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Related Experiment Videos

The LightCycler: a microvolume multisample fluorimeter with rapid temperature control

C T Wittwer1, K M Ririe, R V Andrew

  • 1Idaho Technology, Idaho Falls, USA. ctwittwer@msscc.med.utah.edu

Biotechniques
|January 1, 1997
PubMed
Summary
This summary is machine-generated.

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New microvolume fluorimeters offer rapid temperature control for analyzing small samples. These instruments enable fast, precise quantification and nucleic acid analysis, including DNA amplification in 10-15 minutes.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Microvolume fluorimeters are essential for analyzing limited or precious samples.
  • Rapid temperature control is crucial for kinetic assays and nucleic acid analysis.
  • Existing technologies may lack the speed and sensitivity required for certain applications.

Purpose of the Study:

  • To describe experimental and commercial microvolume fluorimeters with rapid temperature control.
  • To showcase the application of these instruments in various analytical techniques.
  • To demonstrate the efficiency of these systems for rapid DNA amplification and analysis.

Main Methods:

  • Utilized fluorescence optics from flow cytometry to analyze 1-10 microL samples in glass capillaries.

Related Experiment Videos

  • Implemented a circulating air vortex for homogeneous temperature control and rapid sample temperature changes (10°C/s).
  • Developed prototype and utilized commercial instruments (e.g., LightCycler) with different excitation sources and detection systems.

    • Main Results:

    • Achieved rapid temperature cycling (10°C/s) and precise sample interrogation in microvolumes.
    • Demonstrated successful application in analyte quantification, nucleic acid melting curves, enzyme assays, and fluorescence resonance energy transfer.
    • Successfully monitored rapid cycle DNA amplification using three distinct fluorescence techniques within 10-15 minutes.

    Conclusions:

    • Microvolume fluorimeters with rapid temperature control provide a powerful tool for high-throughput and sensitive analyses.
    • These instruments significantly reduce analysis time, particularly for applications like DNA amplification.
    • The technology enables efficient analysis of small or expensive samples, advancing molecular diagnostics and research.