TY - JOUR
T1 - Two-dimensional, computer-controlled film scanner
T2 - Quantitation of fluorescence from ethidium bromide-stained DNA gels
AU - Sutherland, John Clark
AU - Monteleone, Denise C.
AU - Trunk, John
AU - Ciarrocchi, Giovanni
N1 - Funding Information:
We thank Dr. Anthony Blackett, Dr. Sharon Moore, Dr. Betsy Sutherland, and Dr. Gwyn Williams for helpful discussions and Dr. Alfred0 Luccio and Dr. Peter Takacs for the use of equipment. This work was supported by the United States Department of Energy.
PY - 1984/6
Y1 - 1984/6
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0021443661&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0021443661&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(84)90023-X
DO - 10.1016/0003-2697(84)90023-X
M3 - Article
C2 - 6206746
AN - SCOPUS:0021443661
SN - 0003-2697
VL - 139
SP - 390
EP - 399
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -