Electron microscopic techniques of plants I
1. Basics. History of light and electron microscopy, fundamentals of optics, light microscopy, electron beam, adventages of electron microscopy.
2. Principals for conventional TEM I. Sampling, size of sample, fixatives, vehicles for fixatives, dehydration, infiltration, embedding, resins and their features.
3. Principals for conventional TEM II. Artifacts during sampling, fixation, dehydration and embedding. Semithin and thin sections, grids, coating materials and their characteristics.
4. Principals for conventional TEM III. Knifes, ultramicrotomes, artifacts in sectioning. Staining, negative staining, replica techiques, castings, autoradiography
5. Preparation of biological samples for SEM Electron beam - specimen interactions, information from SEM, sampling, fixation, dehydration, drying of samples (critical point drying and freeze drying), coating (vacuum and sputter coating), artifacts in SEM sampling
6. Cryoelectron microscopy Cryofixation, freeze-fracture, freeze-etching, freeze-drying, freeze-substitution, LT TEM (cryosection, cryotransfer), LT SEM
7. Electron microscopic immunolabelling techniques Antigenes, antibodies, specificity, cross reaction, markers, direct and indirect labelling, protein enginering, antibody planning and synthesis
8. Diffraction in electron microscopy Diffraction, information from diffraction, diffraction on biological samples
9. Transmission electron microscope Principle of TEM, basics of structure and work, resolution, contrast, artifacts of TEM operation and their consequenses, special TEM techniques.
10. Scanning electron microscopy Principle of SEM, basics of structure and work, resolution, contrast, influence of settings on image quality, artifacts of SEM operation and their consequenses, special SEM techniques.
11. Documentation Analogous photography, films, developers, paphotographic papers, sensitivity, speed, contras, tonal gradation, handling, digital photography, image processing, artifacts in photographical work.
12. X-ray microanalysis Principles, signals from inelasting scattering, X-rays, detectors, basic design, sensitivity, WDS, EDS, sample prepare specialities, analysis, artifacts of the detection process, spectral artifacts
13. Electron energy loss spectroscopy (EELS) Principles, electron energy loss, detectation, sensitivity, basic design, sample prepare specialities, analysis, spectra, artifacts
14. Electron microscopic tomography, image processing 3D, tomography, basics of image processing, methods for image quality improvement, image modification (contrast, labels, restoration, fals coloring), measurements, morphometrical applications
15. Special applications of TEM and SEM Exeptional technical applications, unusual objects and experiments from literature.
J. L. Hall and C. Hawes: Electron microscopy of plant Cell, Academic Press, London, 1991.