Two-dimensional, computer-controlled film scanner: Quantitation of fluorescence from ethidium bromide-stained DNA gels

John Clark Sutherland, Denise C. Monteleone, John Trunk, Giovanni Ciarrocchi

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

A two-dimensional scanner based on a digital plotter is described. The device is used to analyze photographic negatives of ethidium bromide-stained DNA-agarose gels. Scanning is controlled by and photometric data transferred to a computer for processing, storage, display, and analysis such as integration of the areas under bands and determination of the mean distances of migration of polydisperse samples. An integral light source and detector module designed for reading optical "bar-codes" is mounted in place of the pen of the plotter. Spatial resolution and reproducibility are about 0.2 and 0.005 mm, respectively. Photometric precision as good as one part per thousand is achieved by sinusoidal modulation of the intensity of the light source and synchronous, phase-sensitive detection of the signal from the detector by a lock-in amplifier. No part of the sensor assembly touches the surface of the negative. In contrast to a densitometer, the computer transforms photometric data to values directly proportional to the amount of DNA at given points on the original gel. The ability to move the sensor in two dimensions over the negative allows for the integration across the width of a lane correctly allowing for the nonuniform distribution of the DNA.

Original languageEnglish (US)
Pages (from-to)390-399
Number of pages10
JournalAnalytical Biochemistry
Volume139
Issue number2
DOIs
StatePublished - Jun 1984
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Two-dimensional, computer-controlled film scanner: Quantitation of fluorescence from ethidium bromide-stained DNA gels'. Together they form a unique fingerprint.

  • Cite this