Three-dimensional imaging of DNA fragments during electrophoresis using a confocal detector

Laurence R. Brewer; J. Courtney Davidson; Joseph W. Balch; Anthony Carrano

doi:10.1117/12.206035

3 April 1995 Three-dimensional imaging of DNA fragments during electrophoresis using a confocal detector

Laurence R. Brewer, J. Courtney Davidson, Joseph W. Balch, Anthony Carrano

Author Affiliations +

Proceedings Volume 2386, Ultrasensitive Instrumentation for DNA Sequencing and Biochemical Diagnostics; (1995) https://doi.org/10.1117/12.206035
Event: Photonics West '95, 1995, San Jose, CA, United States

Abstract

We have measured the 3D distribution of DNA fragments within an electrophoretic band. The measurements were made using a confocal microscope and a photon counting photomultiplier detector. A DNA sequencing standard was loaded into glass microchannel plates containing polyacrylamide gel. The measurements were made by scanning the plates in three dimensions using a mechanical stage under computer control, while electrophoresis was taking place. We found that the distribution of DNA was the same for all the bands measured in the sequencing ladder with an approximate Gaussian distribution along all three axis. These measurements are important to understand what physical forces shape electrophoretic bands confined by a channel and also as an aid in the design of high throughput DNA sequencers.

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Laurence R. Brewer, J. Courtney Davidson, Joseph W. Balch, and Anthony Carrano "Three-dimensional imaging of DNA fragments during electrophoresis using a confocal detector", Proc. SPIE 2386, Ultrasensitive Instrumentation for DNA Sequencing and Biochemical Diagnostics, (3 April 1995); https://doi.org/10.1117/12.206035

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