2 November 2007
Saveez Saffarian
Kirchhausen Lab
HMS
I will summarize a burst of high resolution methods which are based on fluorescence imaging of fixed samples. Most of these techniques rely on measurements of the center of fluorescence of individual fluorescent molecules. I will show how we used these concepts in live cells with a new method named DiNa (Differential Evanescence nanometry) with which we have achieved a 10nm axial resolution near the plasma membrane.
Clathrin coated pits and coated vesicles form at the plasma membrane of eukaryotic cells. The assembly dynamics of these diffraction-limited objects has been followed in cells expressing fluorescent chimeras of clathrin and its AP-2 adaptor complex. I will present our measurements of the position of the adaptors within the forming clathrin coated vesicle using DiNa. The vesicles have a final diameter of ~100nm.