Index
Training Description
Center for the Physics of Living Cells
2012 UIUC Physical Biology Boot Camp
on Single-Molecule Fluorescence
January 11-13, 2012
Basic Training
Introductory training will include lectures from Physics Professors, Taekjip Ha and Paul Selvin, hands-on training in Ha and Selvin labs on how to build a TIRF excitation set-up.
Experimental Module
All students will learn to carry out both smFRET and FIONA experiments in the Ha and Selvin labs. This will include slide set-up, carrying out experiments, data collection and analysis, and student presentations. Descriptions of the two experimental systems you will learn are provided below:
Single-Molecule FRET
Taekjip Ha, Illinois Department of Physics
http://bio.physics.illinois.edu/
Fluorescence Resonance Energy Transfer (FRET) at the single molecule level is a powerful method to reveal conformational changes and dynamics of single biological molecules and intermolecular reactions during biochemical reactions in real time. Students will learn total internal reflection fluorescence microscopy based single molecule FRET methods using several interesting biological molecules such as the Holliday junction, DNA helicase, and RNA Polymerase.
Single-Molecule FIONA
Paul Selvin, Illinois Department of Physics
http://people.physics.illinois.edu/Selvin/PRS/PRS.html
We will explore the basics of Fluorescence Imaging with One Nanometer Accuracy (FIONA). We will watch a molecular motor (kinesin, myosin) walk on its highway (microtubules, actin, respectively). We will also extend this to in vivo FIONA, looking at the stoichiometry of various receptors (such as the nAChR or AMPAR).