Syllabus database for doctoral courses
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Syllabus database for doctoral courses
SYLLABI FOR DOCTORAL COURSES
| Swedish title | Funktionell Fluorescens Mikroscopi Avbildning (fFMA) i biomedicinsk forskning |
|---|---|
| English title | Functional Fluorescence Microscopy Imaging (fFMI) in biomedical research |
| Course number | 2348 |
| Credits | 3.0 |
| Responsible KI department | Institutionen för klinisk neurovetenskap |
| Specific entry requirements | |
| Grading | Passed /Not passed |
| Established by | The Board of Doctoral Education |
| Established | 2009-09-15 |
| Purpose of the course | |
| Intended learning outcomes | At the end of the course the student should be able: 1. to use fundamental aspects of molecular structure to describe molecule (matter) interaction with light (electromagnetic radiation) and the emission of fluorescence, and to use this knowledge to discuss fluorescent properties of a fluorophore and different labeling strategies. 2. to understand the fluorescence imaging instrumentation, to define elements of a fluorescent microscope and to describe instrumentation properties in terms of sensitivity. 3. to describe the theoretical background behind fluorescence based methodologies for studying molecular interactions in live cells. Discuss pros and contras in relation to the biological problem studied. 4. to specify instrumental requirements and to design a fluorescence imaging assay for a biological problem of interest. 5. to apply a specific labeling strategy and perform a live cell fluorescence imaging assay. 6. to document experimental work in accordance with Good Laboratory Practice (GLP) guidelines. 7. to communicate the results in written and oral form. 8. to discuss the adequateness of the methodology used in the scientific literature concerned. |
| Contents of the course | Fluorescence microscopy and associated techniques are becoming indispensable research tools for investigating molecular mechanisms of biological processes. Versatility of fluorescence based methodologies comes from the multifaceted possibility to characterize fluorescence emission by position, intensity, wavelength, lifetime and polarization. These methodologies bring new biological insight at an unprecedented rate and are of crucial importance for the development of life sciences. The course covers the following topics: Luminescence and the nature of light (Fluorescence, Phosphorescence, Light scattering, Rayleigh-Tyndall scattering, Raman scattering); Fluorescent markers and their photophysical properties (Organic fluorescent dyes for covalent conjugation (Rhodamine 6G, Alexa dyes, Cyanine dyes); Quantum dots; Intrinsically Fluorescent Proteins (Aequorea victoria (GFP, YFP), Discosoma coral (DsRFP) and Montipora (Keima) families); Selectively binding and intercalating dyes (DiI, DraQ 5). Labeling strategies for biomedical application. Instrumentation for fluorescence imaging (Light sources, Wavelength selection, Detectors, Read-out devices, Sample holders, Problems of high blank values, Cuvettes, Solvents and Reagents, Other contaminants); Quantitation and Sensitivity (Instrumental sensitivity, Method sensitivity, Absolute sensitivity); Factors affecting quantitative accuracy (Adsorption, Photo-decomposition, Oxidation, Temperature, pH, Inner-filter effects, Quenching); Fluorescence based methodologies for studying molecular interactions in live cells (FRET, FLIM, FRAP). Single-molecule detection (Quantitative APD imaging, FCS and FCCS). Non-linear microscopy (Multi-photon excitation, STED). Spatially resolved fluorescence imaging (STORM, PALM). |
| Teaching and learning activities | The course includes lectures, laboratory sessions, discussion sessions and a project work. |
| Compulsory elements | All sessions are compulsory. Please report any absence to the course leader in advance by e-mail. Absence from any part of the course (lectures, laboratory sessions, discussion sessions and exam) is generally not accepted but could in special cases be compensated by an individually tailored additional module and a special written examination organized by the course committee. |
| Examination | The final assignment consists of a written project report (5 pages) and an oral presentation of the project report (15 min). |
| Literature and other teaching material | Selected chapters from: Joseph R. Lakowicz, Principles of Fluorescence Spectroscopy, Springer, 2006. Mary-Ann Mycek & Brian W. Pogue: Handbook of Biomedical Fluorescence, CRC, 2003. On-line virtual microscopy interactive Java tutorial (http://www.olympusconfocal.com/java/index.html) Fluorescence Correlation Spectroscopy (FCS) and Fluorescence Cross-Correlation Spectroscopy (FCCS) http://fcsxpert.com/classroom/ Handouts: Scientific papers with fFMI methodology |
Course responsible |
Vladana Vukojevic Institutionen för klinisk neurovetenskap 51771797 0703060648 Vladana.Vukojevic@ki.se CMM L8:01 17176 Stockholm |
| Contact person |
