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'Instrumental Insights' ‘INSTRUMENTAL INSIGHTS’ MPI of Biochemistry: THE CAMPUS Department of Molecular Structural Biology TEM HISTORY: An Anecdote... In 1928,…. Dennis Gabor had rejected Leo Szilard’s suggestion to make a microscope based on electron waves with the response: “What is the use of it? Everything under the electron beam would burn to a cinder!” Robert M. Glaeser. Cryo-electron microscopy of biological nanostructures. Physics Today 61, 1, 48 (2008) TEM HISTORY: An Anecdote... William Lanouette. Genius in the Shadows, A Biography of Leo Szilard, the Man Behind the Bomb’; p85 TEM HISTORY: July 1938 Helmut Ruska 1908-1973 ‘Cryo-Electron Microscope’: The concept Humberto Fernández-Morán *1924 Maracaibo, Venezuela ✝1999 Stockholm, Sweden see also: Fernandez-Moran, H. (1965) Electron microscopy with high-field superconducting solenoid lenses. Proceedings of the National Academy of Sciences 53: 445-451 7 ‘Cryo-Electron Microscope’: The concept Humberto Fernández-Morán *1924 Maracaibo, Venezuela ✝1999 Stockholm, Sweden see also: Fernandez-Moran, H. (1965) Electron microscopy with high-field superconducting solenoid lenses. Proceedings of the National Academy of Sciences 53: 445-451 7 Tomography or the ‘polytropic montage’ Roger Hart Electron microscopy of unstained biological material: The polytropic montage. Science 1968, 159, 1464-1467 Tomography or the ‘polytropic montage’ Roger Hart Electron microscopy of unstained biological material: The polytropic montage. Science 1968, 159, 1464-1467 Tomography or the ‘polytropic montage’ Roger Hart Electron microscopy of unstained biological material: The polytropic montage. Science 1968, 159, 1464-1467 ‘Tomographic chicken grill’ Walter Hoppe Electron diffraction with the transmission electron microscope as a phase determining diffractometer - From spatial frequency filtering to the three-dimensional structure analysis of Ribosomes. Angewandte Chemie 1983, 22 (6), 456-485 1917-1986 Walter Hoppe Titan Krios ! Development start/a 2004 4 meters FEI Phase Plate Development end/a 2011 Voltage/kV 80-300 direct electron detectors & energy filter Samples/# 12 FEG-type Schottky Pre-GIF Lenses constant current FEI FALCON Camera Condenser 3 lenses Energy filter post-column GATAN Energyfilter Post-GIF GATAN K2 Camera …starring Mike Strauss CRYO-EM: The ‘Hurdles’ • Samples that have to be studied in their VITRIFICATION hydrated state to ensure structural converts liquid water into amorphous ice preservation • Suitable sample thickness to obtain SAMPLE PREPARATION molecular resolution ADVANCE MICROMACHINING ADVANCE 3D CORRELATION • Low-contrast due to weakly scattering POOR IMAGING CONDITIONS building blocks OPTIMIZE INSTRUMENTATION E.G. DETECTORS & PHASE PLATES • Sensitivity to ionizing radiation LOW DOSE METHODS ADVANCE AUTOMATION WHY?: Microscopy with electrons Szilard was right: At smaller and smaller electron wavelengths, one can achieve much more h h λelectrons = mv ⇡ E 2m E(1 + 2 ) 0 2m0c q energy E [keV] wavelength λ [Å] velocity v [km/sec] 20 0,086 76000 40 0,06 107000 60 0,059 131000 80 0,042 152000 100 0,037 170000 300 0,019 233000 1000 0,009 282000 ➝ Transmission electron microscopes can resolve details down to 1 Ångstrøm! TEM HISTORY: The past... 1856-1940 1897 J.J. Thomson Discovered the electron (as a ‘corpuscle’) 1924 Louis de Broglie Predicted the wavelength for the electron λ = h/mv 1926 H. Busch Magnetic and electric fields act as lenses for electrons G.P. Thomson/ 1927 C.J. Davisson and L.H. Germer Observed the wave properties by electron diffraction 1931 M. Knoll & E. Ruska Built the 1st electron microscope (EM) 1936 Metropolitan Vikers Manufactured the 1st industrial EM (EM1) 1920 Cavendish Lab, Cambridge 1938 B. von Borries & E. Ruska Produced the 1st practical EM (Siemens) – 10 nm resolution 1939 Siemens&Halske Started their commercial EM production 1962 R. Castaign & L. Henry Built the 1st energy filter 1982 P.T.E Roberts et al. Used the 1st CCD’s for EM E. Ruska/ 1986 G. Binning and H. Rohrer Received the Nobel Prize for TEM and STM J.J. Thomson (1897) "Cathode Rays", The Electrician 39, 104 and Philosophical Magazine, 44, 293. TEM HISTORY: The past... 1892-1987 1897 J.J. Thomson Discovered the electron (as a ‘corpuscle’) 1924 Louis de Broglie Predicted the wavelength for the electron λ = h/mv 1926 H. Busch Magnetic and electric fields act as lenses for electrons G.P. Thomson/ 1927 C.J. Davisson and L.H. Germer Observed the wave properties by electron diffraction 1931 M. Knoll & E. Ruska Built the 1st electron microscope (EM) 1936 Metropolitan Vikers Manufactured the 1st industrial EM (EM1) 1927 Solvay Conference, Brussels 1938 B. von Borries & E. Ruska Produced the 1st practical EM (Siemens) – 10 nm resolution 1939 Siemens&Halske Started their commercial EM production 1962 R. Castaign & L. Henry Built the 1st energy filter 1982 P.T.E Roberts et al. Used the 1st CCD’s for EM E. Ruska/ 1986 G. Binning and H. Rohrer Received the Nobel Prize for TEM and STM L. de Broglie, Recherches sur la théorie des quanta (Researches on the quantum theory), Thesis (Paris), 1924 and Ann. Phys. (Paris) 3, 22 (1925). TEM HISTORY: The past... “Father of electron optics” 1884-1973 1897 J.J. Thomson Discovered the electron (as a ‘corpuscle’) 1924 Louis de Broglie Predicted the wavelength for the electron λ = h/mv 1926 H. Busch Magnetic and electric fields act as lenses for electrons G.P. Thomson/ 1927 C.J. Davisson and L.H. Germer Observed the wave properties by electron diffraction 1931 M. Knoll & E. Ruska Built the 1st electron microscope (EM) 1936 Metropolitan Vikers Manufactured the 1st industrial EM (EM1) 1932 University of Jena 1938 B. von Borries & E. Ruska Produced the 1st practical EM (Siemens) – 10 nm resolution 1939 Siemens&Halske Started their commercial EM production 1962 R. Castaign & L. Henry Built the 1st energy filter 1982 P.T.E Roberts et al. Used the 1st CCD’s for EM E. Ruska/ 1986 G. Binning and H. Rohrer Received the Nobel Prize for TEM and STM Busch, H.: Über die Wirkungsweise der Konzentrierungsspule bei der Braunschen Röhre. (On the mode of action of the concentrating coil in the Braun tube.) Arch. Elektrotechnik 18, 583-594, Jena, Physikalisches Institut, March 1927 TEM HISTORY: The past... “Father of electron optics” 1884-1973 1897 J.J. Thomson Discovered the electron (as a ‘corpuscle’) 1924 Louis de Broglie Predicted the wavelength for the electron λ = h/mv 1926 H. Busch Magnetic and electric fields act as lenses for electrons G.P. Thomson/ 1927 C.J. Davisson and L.H. Germer Observed the wave properties by electron diffraction 1931 M. Knoll & E. Ruska Built the 1st electron microscope (EM) 1936 Metropolitan Vikers Manufactured the 1st industrial EM (EM1) 1932 University of Jena 1938 B. von Borries & E. Ruska Produced the 1st practical EM (Siemens) – 10 nm resolution 1939 Siemens&Halske Started their commercial EM production 1962 R. Castaign & L. Henry Built the 1st energy filter 1982 P.T.E Roberts et al. Used the 1st CCD’s for EM E. Ruska/ 1986 G. Binning and H. Rohrer Received the Nobel Prize for TEM and STM Busch, H.: Über die Wirkungsweise der Konzentrierungsspule bei der Braunschen Röhre. (On the mode of action of the concentrating coil in the Braun tube.) Arch. Elektrotechnik 18, 583-594, Jena, Physikalisches Institut, March 1927 TEM HISTORY: The past... 1892-1975 1897 J.J. Thomson Discovered the electron (as a ‘corpuscle’) 1924 Louis de Broglie Predicted the wavelength for the electron λ = h/mv 1926 H. Busch Magnetic and electric fields act as lenses for electrons G.P. Thomson/ 1927 C.J. Davisson and L.H. Germer Observed the wave properties by electron diffraction 1931 M. Knoll & E. Ruska Built the 1st electron microscope (EM) 1936 Metropolitan Vikers Manufactured the 1st industrial EM (EM1) 1937 Imperial College, London 1938 B. von Borries & E. Ruska Produced the 1st practical EM (Siemens) – 10 nm resolution 1939 Siemens&Halske Started their commercial EM production 1962 R. Castaign & L. Henry Built the 1st energy filter 1982 P.T.E Roberts et al. Used the 1st CCD’s for EM 1937 Bell Telephone Labs, USA E. Ruska/ 1986 G. Binning and H. Rohrer Received the Nobel Prize for TEM and STM C. J. Davisson and L. H. Germer, Nature 119, 558 (1927). and G. P. Thomson and A. Reid, Nature 119, 890 (1927). TEM HISTORY: The past... 1897 J.J. Thomson Discovered the electron (as a ‘corpuscle’) 1924 Louis de Broglie Predicted the wavelength for the electron λ = h/mv 1926 H. Busch Magnetic and electric fields act as lenses for electrons G.P. Thomson/ 1927 C.J. Davisson and L.H. Germer Observed the wave properties by electron diffraction 1931 M. Knoll & E. Ruska Built the 1st electron microscope (EM) 1936 Metropolitan Vikers Manufactured the 1st industrial EM (EM1) 1931 Technical University, Berlin 1938 B. von Borries & E. Ruska Produced the 1st practical EM (Siemens) – 10 nm resolution 1939 Siemens&Halske Started their commercial EM production 1962 R. Castaign & L. Henry Built the 1st energy filter 1982 P.T.E Roberts et al. Used the 1st CCD’s for EM E. Ruska/ 1986 G. Binning and H. Rohrer Received the Nobel Prize for TEM and STM E. Ruska und M. Knoll: Die magnetische Sammelspule für schnelle Elektronenstrahlen. Z. techn. Physik. Band 12, 1931, 389–400 und 448 and M. Knoll und E. Ruska: Das Elektronenmikroskop. Z. Physik 78, 1932, 318-339. TEM HISTORY: The past... 1906-1988 1897 J.J. Thomson Discovered the electron (as a ‘corpuscle’) 1924 Louis de Broglie Predicted the wavelength for the electron λ = h/mv 1926 H. Busch Magnetic and electric fields act as lenses for electrons G.P. Thomson/ 1927 C.J. Davisson and L.H. Germer Observed the wave properties by electron diffraction 1931 M. Knoll & E. Ruska Built the 1st electron microscope (EM) 1936 Metropolitan Vikers Manufactured the 1st industrial EM (EM1) 1939 Technical University, Berlin 1938 B.
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