CLB-30306 Cell Biology and Advanced Imaging Technologies

Course

Credits 6.00

Language of instruction:

English

Assumed knowledge on:

CBI-10306 Cell Biology, MAT-15403 Statistics 2, PCC-12803 General Chemistry for the Life Sciences, MAT-14903 Mathematics 2 or equivalent courses. Basic knowledge of optics (lens equation) and electromagnetic waves (wavelength, phase, photons and absorption).

Continuation courses:

Thesis CLB, BIC, PPH or MIB; MOB-30806 Regulation of Plant Development.

Contents:

Note: This course has a maximum number of participants. The deadline for registration is one week earlier than usual. See Academic Year (http://www.wur.nl/en/Education-Programmes/Current-Students/Agenda-Calendar-Academic-Year.htm) -> Registration for Courses.
Advanced light microscopy and reporter gene techniques are indispensable for modern days mechanistic studies on cellular functioning and organisation. New microscopy technologies and digital imaging allow for studies of dynamic processes at shorter time scales and with higher spatial resolution in living cells. In this practical course the principles and cell biological application of different microscopy techniques will be taught. Topics include (green) fluorescent proteins (GFP), luciferase reporter activity, widefield and confocal fluorescence microscopy, differential interference contrast microscopy (DIC), fluorescence lifetime imaging microscopy (FLIM), Förster resonance energy transfer (FRET), fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP), digital imaging, image noise, and microscope construction. These concepts will be applied during the course to study promoter activity during hormone treatment; dynamics of intracellular organisation; protein diffusion; circadian rhythm in plants; and intermolecular interactions. An important aspect of the course is data acquisition, reduction and analysis. How can quantitative data be extracted from digital images for comparison with quantitative models on molecular interactions and cellular organization?
Note:
This is a course with a maximal capacity of 40 students. Students are accepted in the order of: 1) students for which this course is compulsory; 2) students for which this course is restricted optional in their programme; 3) students for which the course is part of their minor; 4) students for which this is a free choice course. The last open spots are given to students in order of their registration. The deadline for course registration is one week before the deadline of other courses.

Learning outcomes:

After successful completion of this course students are expected to be able to:
- explain principles of light microscopy techniques and reporter methods;
- identify the components of advanced light microscopes and know the basics of their operation;
- operate light microscopes based on a knowledge of their design;
- extract quantitative data from digital images;
- process data and compare results with provided quantitative models on molecular and cell biological processes;
- critically evaluate the use of light microscopy and analysis methods in literature;
- write a short research paper on a conducted cell biological experiment and present the work orally;
- discuss the presented cell biological processes in a mechanistic manner;
- select appropriate microscopy techniques for cell biological work.

Activities:

Lectures. Conduct experiments on cells and biochemical reconstitution assays using advanced light microscopes (experiments are compulsory since acquired skills are part of the learning outcome). Use a computer module to actively simulate the use of fluorescence microscopes. Use computer programs (Excel, imageJ/Fiji) for data handling and processing. Study cell biological research papers that apply light microscopy and participation in literature discussion groups. Write a report in the form of a research paper and present the work with an oral presentation.

Examination:

- written exam with mainly open questions that assess your understanding of the lecture topics and concepts of the practical experiments (70%). The written exam needs a minimal mark of 5.5 to pass;
- a report on one of the performed practical experiments, written in teams of 2 students, is assessed for style of writing, line of argumentation, clarity of presentation, incorporation of study material from the course, and discussion (20%);
- a mark is given for the oral presentation on the findings of the report (5%;)
- participation and achievements during the experiments and literature discussions are judged (5%).

Literature:

A syllabus will be provided at the first day of the course. Eduweb pages contain lecture slides, background literature, links to online material, and student reports on the conducted experiments.