MPI Campus Seminar: Autonomous bioluminescence imaging of single cells

MPI Campus Seminar

  • Datum: 21.10.2020
  • Uhrzeit: 11:00 - 12:00
  • Vortragende(r): Carola Gregor
  • Department of NanoBiophotonics
  • Ort: Max-Planck-Institut für biophysikalische Chemie (MPIBPC)
  • Raum: Online
  • Gastgeber: S. Glöggler, A. Godec, A. Faesen, J. Liepe, S. Meek, A. Stein, M. Wilczek, S. Karpitschka, D. Zwicker, M. Oudelaar, L. Andreas
  • Kontakt: stefan.gloeggler@mpibpc.mpg.de
MPI Campus Seminar: Autonomous bioluminescence imaging of single cells
Bioluminescence is a process by which living cells can produce light. This phenomenon is realized in nature in many different ways which all follow the same principle of oxidation of a luciferin substrate by molecular oxygen, catalyzed by a luciferase enzyme. During this reaction, a product in an electronically excited state is formed, which releases the free energy of luciferin oxidation as a photon upon return to its ground state and thereby generates the bioluminescence light. The emission of bioluminescence can be used to image living cells and organisms without external light and hence without phototoxicity or bleaching, and also allows the observation of light-sensitive processes. The main problems of bioluminescence imaging are its low light levels and the need to externally supply the luciferin in most cases. However, no exogenous luciferin is required if the luciferin is synthesized and recycled by the cell itself. This can be achieved by coexpression of additional enzymes, as it was demonstrated with the bacterial bioluminescence system. Due to its comparatively low brightness, the bacterial bioluminescence system is currently not widely used. By several optimization procedures in different host cells, we have shown that the bacterial bioluminescence system can produce light levels that are comparable to firefly luciferase which requires an exogenous luciferin. This was achieved by mutagenesis of the involved enzymes and by improving their expression. The increased brightness enables long-term imaging of single bacterial and mammalian cells by bioluminescence microscopy without external substrates or light.
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