ZEISS Lattice SIM 3

Higher imaging speeds and decreased light exposures are a constant demand in imaging experiments. The robustness and flexibility of ZEISS Lattice SIM 3 structured illumination patterns plus the image reconstruction software allow a significant reduction to the number of phase images required for SIM Apotome acquisition mode, and, importantly, this only causes a slight decrease in the resolution of the final images. SIM Apotome acquisition can be operated at 3 phase images per frame, increasing the imaging speed by 66 %. The increased speed is also advantageous for fast screening of large sample areas, such as tissue sections.

In combination with the Leap mode, you can further decrease the number of phase images per final frame to enable the gentlest super-resolution imaging possible.

Burst mode processing uses the rolling window approach to let you observe processes in your living samples at up to 255 fps. Since Burst mode is a post-acquisition step, you have the flexibility to use it with previously acquired data sets. You decide how much temporal resolution is required for your data analysis.

For demanding fast imaging in 3D, the Leap mode acquisition enables you to reduce your imaging time and lower the light exposure on your sample. This works by imaging only every third plane, for three-times higher volume imaging speed and three-times fewer light exposures.

Immunofluorescence of tissue sections is commonly used in immunological research to investigate distribution of and interactions between pathogens and immune cells, all with the aim to develop novel therapies for pathogenic diseases. For compelling results, it is not only crucial to image complete sections as to not miss relevant areas but also to image with enough resolution to identify and quantify individual events.

In the application example shown here, skin tissue sections were imaged to investigate distribution of CD8 cells relative to Leishmania parasite infection sites. The enlarged area is a digital zoom-in only, meaning that it is possible to zoom into any region of the overview image and quantify cell nuclei, CD8 cells and Leishmania parasites.

The synaptic structure and in particular the actives zones where synaptic vesicles are released are key players in signal transmission and proper function of neurons. The imaging of active zones require resolution beyond what can be achieved by standard confocal microscopy.

The lab of Prof. Sean Sweeney investigates a novel mutant that is a regulator of neuronal survival and metabolic responses. Nervous system and synapses are co-labelled with synaptotagmins to observe the general structure of the synapse and distribution of the presynaptic vesicles. Super-resolution microscopy helps identify and quantify the differences in synapse structure and composition of the active zones.

Live yeast cells are among the most challenging samples in fluorescence microscopy. They are extremely light sensitive and smaller than most used cell lines. Furthermore, yeast cells grow in suspension; they can move freely in the culture dish and are of spherical shape, without clearly defined orientation. Tackling all these challenges requires extremely gentle and fast imaging combined with high resolution in all spatial dimensions.

SIM² Apotome is the perfect tool to image live yeast cells with super-resolution, yet fast and gentle enough to observe the cells over extended periods of time. The example clearly demonstrates this unique capabilities. Various subcellular compartments (surface marker, endosomes, vacuole, endoplasmic reticulum) were tagged with superfolder GFP and imaged for 12 hours.

SIM Apotome combined with reduced phases and Leap mode allows you to image large volumes extremely fast and efficiently. Crunch through large volumes fast by recording only one raw image per final reconstructed image. Select regions of interest, switch objectives and use Lattice SIM to gain super-resolution images with a lateral resolution down to 140 nm within the context of the whole sample.

A novel clearing and embedding technology developed by Prof. Tang and his team (Hsiao et al., Nature Communications 2023) combined with the advantages of SIM Apotome acquisition and excellent image reconstruction technology enabled us to image an entire mouse intestine section of 3 mm × 4 mm and ~200 µm thickness within a couple of minutes. Networks of blood vessels and nerves can be visualized with finest details even at depth.