Dr. Timo Zimmermann
Head of the Advanced Light Microscopy Unit,
CRG – Centre for Genomic Regulation,
Biological applications for the next generation of STED microscopy systems
Stimulated Emission Depletion (STED) constitutes a powerful method for resolution improvement beyond the diffraction in light microscopy.
Additional time-gating (gSTED) of the detection signal can be used in systems with continuous wave (CW) laser depletion to improve the image resolution even further.
Until now, these improvements were limited in gated CW-STED systems to the lateral resolution of the image and to the depletion of green fluorophores at 592 nm. The now introduced possibility to improve axial resolution and the introduction of an additional laser line for depletion at 660 nm offer new ways to image biological samples. We have taken highly resolved 3D and multichannel datasets of a variety of specimens and structures to make full use of these new features.
In the talk, several projects will be covered that make use of the improved z-resolution and that use orange to red dyes and dye combinations for the new depletion laser line.
Dr. Eric Hosy
University of Bordeaux,
Live cell localization microscopy with Leica SR GSD 3D
Brownian diffusion tends to homogenize proteins that should diffuse rapidly inside the cytoplasm or cell surface. Protein clustering or immobilization necessitates specific molecular organization which counteracts this phenomenon.
During the last ten years, knowledge about the role and properties of protein diffusion made a big step forward thanks to the development of single molecule and Quantum dot tracking (SPT). However the sparse labelling inherent to these techniques render it impossible to give an overview of the protein organization.
We will report, in this webinar, the historic, the interest and the proper use of a new optical technique allowing users to obtain a high number of single molecule trajectories at the surface of a living cell using the Leica SR GSD 3D system. This versatile method, developed in 2010 and called U-PAINT, is suitable for studying any membrane biomolecule (endogenous or transfected) and provides dynamical information with large statistics revealing localization specific diffusion properties of a molecule on a single cell.