Multicolor Image Gallery

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.

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.

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.

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.

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.