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.
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:
Taekjip Ha, Illinois Department of Physics
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.
Paul Selvin, Illinois Department of Physics
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).