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Related Experiment Videos

Single cell assay with an automated capillary microinjection system.

R Pepperkok1, C Schneider, L Philipson

  • 1European Molecular Biology, Laboratory, Heidelberg, Federal Republic of Germany.

Experimental Cell Research
|October 1, 1988
PubMed
Summary
This summary is machine-generated.

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This study introduces an automated microinjection system for high-throughput cell manipulation. The system offers precise control, high success rates for cytoplasmic and nuclear injections, and improved experimental accuracy.

Area of Science:

  • Cell biology
  • Biotechnology
  • Molecular biology

Background:

  • Manual microinjection techniques are time-consuming and prone to variability.
  • Accurate delivery of substances into cells is crucial for various biological studies.
  • Existing automated systems may lack precision or efficiency.

Purpose of the Study:

  • To describe an automated capillary microinjection system with computer-controlled positioning.
  • To evaluate the system's efficiency, reproducibility, and cell viability post-injection.
  • To demonstrate the system's utility in functional screening and biological discovery.

Main Methods:

  • Development of an automated system with computer-controlled cell and capillary positioning.
  • Optimization of glass capillary preparation and handling to reduce clogging.

Related Experiment Videos

  • Use of FITC-dextran as a coinjection marker to assess cell retrieval and viability.
  • Application of the system for mRNA studies and cDNA library screening.

    • Main Results:

    • High throughput: approximately 1500 injections per hour.
    • High success rates: >90% for cytoplasmic and >85% for nuclear injections.
    • Excellent cell viability: 99% retrieval 48 hours post-injection using FITC-dextran.
    • Improved accuracy in statistical evaluation compared to manual methods.

    Conclusions:

    • The automated microinjection system offers significant advantages in speed, reproducibility, and accuracy.
    • The system facilitates efficient functional screening of genetic material.
    • It enables confirmation of biological findings, such as the existence of DNA synthesis-inhibiting mRNA.