Lect 2- Techniques II

Lect 2- Techniques II - Lecture 2: Cell biological...

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Unformatted text preview: Lecture 2: Cell biological Techniques II Make sure that you have access to Blackboard. If you do not have access to blackboard, you are probably not registered for the course. A new version of the syllabus with corrections to discussion sections and final exam day has been posted. FRET Fluorescence Resonance Energy Transfer (FRET): Used to show not just co-localization (200 nm optical limit), but direct interactions. If two fluorophors are within ~100 angstroms (10 nm, ~ the size of proteins and macromolecular complexes), they can transfer energy to each other. Energy transfer is dependent on [distance]1/6, so distances must be very small. Illuminate at absorbance peak of one fluorophor, and measure fluorescence at emission of the second. The emission of the first fluorophor must overlap with the absorbance of the second fluorophor to transfer energy. FRET Illuminate at absorbance peak of one fluorophor, and measure fluorescence at emission of the second. The emission of the first fluorophor must overlap with the absorbance of the second fluorophor to transfer energy. FRET Illuminate at absorbance peak of one fluorophor, and measure fluorescence at emission of the second. fluorescence at emission of the second....
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Lect 2- Techniques II - Lecture 2: Cell biological...

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