STED Microscopy

Understanding protein nano distribution requires the use of super resolution technique.

STED (STimulated Emission Depletion) microscopy technique belongs to the super resolution microscopy. Our STED microscope allows to image beyond the diffraction limits in 2D and 3D.

The principle of STED microscopy consists in the superposition between an excitation laser and a depletion laser having the shape of a donut. The depletion leads to the passage of the flurochromes in OFF state. The temporal and spatial synchronization of these two lasers leads to the reduction of the point spread function (PSF) of our optical system and thus to the improvement of the resolution.

Thanks to the combination of the STED with the FALCON module allowing to measure lifetime, we can sort the photons according to their lifetime to have a more accurate selection of “useful” photons.

It also allows to decrease the background noise of the sample, so the signal to noise ratio is considerably improved. The Tau-STED can allow us to decrease the power of depletion laser and thus be more compatible with the live imaging.

Contact: Magali Mondin / Christel Poujol

Leica Microsystems

3D reconstruction of a dendrite inside a brain slice of Thy-1 GFP mouse. The GFP has been detected by a immuno-labelling using Alexa 647

Julie Angibaud, IINS UMR 5797 CNRS

Cis-golgi (Green) and trans-golgi (Magenta) markers in root from arabidopsis thaliana

Magali Grison, UMR 5200 CNRS Université Bordeaux