January 1, 2013 Letizia Mancino HD MZ Localization Microscopy: “Molecular Galaxies 2013“ C. Cremer Institute of Molecular Biology (IMB), D-55128 MainzInstitute of Pharmacy and Molecular Biotechnology (IPMB)/Kirchhoff Institute of Physics (KIP), D-69120 University Heidelberg www.imb-mainz.de.
Molecular GalaxiesConventional Localization of Protein Transport Microscopy in a Human Blood- Brain Barrier Model O. Huber, A. Brunner, P. Maier, R. Kaufmann, P.-O. Couraud, C. Cremer, G. Fricker (2012)
Molecular Galaxies of Tight Junction Networks formed by individual Claudin Molecules in Human CellsConventional Microscopy Localization MicrocopyR. Kaufmann, J.Piontek, F. Grüll, M. Kirchgessner, J. Rossa, H.Wolburg, I. E. Blasig, C. Cremer (2012)
Molecular Galaxies of Gene TranscriptionConventional Microscopy Localization Microscopy Positions R. Kaufmann , C.Cremer, J.Gall (2012)
Deep Insights into the Interior of Molecular Galaxies: Single MoleculesSingle Molecules Nuclear Histones a) on Surface b) Inside Cell 300 nm Nucleus c),d) in Virus Single Viruses C. Cremer (2012)
Principle of Spectrally Assigned Localization Microscopy(SPDM, Spatial Position Determination Microscopy): OpticalIsolation of Diffraction Images of individual Molecules:Determination of Gravity Centers 3 point-like objects in x,y plane with next neigbour distances 50 nm a) Labelling with same spectral signature b) Labeling with different unique spectral signatures B,G,R Computation (scalar Theory): NA = 1.4, λexc = 488 nm Optical Isolation by the “Supernova“ Mode: Induction of stochastically distributed short flashes of light emission byCremer et al. 1996, 1999, 2010, 2012 individual molecules
Present State of Localization Microscopy (wide field)realized @ C. Cremer Lab (January 1, 2013)- Best Optical Resolution OR (resolvable distance): OR ~ 5 nm (~ 1/100 λexc, from localization precision)- Structural Resolution (imaging capability):● Mean (2D) distance between individual molecules actuallydetected: ~ 6 nm ( ~ 1/80 λexc)● Density of individually detected molecules : ~ 2,8 · 104/µm2● 3D single molecule resolution inside cells: ca. 20 nm laterally, 30 – 50 nm axially● Optical Sectioning: ~ 15 nm
Publications Cremer-Lab 2012 part IC. Cremer, Optics far Beyond the Diffraction Limit: From FocusedNanoscopy to Spectrally Assigned Localization Microscopy(2012). In: Springer Handbook of Lasers and Optics, 2nd edition(F. Träger, Edit.), pp. 1351 – 1389.T. Cremer, Y. Markaki, B. Hübner, A. Zunhammer, H. Strickfaden,S. Beichmanis, M. Heß, L. Schermelleh, M. Cremer, C. Cremer(2012) Chromosome Territory Organization within the Nucleus. In:Encyclopedia of Molecular Cell Biology and Molecular Medicine:Epigenetic Regulation and Epigenomics, 2nd Edition (R.A. Meyers,Edit.), pp. 1-30.Wiley-VCH.R. Kaufmann, C. Cremer, J. G. Gall (2012a) Superresolutionimaging of transcription units on newt lampbrush chromosomes.Chromosome Research, DOI 10.1007/s10577-012-9306-z-
Publications Cremer-Lab 2012 part IIR. Kaufmann, J. Piontek, F. Grüll, M. Kirchgessner, J.Rossa, H.Wolburg, I. E. Blasig, C. Cremer (2012b) Visualization andquantitative analysis of reconstituted tight junctions usinglocalization microscopy. PLoS One 7 (2 ) e31128: 1 – 9.O. Huber, A. Brunner, P. Maier, R. Kaufmann, P.-O. Couraud, C.Cremer, G. Fricker (2012) Localization microscopy (SPDM)reveals clustered formations of P-Glycoprotein in a human blood-brain barrier model. PLoS ONE 7 (9) e44776: 1-10.T. Ach, G. Best, S. Rossberger, R. Heintzmann, C. Cremer, S.Dithmar (2012) Autofluorescence imaging of human RPE cellgranules using structured illumination microscopy. Br. J.Ophthalmology, DOI 10.1136/bjophthalmol-2012-301547.