Mi Shi1, Yang Liu1
1Institute of Precision Medicine and Health, Research Center for Bioengineering and Sensing Technology, School of Chemistry and Biological Engineering, Beijing Key Laboratory for Bioengineering and Sensing Technology , University of Science and Technology Beijing , Beijing 100083 , China.
Analytical Chemistry
|March 15, 2019
Summary
This study introduces a novel microfluidic device for label-free imaging of 3D cell spheroids using scanning electrochemical microscopy (SECM). The technique enables simultaneous spheroid generation, electrochemical analysis, and gene expression profiling for tumorigenesis research.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Cell Biology
Background:
- 3D cell spheroids mimic in vivo conditions for tumorigenesis studies.
- Conventional optical imaging for spheroids requires fluorescent labels, potentially causing toxicity.
- Scanning electrochemical microscopy (SECM) offers label-free imaging but lacks systematic analysis capabilities for spheroids.
Purpose of the Study:
- To develop a microfluidic device for simultaneous generation and label-free SECM analysis of 3D cell spheroids.
- To enable systematic analysis of spheroid generation, electrochemical signals, and gene expression.
- To bridge SECM measurements with molecular biological analysis for heterotypic cell spheroids.
Main Methods:
- Development of a top-removable microfluidic device with micropit arrays for uniform spheroid production.
- Utilizing voltage-switching mode SECM with dual mediators for label-free alkaline phosphatase evaluation.
- Performing quantitative polymerase chain reaction (qPCR) assay on individual spheroids post-SECM imaging.
Main Results:
- Simultaneous production of uniformly sized breast tumor and fibroblast spheroids on a single device.
- Distinguished tumor aggregates from stroma by evaluating alkaline phosphatase activity using SECM.
- Enabled gene expression profiling on individual tumor or stromal spheroids.
Conclusions:
- The developed microfluidic device and SECM methodology provide a comprehensive platform for analyzing heterotypic 3D cell spheroids.
- This approach facilitates label-free, systematic analysis from spheroid generation to molecular profiling.
- The technique offers a valuable tool for tumorigenesis research by integrating electrochemical and molecular analyses.
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