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Related Experiment Videos

Optimization of reporter cells for expression profiling in a microfluidic device.

Kenneth J Wieder1, Kevin R King, Deanna M Thompson

  • 1Center for Engineering in Medicine/Department of Surgery, Massachusetts General Hospital, Shriners Hospitals for Children, Harvard Medical School, Boston, MA 02114, USA.

Biomedical Microdevices
|September 1, 2005
PubMed
Summary
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Green fluorescence protein (GFP) technologies enable cell monitoring. A microfluidic Living Cell Array (LCA) optimizes GFP reporter cell characteristics for high-throughput gene expression profiling.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Cell Biology

Background:

  • Green fluorescence protein (GFP) technologies allow non-invasive monitoring of cellular functions and gene expression.
  • Traditional methods using GFP are limited in scalability, high-throughput analysis, and single-cell measurements.
  • Microfluidic devices offer potential for advanced cellular analysis.

Purpose of the Study:

  • To optimize GFP reporter cell characteristics within a microfluidic Living Cell Array (LCA) for enhanced gene expression profiling.
  • To evaluate the performance of GFP reporter cells in the LCA for dynamic expression analysis.
  • To establish criteria for utilizing reporter cell lines in microfluidic devices for live cell expression profiling.

Main Methods:

  • Development of a microfluidic Living Cell Array (LCA) for high-throughput GFP expression monitoring.

Related Experiment Videos

  • Generation and utilization of a reporter cell line for the transcription factor NF-kappa B.
  • Stimulation of reporter cells with TNF-alpha within the LCA to monitor fluorescence kinetics.
  • Optimization through reporter cell line subcloning and cell cycle synchronization.

    • Main Results:

    • GFP reporter cells in the LCA exhibit fluorescence kinetics comparable to standard tissue culture upon TNF-alpha stimulation.
    • The LCA successfully monitors multiple expression waves from small cell populations.
    • Subcloning and cell cycle synchronization enhance fluorescence readout, improving dynamic expression profiling.
    • Established criteria for reporter cell line optimization in microfluidic devices.

    Conclusions:

    • The Living Cell Array (LCA) is a viable platform for high-throughput, dynamic gene expression profiling using GFP reporter cells.
    • Optimization of reporter cell characteristics is crucial for maximizing fluorescence readout and improving microfluidic-based live cell analysis.
    • This study provides essential guidelines for employing reporter cell lines in microfluidic systems for advanced cellular studies.