The aim of the talk is to introduce MicroTime 200 as the all-in-one platform for time resolved microscopy with single molecule sensitivity. Modern time-resolved spectroscopic and microscopic measurements with single-molecule sensitivity allows to monitor fluorescence dynamics from sub-nanosecond to fluorescence fluctuations over seconds. Here we will explain the underlying data acquisition technique (Time-Tagged Time-Resolved (TTTR) recording) which allows to store individually timing as well as spectral and polarization information for every photon, based on a multichannel detection scheme. Having all available information with picosecond time resolution at hand, the recorded data can be analyzed in a manifold manner, employing complementary analysis schemes like fluorescence lifetime imaging (FLIM), Fluorescence Correlation Spectroscopy (FCS) and anisotropy imaging. Emphasis will be given on how time resolved measurements could be employed for monitoring Förster Resonance Transfer (FRET) related assays.

As the latest add-on to the MicroTime 200 platform, STED super-resolution microscopy extends the range of possible measurement modalities such as single molecule spectrography and atomic force microscopy (AFM). Since it is based on the time-resolved confocal platform, STED benefits from the data acquistion mode TTTR as well as high-end electronics for laser control and data collection. The easySTED approach which we employ makes the system very robust and easy to use.


Seminar, May 22, 2015, 11:00. Seminar Room

Hosted by Prof. Maria García-Parajo