The course "Transmission Electron Microscopy in Life Sciences" is aimed at beginners and intermediate users of transmission electron microscopes in biomedicine. It will devote about equal time to the theory and to practical use of microscopes. The course is limited to 15 participants and during practical sessions the participants will be divided into three groups. The techniques discussed at theoretical sessions will be demonstrated on three transmission electron microscopes of varying complexity - the simplest JEM-1400FLASH, more complex Philips CM 100 and the mostpowerful JEM-F200 "F2". After the course, the participants should understand the principles of construction and function of transmission electron microscopes, should be able to align the microscope for optimal performance, to identify and eliminate most common aberrations and alignment artifacts, to understand the principles of image generation in transmission electron microscopy, and to identify optimal ways of image acquisition. The participants will be also given up-to-date information about best ways of sample preparation for transmission electron microscopy, and about recent transmission electron microscopy trends in biomedicine. Participants are encouraged to bring their own samples on 3.05 mm TEM grids. There will be ample opportunity for the participants to discuss their specific problems with the faculty. The course will be taught in English.
The course is organized in the framework of the project No.
TE01020118 "Electron microscopy" supported by TACR.
Hozák (Institute of Molecular Genetics of the CAS, Prague)
Vlada Filimonenko Ph.D. (Institute of Molecular Genetics of the CAS, Prague)
Microscopy Centre - Core Facility for Electron Microscopy
Institute of Molecular
Genetics of the CAS, Vídeňská 1083, Prague 4 - Krč, Czech Republic
I. Theoretical background
Introduction to transmission electron microscopy, general construction of transmission electron microscopes; filament (Wehnelt), apertures, lenses, Cs, Cc and focal length, high tension; alignments, TEM imaging bright/dark field, resolution, in-focus, over and under focus, astigmatism (condenser, objective and diffraction astigmatism), contrast; digital cameras; filter technology; the highest resolution in Life Sciences, best practice in biology; sample preparation and cryo-electron microscopy. Future prospects of TEM in biomedicine.
II. Practical sessions
Apertures, condenser and objective apertures, SA-aperture;
adjusting the filament, demonstration of bias, C1 and C2 lenses, lifetime of filament, consequences of high bias and current; specimen eucentric position, focusing the microscope, wobbler usage; optimizing brightness (light), high tension; correcting astigmatism, the methods used on minimum contrast used by material scientists and the Hole method in combination with under-focus/over-focus;
direct alignments, alignment procedures, test specimens;
dark field (conical), wobbler adjustment.
The course is organized by the Microscopy Centre – Core Facility for Electron Microscopy, which is supported from the program for large research infrastructures of the Ministry of Education, Youth and Sports within the project “National Infrastructure for Biological and Medical Imaging (Czech-BioImaging – LM2015062)“.