Abstract
Digital imaging spectroscopy has been used to obtain the grayscale spectrum of colored bacterial colonies directly from petri dishes. Up to 500 individual colony spectra can be simultaneously recorded and processed from a single plate. Spectra can be obtained in the visible to near infrared region (400nm–900nm) with lOnm resolution. Instrument response is normalized through run-time radiometric calibration such that each grayscale spectrum can be converted to the ground-state absorption spectrum of the colony. In this study, mutants of the photosynthetic bacterium Rhodobacter capsulatus have been differentiated by the absorption spectra of their pigment-protein complexes. This imaging technique is applicable to chro mogenic systems in which colony and/or media color (e.g. indicator plates) provides a quantitative indicator of gene expression.
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References
Yang, M.M., Youvan D.C. 1988. Applications of imaging spectroscopy in molecular biology: I. Screening photosynthetic bacteria. Bio/Technology, 6: 939–942.
Youvan, D.C., Ismail, S., Bylina, E.J. 1985. Chromosomal deletion and plasmid complementation of the photosynthetic reaction center and light-harvesting genes from Rhodopseudomonas capsulata. Gene 38: 19–30.
Amesz, J. 1978. Fluorescence and energy transfer, p. 333–340. In: The Photosynthetic Bacteria. R. K. Clayton and W. R. Sistrom. (Ed.). Plenum Press, New York.
Youvan, D.C., Hearst, J.E., Marrs, B.L. 1983. Isolation and characterization of enhanced fluorescence mutants of Rhodobacter capsulata. J. of Bacteriology. 154: 748–755.
Demtroder, W. 1988. Laser Spectroscopy: Basic Concepts and Instrumentation, p. 160–168. 3rd printing. Springer-Verlag, NY.
Bylina, E.J., Robles, S.J., Youvan, D.C. 1988. Directed mutations affecting the putative bacteriochlorophyll-binding sites in the light harvesting antenna I of Rhodobacter capsulatus. Israel J. of Chemistry. 28: 73–78.
Latimer, P., Eubanks C.A.H. 1962. Absorption spectrophotometry of turbid suspensions: A method of correcting for large systematic distortions. Archives of Biochemistry and Biophysics. 98: 274–285.
Duysens, L.N.M. 1956. The flattening of the absorption spectrum of suspensions, as compared to that of solutions. Biochimica et Biophysica Acta. 19: 1–12.
Breton, J., Nabedryk, E. 1987. Pigment and protein organization in reaction center and antenna complexes, p. 159–195. In: The Light Reactions. J. Barber (Ed.). Elsevier Science Publishers B.V., New York.
Kasha, M. 1963. Energy transfer mechanisms and the molecular exciton model for molecular aggregates. Radiation Research. 20: 55–71.
Miller,J.H. 1972. Experiments in Molecular Genetics, p. 47–55. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Reidhaar-Olson,J.F., Sauer, R.T. 1988. Combinatorial cassette mutagenesis as a probe of the informational content of protein sequences. Science 241: 53–57.
Bird, R.E., Hardman, K.D., Jacobson, J.W., Johnson, S., Kaufman, B.M., Lee, S., Lee, T., Pope, S.H., Riordan, G.S., Whitlow, M. 1988. Single-chain antigen-binding proteins. Science 242: 423–426.
Matayoshi, E.D., Wang, G.T., Krafft, G.A., Erickson, J. 1990. Novel fluorogenic substrates for assaying retroviral proteases by resonance energy transfer. Science 247: 954–247.
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Arkin, A., Goldman, E., Robles, S. et al. Applications of Imaging Spectroscopy in Molecular Biology II. Colony Screening Based on Absorption Spectra. Nat Biotechnol 8, 746–749 (1990). https://doi.org/10.1038/nbt0890-746
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DOI: https://doi.org/10.1038/nbt0890-746
