Quantitative Film Detection of 3H and 14C in Polyacrylamide Gels by Fluorography
European Journal of Biochemistry · 1975 · 4.7K citations · 8 references
Fluorography using PPO scintillator has been previously described, and recent revisions extend the method to agarose gels and include a rapid PPO recovery procedure. The study shows that briefly pre‑exposing film to light markedly boosts fluorography sensitivity. Pre‑exposure linearizes the film response, enabling quantitative micro‑densitometry of radioactivity. Micro‑densitometry of pre‑exposed film accurately reflects radioactivity distribution, detecting as low as 300 dis.
Methods which use the scintillator PPO to record film images of 3H in chromatograms and polyacrylamide gels (fluorography) have been described elsewhere. This paper demonstrates that pre-exposure of the film to a brief flash of light greatly increases the sensitivity of fluorography. Pre-exposure also permits quantitative interpretation of the film image, because it corrects the non-linear relationship between radioactivity of the sample and absorbance of the film image. Therefore the distribution of radioactivity in the sample is accurately represented by micro-densitometry of the image obtained on pre-exposed film. Using pre-exposed film 300 dis. 3H/min or 30 dis. 14C/min can be detected in a band in a gel in a 24-h exposure. The Appendix describes revisions and extensions of existing fluorographic procedures, including application to agarose gels and a rapid procedure for recovering PPO for re-use.
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A Film Detection Method for Tritium‐Labelled Proteins and Nucleic Acids in Polyacrylamide Gels
William M. Bonner, Ronald A. Laskey · European Journal of Biochemistry · 1974
8.7K citations
Use of Frog Eggs and Oocytes for the Study of Messenger RNA and its Translation in Living Cells
J. B. Gurdon, Charles D. Lane, Hugh R. Woodland et al. · Nature · 1971
774 citations
An evaluation of film detection methods for weak β-emitters, particularly tritium
Kurt Randerath · Analytical Biochemistry · 1970
277 citations
The theory of the photographic process
R.H. Oppermann · Journal of the Franklin Institute · 1943
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Low-Temperature Fluorography Induced by Tritium-Labelled Compounds on Thin-Layer Chromatograms
Ursula Lüthi, Peter G. Waser · Nature · 1965
65 citations