The main theme underlying this approach is the excitation of fluorescent molecules followed by the action of a high power Depletion Laser. The depletion laser is instrumental in depleting the excited state of the molecules in the periphery of the focal spot by stimulated emission, thereby reducing the point spread function (PSF), leading to higher resolution.

STED and STELLARIS as a single system offers a briliant confocal imaging with unique super resolution capabilities to help you dive progress in science. Acquire outstanding confocal and STED image easily and set up your experiment in a few click. Thanks to the combination with the STELLARIS platform, STED benefits from the Leica next generation White Light Laser (WLL) / AOBS technology, a newly optimized beam path, and spectral detection driven by the Power hyd detector family. Together with the availability of 775 STED line, STED enables you to characterize structures with nanoscale detail, perform colocalization studies with the highest flexibility in terms of fluorophore selection, and follow highly dynamic processes.

Expand the potential of STED experiments with outstanding resolution, image quality, and sample protection provided by our proprietary tau-STED technology. Tau-STED combines the optical signals from STED and the physical information from the fluorescence lifetime at confocal speeds. This approach uses phasor analysis in a novel way that delivers outstanding STED resolution* and image quality, while removing background noise, even at low excitation and STED power. You can apply tau-STED for all your STED experiments, in particular for multicolor colocalization studies and when you need to follow fast processes in live specimens, for extended periods, or over large volumes.

Ability to validate your results fast and on a single platform: The combination of STED and STELLARIS means you can validate your findings using different dimensions of data from your specimen. STED, confocal, LIGHTNING, and tausense data are all available with the same system. You can access your raw data for quantification.

Sample Carriers

The sample carriers may include live cell imaging dishes, Multichamber , and fix slides.

Applications

1. STED: Super-resolution Imaging:

STED functions by depleting fluorescence in specific regions of the sample while leaving a center focal spot active to emit fluorescence. This focal area can be engineered by altering the properties of the pupil plane of the objective lens. In principle, the higher the power of the depletion laser, the more is the reduction in the point spread function.

2. Multicolor/Colocalization imaging:

Multicolor STED was developed in response to a growing problem in using STED to study the dependency between structure and function in proteins. To study this type of complex system, at least two separate fluorophores must be used. Using two fluorescent dyes and beam pairs, colocalized imaging of synaptic and mitochondrial protein clusters is possible.

3. Tau-STED imaging:

Tau-STED is a super-resolution microscopy technique that combines fluorescence lifetime measurements with STED (stimulated emission depletion) imaging. It allows for the imaging of live cells and intact specimens at a nanoscale resolution. Tau-STED can be used to image live cells and intact specimens. It can be used to resolve small details in context at low light dose. It can provide multicolor imaging having single color excitation and variant life time cluster.

4. FRET/Tau-FRET:

Fluorescence resonance energy transfer (FRET) is a technique that measures the transfer of energy between two fluorophores. It's a spectroscopic technique that's used to study molecular interactions, such as protein-protein interactions and biomolecular dynamics. Using Tau FRET, image can be acquired based on contrast life time dependency in fluorescence region of FRET pair interaction.

5. FRAP:

Fluorescence recovery after photobleaching (FRAP) is a cutting-edge live-cell functional imaging technique that enable the exploration of biomolecular dynamics such as protein in individual cells and thus permits the elucidation of protein mobility, function and interaction at a single –cell level. A technique used to study diffusion and interactions of macromolecules, hence it can be used to determine whether a sensor is freely diffusing or if it is confined within specific regions of the cell.

6. Live cell imaging:

Live-cell labeling with fluorescent proteins: Useful for thicker sections and live-cell imaging, targeted intracellular labels.