S1007 Doing structural biology with the electron microscope C9940 3-Dimensional Transmission electron microscopy Lecture 1: Introduction & history 2 Course organization  Time and place: Monday 14:00 – 15:50, A35/211  Language: English  Finishing the course: exam (written form), 4 credits  Jiri Novacek (jiri.novacek@ceitec.muni.cz) 3 Syllabus 12th March History 19th March No lecture 26th March Anatomy of electron microscope 2nd April Easter holiday (no lecture) 9th April Specimen preparation 16th April Principles of image formation 23rd April Tomography 30th April 3D volume generation 7th May Single particle analysis 14th May Atomic modeling and validation 21st May Recent advances 4 Resources Literature: – J. Frank, Three-dimensional electron microscopy of macromolecular assembliesvisualization of biological molecules in their native state. – J. Frank, Electron Tomography: Methods for Three-Dimensional Visualization of Structures in the Cell – Williams et al., Transmission electron microscopy Video courses (youtube): – Grant Jensen – NRAMM SEMC – Cryo-EM14 (LMB) – Cryo-EM17 (LMB) 5 History of electron microscopy 6 Munich: Ernst Ruska & Otto Scherzer Ernst (and Helmut) Ruska: timeline 1930 1940 1950 1960 1970 1980 1990 2000 2010 Milestones: 1931: Invention of the electrostatic lens magnification: 400X 1933: First electron microscope mag: 7000X (vs. LM: 2000X) 1933: Completed Ph.D. (!) 1939: First (viable) commercial EM mag: 100,000X 1939: His brother Helmut Ruska images first virus (TMV) 1952: Helmut moves from Siemens to Albany 1986: Nobel Prize in Physics Ernst Ruska http://www.biografiasyvidas.com http://ernst.ruska.de Sketch from 1929 9 Ernst Ruska Replica of first electron microscope First (viable) commercial microscope (Siemens) http://www.bluesci.org http://ernst.ruska.de http://ernst.ruska.de First Siemens microscope, 1939 First commercial EM (1937) was Metropolitan-Vickers EM1 (EM2 shown) The first commercial electron microscope was actually by the British company Metropolitan-Vickers in 1937. However, the magnification was worse than for the light microscope, so the Siemens is considered “first.” http://emu.msim.org.uk Clarification Otto Scherzer: timeline 1930 1940 1950 1960 1970 1980 1990 2000 2010 Milestones: 1934: First comprehensive book on electron optics 1936: Spherical aberration → resolution 50-100X the wavelength 1947-1951: Devised correction schemes for aberration correction 1949: “Can atoms be visible in the electron microscope?” “Scherzer focus” Scherzer → Harald Rose (Wadsworth, Darmstadt) → Max Haider Otto Scherzer http://www.microscopy.org First book on electron optics http://www.microscopy.org Scherzer (1949) Physikalische Blätter & Scherzer (1949) Journal of Applied Physics http://www.microscopy.org http://www.microscopy.org “Can atoms be visible in the electron microscope?” “Scherzer focus” Toronto group: E.F. Burton, James Hillier, etc. James Hillier: timeline 1930 1940 1950 1960 1970 1980 1990 2000 2010 Milestones: 1940 February Hillier started at RCA enlisted by Vladimir Dworykin (cathode ray tube) 1940 Jul 4: Commercial EM, Model B (EMB) 1941: 300kV, for dealing with thick specimens 1947: first stigmator stigmators were iron screws tapped into the pole piece resolution → 1nm 1973 (as VP of RCA): first videodisc James Hillier Seated, with Albert Prebus standing At RCA Model B, 1940 Electron microscopy in the Czech Republic 1930 1940 1950 1960 1970 1980 1990 2000 2010 Milestones: 1950: Ales Blaha: “Tripod” at the Institute of Theoretical and Experimentation Electrotechnology of the Technical University 1951: Tesla BS241 first Czech commercial microscope 50kV resolution: 2nm 1968: First ultrahigh vacuum system (Institute of Scientific Instruments) 1990: Delong Instruments founded 1991: TESCAN founded by engineers from Tesla (TEsla SCANing) 1997: FEI builds factory in Brno Electron microscopy in the Czech Republic http://www.isibrno.cz “Tripod,” 1950 http://www.isibrno.cz First high-vacuum system, 1961 Electron microscopy in the Czech Republic Technical Museum in BrnoTescan Factory in Brno 20 Electron microscopy of biological specimen 15um section of Drosera leaf, Marton, 1934 Bacteriophage, H. Ruska 1942 21 Electron microscopy of biological specimen Adenovirus, W. Horn T4 bacteriophage Negative staining (S. Brenner) Orf virus, W. Horn 22 Electron microscopy of biological specimen First qualitative image analysis 23 Electron microscopy of biological specimen First sub-nanometer electron microscopy structure Electron cryostallography (R. Henderson, N. Unwin) 24 Electron microscopy of biological specimen Vitrification → Cryo-electron microscopy (J. Dubochet) Aaron Klug: tim 1930 1940 1950 1960 1970 1980 1990 2000 2010 Milestones: 1968: DeRosier & Klug – first 3D EM reconstruction (phage T4) 1982: Nobel Prize in Chemistry Other notable events 1930 1940 1950 1960 1970 1980 1990 2000 2010 Milestones: 1934: Ladislaus Laszlo “Bill” Marton takes the first image of biological specimen: sections of a plant leaf 1937: Manfred von Ardenne (CRT) develops SEM 1948: Dennis Gabor develops electron holography (Nobel Prize in Physics, 1971) 1974: Ken Taylor & Bob Glaeser – electron crystallography of frozen hydrated catalase 1974: Walter Hoppe – 3D reconstruction of fatty acid synthase using tomography 1975: Richard Henderson – subnanometer electron crystallography 1982: Jacques Dubochet – modern cryo techniques 1987: Joachim Frank – “single particle” reconstruction of 50S ribosome 2007: Direct Electron develops first commercial direct electron detector Středoevropský technologický institut c/o Masarykova univerzita Žerotínovo nám. 9 601 77 Brno, Česká republika www.ceitec.cz | info@ceitec.cz