Multicolor Image Gallery

Multicolor images taken with Leica Microsystems instruments

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Fluorescence multicolor microscopy, which is one aspect of multiplex imaging, allows for the observation and analysis of multiple elements within the same sample – each tagged with a different fluorescent dye – and the same experiment. This results in efficient experiments but also in more reliable and meaningful results to understand complex processes within cells and tissues. This gallery shows images from specimens tagged with multiple fluorescent probes obtained with THUNDER and the STELLARIS platform.

Multicolor Imaging with STELLARIS Confocal Platform

The STELLARIS family of confocal microscopes is equipped with next generation white light lasers (WLL). Paired with Power HyD detectors, STELLARIS provides complete spectral freedom. You can perform multicolor experiments to fit your research needs without being limited by the instrument. STELLARIS can also add an additional dimension to multicolor imaging by using the lifetime-based properties of fluorescent dyes/proteins with the TauSense set of tools.

Root-hypocotyl junction of Arabidopsis thaliana

Nature is multi-dimensional. To truly comprehend its complexity, we need to look in multiple dimensions. This detailed multicolor image of Arabadopsis thaliana root junction was obtained with just one click and one detector. With conventional confocal microscopy, it would be black and white. With STELLARIS what we see is transformed by an extra dimension of lifetime information. Not only is the remarkable architectural diversity of the actin-containing root system revealed with more clarity, but also its relationship to the cell wall and chloroplasts—structures that would be difficult or impossible to distinguish using the intensity data alone. The result is not just a snapshot of nature’s beauty, but an information-rich image that progresses our understanding of plant biology.

Mitotic COS7 mitotic cells - STELLARIS

Uncovering hidden connections between cancer and mitosis. Unravelling the role of mitotic instability in a complex disease like cancer requires correlation of multiple biological markers in the same sample. This image faithfully captures individual COS-7 cells at three stages of mitosis: metaphase, late anaphase and telophase. The delicate interplay between chromosomes, spindle fibers, Golgi, mitochondria and actin cortex is revealed in exceptional detail. The power of STELLARIS to distinguish fluorescent labels that have closely overlapping spectra was essential for this 5-color experiment, taking multiplexing capability beyond ordinary limits.

Cytoskeletal and mitochondrial interactions revealed - STELLARIS

Most conventional confocal systems are equipped with only one or two discrete laser lines in the red. In addition, conventional GaAsP detectors are not sensitive in the NIR range. Both of these factors severely limit the number of red dyes that can be imaged in a single experiment with traditional confocal imaging systems. In the experiment shown here, Cos-7 cells were labeled with red-excitable probes specific for actin, mitochondrial outer membrane, and tubulin. Traditional confocal microscopy with a GaAsP detector detects only the SiRActin (Left). Using the extended WLL on STELLARIS 8 together with the HyD R detector, all 3 fluorescent probes were optimally excited and are visible in the image (Right). Imaging on STELLARIS not only made it possible to see the delicate interplay between mitochondria, actin and tubulin, but freed up the rest of the spectrum for additional fluorescent probes.

Getting the most from live-cell experiments: fewer exposures, more information - STELLARIS

When using traditional confocal microscopy to image several different fluorescent labels in the same sample, sequential imaging of each color channel is often needed to avoid spectral bleed-through, which can degrade image quality. In the case of a kinetic experiment, that means you may miss rapid dynamic events due to the increased time it takes to acquire each time point. In addition, your sample remains on the stage for longer, making it more challenging to maintain cell health for the duration the experiment. The image below was captured from live HeLa cells labeled with 4 different fluorophores to identify nuclei, actin, tubulin and plasma membrane. With STELLARIS, it was possible to collect all 4 channels in a single pass, rather than having to image the cells 4 times in succession.

LAS X Navigator – a GPS for your experiment - STELLARIS

Intuitive sample navigation tools are essential to assure optimal setup of complex multicolor experiments and enable efficient exploration of large specimens. The LAS X Navigator is like a GPS for your experiment, helping you to quickly switch from searching image-by-image to seeing a full overview of your sample. With easy access to both the high-level overview and the minute details, the LAS X Navigator ensures that you always have a clear path to high-quality data.

Both the TLD and the TauContrast images were acquired with a 10x objective, and in each case a single tile is shown from the larger stitched image comprised of 201 tiles.

Discover at the speed of LIGHTNING - STELLARIS

Confocal images obtained with STELLARIS contain more information than you see on the first glance. The LIGHTNING detection concept helps to uncover this hidden information from your specimen. With a single click and fully automatically, LIGHTNING extends the resolution of STELLARIS into super-resolution territory: resolve the fine structures and details, as small as 120 nm lateral resolution, which are elsewise simply not visible. Thanks to its deep integration and optimized GPU processing, LIGHTNING allows for the (near) real-time acquisition of super-resolution images with up to five color channels and a large field of view. Due to this flexibility, LIGHTNING works for any type of specimen and experiment.

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