STED (STimulated Emission Depletion) microscopy provides fast, intuitive, and purely optical access to study subcellular architecture and dynamics at the nanoscale. STED super-resolution meets the requirements of daily research and enables to discover minute details with live cell imaging capabilities. TCS SP8 STED 3X, allows the whole spectrum of visible light and opens the door to super-resolution in all dimensions.
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Super-resolution in all dimensions – With TCS SP8 STED 3X you can choose best resolution in xy and z and engineer the PSF to your needs.
Super-resolution over the whole spectrum – TCS SP8 STED 3X offers multiple STED laser lines at the instrument. A broad range of fluorophores become accessible.
With TCS SP8 STED 3X, two light paths are available at the instrument that generate different STED patterns. The vortex phase mask creates the classic STED donut which improves resolution only in x and y.
A second light path with a different phase mask forms the z donut that increases resolution also in z. STED light can be steplessly distributed between the two paths.
You can choose between best lateral resolution, best vertical resolution or anything in between to get optimal results. Ultra-thin optical sections reveal unknown details. TCS SP8 STED 3X gives you the possibility to match the resolution of your microscope in all dimensions to your question and specimen.
Resolution improvements by allocating the light to the two STED pathways: best lateral resolution is achieved by the classic vortex donut, best vertical resolution by the novel z donut. It can also be optimized for smallest focal volume.
Multicolor applications give access to detailed information about the interrelationships of various structures. The STED 3X module offers multiple STED laser lines in one instrument: two continuous wave lasers at 592 nm and 660 nm and a pulsed laser at 775 nm cover the whole spectrum of visible light and give access to many applicable fluorophores.
The White Light Laser, the AOBS (acousto optical beam splitter) and the tunable spectral detector synergistically enable you to image any kind of fluorophore combination and give you the highest flexibility for your multicolor super-resolution experiment.
Gated STED substantially extends the functionality of the proven STED CW, giving you the option of higher resolution or lower laser power. More images are obtained and smaller details revealed.
In STED CW imaging resolution is highly dependent on the lifetime of the fluorophore. The STED laser silences fluorophores with long lifetimes more efficiently than short-living ones. The longer a fluorophore stays on average in the excited state, the higher is the chance for stimulated emission and therefore the better the resolution. The lifetime is not evenly distributed at the focal spot and long-living states are found in the center, whereas short-living states are located at the periphery where the STED laser is active. Observing only the long-lived states shrinks the effective spot further.
The HyD™ detectors together with the White Light Laser as pulsed excitation source give the possibility to detect only in a certain time gate after the excitation pulse. By shifting the time gate away from the excitation pulse a resolution far below 50 nm can be reached. With the same laser power, gated STED achieves a more than 50% higher resolution than STED CW. Smaller details can be observed without the need to apply more STED light, increasing live cell capabilities.
STED microscopy is the fast and direct way to super-resolution. The LAS AF (Leica Application Suite Advanced Fluorescence) grants you the greatest possible comfort. A sketch of the estimated effective PSF gives you direct and online visual feedback on the effects of your chosen technical parameters.
As an additional innovation the Smart STED Wizard is implemented into LAS AF. The intuitive workflow allows you to control the instrument with three simple sliders. Assisted by the sketch of the effective PSF you define the general level of super-resolution you need. For optimal outcome the wizard adjusts all necessary settings like STED laser intensity, pixel size, z step size, the pinhole, gate settings and averaging.
With every TCS SP8 STED 3X system you exclusively get a powerful Huygens STED deconvolution package by SVI. Huygens increases contrast dramatically and also enhances resolution in x, y and z. The outcome can be directly compared with the raw data and avoids being mislead by image processing artifacts. Leica and SVI have enabled Huygens to handle 3D STED data and have developed LAS AF<->Huygens data exchange that facilitates the interplay of the two software packages. One mouse click sends acquired data to Huygens for deconvolution. Vice versa, deconvoluted pictures are just as easily sent back to LAS AF.
Objectives are the eyes of every microscope and critical for determining the resolving power of a confocal system. Transmission and color correction of an objective influence excitation and detection efficiencies.
Based on the excellent Leica CS2 objectives, Leica Microsystems has designed a new objective with optimal chromatic correction and transmission for STED 3X. The Leica HC PL APO 100x/1.40 OIL STED WHITE enables you to perform STED microscopy in the full spectrum of visible light.