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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics

| EXPLORING THE INTERFACE OF LIGHT AND BIOMEDICINE

  • Editor: Gregory W. Faris
  • Vol. 5, Iss. 7 — Apr. 26, 2010

Accommodating brightness and exposure levels in densitometry of stained polyacrylamide electrophoresis gels

Han Yen Tan, Tuck Wah Ng, and Oi Wah Liew  »View Author Affiliations


Applied Optics, Vol. 49, Issue 9, pp. 1623-1629 (2010)
http://dx.doi.org/10.1364/AO.49.001623


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Abstract

Flatbed scanner densitometers can be operated under various illumination and recording exposure levels. In this work, we show that optical density measurement accuracy, sensitivity, and stability of stained polyacrylamide electrophoresis gel densitometry are crucially dependent on these two factors (brightness and exposure level), notwithstanding that the source is monochromatic, spatially uniform, and the measurements are made using an accurately calibrated step wedge in tandem. We further outline a method to accommodate the intensity deviations over a range of illumination and exposure levels in order to maintain sensitivity and repeatability in the computed optical densities. Comparisons were also made with results from a commercial densitometer.

© 2010 Optical Society of America

OCIS Codes
(110.0110) Imaging systems : Imaging systems
(120.0120) Instrumentation, measurement, and metrology : Instrumentation, measurement, and metrology
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology

ToC Category:
Medical Optics and Biotechnology

History
Original Manuscript: November 9, 2009
Revised Manuscript: January 14, 2010
Manuscript Accepted: January 27, 2010
Published: March 15, 2010

Virtual Issues
Vol. 5, Iss. 7 Virtual Journal for Biomedical Optics

Citation
Han Yen Tan, Tuck Wah Ng, and Oi Wah Liew, "Accommodating brightness and exposure levels in densitometry of stained polyacrylamide electrophoresis gels," Appl. Opt. 49, 1623-1629 (2010)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=ao-49-9-1623


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References

  1. N. C. Kendrick, J. J. Johansen, P. R. Lee, and D. A. Santek, “Optimization of an HP Scanjet for quantification of protein electrophoresis gels,” Anal. Biochem. 219, 297-304 (1994). [CrossRef] [PubMed]
  2. S. G. Vincent, P. R. Cunningham, N. L. Stephens, A. J. Halayko, and J. T. Fisher, “Quantitative densitometry of proteins stained with Coomassie blue using a Hewlett Packard Scanjet scanner and Scanplot software,” Electrophoresis 18, 67-71 (1997). [CrossRef] [PubMed]
  3. H. Y. Tan, T. W. Ng, and O. W. Liew, “Light spectrum in flatbed scanner densitometry of stained polyacrylamide electrophoresis gels,” Biotech. 42, 474-478 (2007). [CrossRef]
  4. H. Y. Tan, T. W. Ng, and O. W. Liew, “Adapted LCD backlighting unit for densitometry of stained polyacrylamide electrophoresis gels,” Electrophoresis 30, 987-990 (2009). [CrossRef] [PubMed]
  5. H. Y. Tan and T. W. Ng, “Accurate step wedge calibration for densitometry of electrophoresis gels,” Opt. Commun. 281, 3013-3017 (2008). [CrossRef]
  6. J. Cabello, A. Bailey, I. Kitchen, M. Prydderch, A. Clark, R. Turchetta, and K. Wells, “Digital autoradiography using room temperature CCD and CMOS imaging technology,” Phys. Med. Biol. 52, 4993-5011 (2007). [CrossRef] [PubMed]
  7. A. Ferrero, J. Campos, and A. Pons, “Correction of photoresponse nonuniformity for matrix detectors based on prior compensation for their nonlinear behavior,” Appl. Opt. 45, 2422-2427 (2006). [CrossRef] [PubMed]
  8. M. Gassmann, B. Grenacher, B. Rohde, and J. Vogel, “Quantifying Western blots: pitfalls of densitometry,” Electrophoresis 30, 1845-1855 (2009). [CrossRef] [PubMed]
  9. U. K. Laemmli, “Cleavage of structural proteins during the assembly of the head of bacteriophage T4,” Nature 227, 680-685 (1970). [CrossRef] [PubMed]

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