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High performance gel imaging with a commercial single lens reflex camera.

J Slobodan1, R Corbett, N Wye

  • 1Canada's Michael Smith Genome Sciences Centre, BC Cancer Agency, Suite 100 570 W 7th Ave, Vancouver, BC V5Z 4S6, Canada.

The Review of Scientific Instruments
|April 5, 2011
PubMed
Summary

A cost-effective gel imaging system using a DSLR camera matches laser scanner performance for high-throughput DNA fingerprinting. This system offers comparable specificity and sensitivity for analyzing DNA bands with SYBR Green.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Imaging Technology

Background:

  • High-throughput DNA fingerprinting is crucial for genetic analysis.
  • Existing laser scanners offer high performance but can be expensive.
  • There is a need for affordable, high-performance gel imaging solutions.

Purpose of the Study:

  • To construct and evaluate a high-performance gel imaging system using a digital single-lens reflex (DSLR) camera.
  • To compare its performance against a laser scanner for DNA fingerprinting applications.
  • To assess the impact of different color filters on imaging quality.

Main Methods:

  • A gel imaging system was built using a DSLR camera and epi-illumination.
  • The system was used to image large agarose gels (19 × 23 cm) with numerous DNA bands.

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  • Performance was evaluated by comparing specificity and sensitivity with a laser scanner using SYBR Green.
  • Various low and high-cost color filters were tested.

    • Main Results:

    • The DSLR-based imager demonstrated equivalent performance to a laser scanner for high-throughput DNA fingerprinting.
    • Specificity and sensitivity were within 1% of the laser scanner when using identical band identification software.
    • Inexpensive dyed acrylic filters, when carefully combined with dielectric interference filters, yielded good results.
    • Poor filter combinations were also identified, highlighting the importance of selection.

    Conclusions:

    • A DSLR camera-based gel imaging system provides a cost-effective alternative to laser scanners for DNA fingerprinting.
    • Careful selection of optical filters is essential for achieving optimal performance.
    • The study proposes methods for evaluating imager resolution, dynamic range, and optical efficiency to aid system comparison.