| Responsible Department | Department of Plant Biology and Biotechnology | ||||||||||||||
| Course Dates | as agreed upon | ||||||||||||||
| Course Abstract | The course covers all biologically relevant methods in microscopy and presents their theoretical background as well as the practical control of the instruments. Preparations methods and training at the instruments is included. The course is structured in modules related to the different instruments and methods. Students interested in the course can freely choose the appropriate module(s). | ||||||||||||||
| Course Registration | To sign up for the course, please send an e-mail to Alexander Schulz at als@life.ku.dk. Please also remember to add the course to your PhD plan | ||||||||||||||
| Deadline for Registration | as agreed upon | ||||||||||||||
| Credits | 6 (ECTS) | ||||||||||||||
| Level of Course | PhD course | ||||||||||||||
| Language of Instruction | English may be conducted in Danish | ||||||||||||||
| Restrictions | Module a: 8 students, Modules b-f 2-6 | ||||||||||||||
| Course Content | |||||||||||||||
| General: For all the modules described below, the course is based on (i) Literature studies, (ii) Laboratory work, resp. work at the microscope, (iii) Supervision of practical and theoretical work, and (iv) Final submission of a written report on the laboratory work including a discussion which refers to the latest relevant literature on the selected subject. Module a: Light Microscopy (6 ECTS) This module covers all light microscopic methods (bright field, phase contrast, fluorescence, polarisation, differential contrast, confocal). The optical and technical background is thoroughly discussed (resolution, Köhler illumination, wave optics, diffraction, amplitude and phase specimen birefringence), and measuring methods and the effects of different instrument settings are practiced. The students are requested to come with their own specimen and will learn how these can optimally be documented. This course module will run when at least three students participate. Because of the limited number of microscopes suited, the number of participants is limited to six. Module b: Histology (6 ECTS) This module is designed according to the aim of the student's wishes or actual research project and includes theoretical as well as practical training in histological sample preparation of plant material and interpretation of the resulting image. Suggested subjects: (1) whole mount sampling, (2) procedures for plant tissue sections for light microscopy (incl. fixation, embedding, sectioning and staining) (3) Staining and histochemistry, including the theoretical background and identification of potential artifacts. Module c: Confocal Laser Scanning Microscopy CLSM (3-6 ECTS) The course module is designed either as a general introduction to confocal microscopy (3 ECTS) or will include a project related to the Ph. D. student's actual research project (6 ECTS). In both cases the module will include theoretical as well as practical training. One or more of following topics are covered: principles of confocal and two-photon microscopy; staining and mounting of living or fixed specimens; operating the microscope for 3D, time lapse, or flow chamber recordings; image analysis of the obtained data. Module d: Scanning Electron Microscopy SEM (3-6 ETCS) The course module is designed either as a general introduction to scanning electron microscopy (3 ECTS) or will include a project related to the Ph. D. student's actual research project (6 ECTS). In both cases the module will include theoretical as well as practical training. Following topics are covered: principles of SEM and ESEM (environmental scanning electron microscopy); fixation, dehydration, and coating of specimen; operation the instrument for SEM and ESEM. Module e: Transmission Electron Microscopy TEM (6 ECTS) The course module will include a project related to the Ph. D student's actual research project. The theoretical and practical implications of TEM preparation (fixation, sectioning, contrasting) will be treated. The student will learn to make ultrathin sections, to contrast them and how to operate the electron microscope. For documentation, both analogue (plane film) and digital imaging will be introduced. Module f: Fluorescence Microscope FM (3 ECTS) Starting point for this course module is a specimen from the student's Ph. D. project, e.g., a GFP-transformed plant. Screening for GFP-positive lines at the stereo fluorescence microscope, preparation of specimen for the fluorescence microscope, selection of filter combinations and documentation of fluorescence micrographs with the digital camera are parts of the course module. | |||||||||||||||
| Teaching and learning Methods | |||||||||||||||
| Lectures on and discussions of the theoretical background of the relevant methods. Practical training in preparation and training at the instruments used in the single modules. In module a Journal Club of relevant articles. | |||||||||||||||
| Learning Outcome | |||||||||||||||
| Knowledge Understanding of the theoretical background of the microscopes used in the respective module, such as preparation and fixation protocols matching different organisms, optical geometry and image formation of the microscope(s), influence of radiation on specimens and artefacts, and image capturing. Skills Control the basic and advanced settings of the microscopes and cameras used, learn to control the stage and to focus the instrument to the region of interest, evaluate and improve the image quality (filter settings, brightness, contrast). Mount images to compound panels, determine the magnification and label figures with magnification bar, letters, and arrows. Competences Professional use of the microscopes used in the different modules. Capability to discriminate artefacts from real structures. Determine the best instrument to document the structures in question. | |||||||||||||||
| Course Material | |||||||||||||||
| For module a: Manual: 'Light Microscopic Methods - PhD course' by Alexander Schulz | |||||||||||||||
| Course Coordinator | |||||||||||||||
| Alexander Schulz, als@life.ku.dk, Department of Plant Biology and Biotechnology/Section for Transport Biology, Phone: 353-33350 Michael Hansen, mh@life.ku.dk, Department of Plant Biology and Biotechnology/Section for Transport Biology, Phone: 353-33331 Helle Juel Martens, hjm@life.ku.dk, Department of Plant Biology and Biotechnology/Section for Plant Biochemistry, Phone: 353-32193 | |||||||||||||||
| Course Fee | |||||||||||||||
| There is no course fee for PhD students from Danish universities. All others must pay a fee of DKK 4000 | |||||||||||||||
| Course Costs | |||||||||||||||
| Bench fees: The students have to expect moderate bench fees for chemicals used in module b, c, d, and e | |||||||||||||||
| Type of Evaluation | |||||||||||||||
| Final submission of a written report on the laboratory work including a discussion which refers to the latest relevant literature on the selected subject. | |||||||||||||||
| Work Load | |||||||||||||||
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