The W.M. Keck Center for Cellular Imaging, a University Imaging Center is sponsoring an advanced course on fluorescence resonance energy transfer (FRET) microscopy. The workshop will be an intensive lecture/laboratory series that will enable participants to develop theoretical and hands-on expertise with various FRET microscopy techniques. Attendees will closely interact with the teaching faculty and assistants while using modern research grade microscopes equipped with essential filters, objective lenses, high-sensitivity detectors (CCD cameras and PMTs), and state-of-the-art user friendly software. This four days intensive course includes morning lectures and afternoon hands on training on FRET microscopy systems. Experienced staff from universities and industry will provide instruction. With a maximum enrollment of 20 students, there will be ample opportunity to the attendees can work with various state-of-the-art FRET microscopes, video cameras, recorders, and image processing software.
Topics to be covered include:
Fluorescence microscopy, confocal laser scanning microscopy, multiphoton excitation fluorescence microscopy and digital deconvolution and 3-D reconstruction, fluorescence lifetime imaging microscopy
Basics of fluorescence, theory of resonance energy transfer (RET), existing techniques for RET detection, need of microscopy for FRET imaging
Principles of microscope design and image formation, detectors for video imaging
Fluorophore pairs for FRET, Green fluorescent proteins, conventional fluorophores, fluorophore labeling conditions for various biological applications.
Selection of filters for FRET imaging, various FRET microscopy techniques, positive and negative controls, photobleaching FRET, FRET data analysis
Localization of proteins in live and fixed cells
The workshop is designed primarily for research scientists, physicians, postdoctoral trainees and graduate students in medical, animal, and plant research. Non-biologists seeking a comprehensive introduction to FRET microscopy will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of microscopy and molecular biology is desirable.
The five-day course consists of lectures, training on various FRET microscopy, exercises, discussions, and presentation by the attendees that will enable the attendees to obtain and interpret FRET images of high quality, to understand the importance of qualitative and quantitative FRET data analysis.