ZEISS Lattice SIM 5​
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ZEISS Lattice SIM 5​

Live Imaging with Uniform Super-Resolution in All Spatial Dimensions​

ZEISS Lattice SIM 5 has been optimized for capturing subcellular structures and their dynamics. Powered by the Lattice SIM technology and the SIM² image reconstruction algorithm, ZEISS Lattice SIM 5 provides you with outstanding super-resolution capabilities down to 60 nm in both living and fixed cells. Additionally, you can choose SIM Apotome imaging mode and a low-magnification objective to achieve fast overview images of your sample before zooming into super-resolution details.​

  • Capture dynamic processes and finest subcellular structures​
  • Optimize to the needs of your living samples​
  • Get more reliable experiment results​

Capture Dynamic Processes and Finest Subcellular Structures​

Equipped with the ZEISS Lattice SIM illumination pattern and the SIM² image reconstruction algorithm, ZEISS Lattice SIM 5 raises structured illumination microscopy (SIM) to a new level. You will always achieve the best possible results, even when using lower light exposures to protect living specimens. Double the conventional SIM resolution and discriminate the finest subcellular structures that are no more than 60 nm apart. The light-efficient Lattice SIM technology provides the gentlest imaging of living and fixed specimens, giving you not only double spatial resolution compared to classic SIM, but also high temporal resolution with up to 255 fps.​

Caption: Actin dynamics in a U2OS cell expressing LifeAct-GFP imaged with the Lattice SIM 3D Leap mode and reduced phases. Objective: Plan-Apochromat 63× / 1.4 Oil

Optimize to the Needs of Living Samples​

The flexibility of ZEISS Lattice SIM 5 allows you to balance the needs of your experiment by prioritizing resolution, speed, or by finding the right balance in between. Use the photon budget to enhance lateral resolution well below 100 nm or reduce the number of required raw images to boost acquisition speed and gentleness. ZEISS Lattice SIM 5 has a number of options for reducing raw images which allows you to select for the best acquisition settings that target your desired spatial and temporal resolution.​

​Caption: Time lapse imaging of the endoplasmic reticulum in a Cos-7 cell reveals highly dynamic structural changes. Sample courtesy of Miyawaki Lab, RIKEN Institute, Japan.​

Cos-7 cells stained for microtubules (anti-tubulin Alexa Fluor 488, cyan) and actin (Phalloidin Alexa Fluor 561, orange)​

Get More Reliable Experiment Results​

Cos-7 cells stained for microtubules (anti-tubulin Alexa Fluor 488, cyan) and actin (Phalloidin Alexa Fluor 561, orange)​

Cos-7 cells stained for microtubules (anti-tubulin Alexa Fluor 488, cyan) and actin (Phalloidin Alexa Fluor 561, orange)​

Get More Reliable Experiment Results​

ZEISS Lattice SIM 5 comes with outstanding out-of-focus light suppression, giving you the sharpest sectioning in widefield microscopy, even for highly scattering samples. The SIM² image reconstruction uses a special SIM point spread function to robustly reconstruct all structured-illumination-based acquisition data of your ZEISS Lattice SIM 5 with minimal image artifacts – for both living and fixed samples. Rest assured knowing that you are basing your experimental conclusions on reproducible data generated from a powerful and proven algorithm.​

​Caption: Cos-7 cells stained for microtubules (anti-tubulin Alexa Fluor 488, cyan) and actin (Phalloidin Alexa Fluor 561, orange)​

The Technology behind ZEISS Lattice SIM 5​

Widefield Lattice SIM
Widefield Lattice SIM
Comparison of widefield and Lattice SIM images of Cos-7 cells stained for actin (Phalloidin Alexa Fluor 568, magenta), microtubules (anti-tubulin Alexa Fluor 488, yellow) and nucleus (Hoechst, blue). Objective: Plan-Apochromat 63× / 1.4 Oil​

Lattice SIM​

Your 3D Super-Resolution Technique for Live Imaging​

In Lattice SIM, the sample is illuminated with a lattice spot pattern instead of grid lines as in conventional SIM. Due to its intrinsic two-dimensionality, the lattice pattern requires only translational repositioning but no rotation. This leads to a dramatic increase in imaging speed. In addition, the lattice pattern provides higher contrast to allow a more robust image reconstruction. Since the sampling efficiency is doubled compared to classic SIM, half as much light exposure is needed making Lattice SIM a preferred live cell imaging technique. ​

Widefield imaging​

The image resolution is physically limited due to the diffraction limit. Additionally, the image quality suffers from out-of-focus blur and background signal.​

Classic SIM imaging​

To generate higher frequencies, the sample is illuminated with a grid pattern and imaged at different rotational and translational positions of this pattern. The processed image has twice the resolution in all three dimensions.​

Lattice SIM imaging​

The sample is illuminated with a lattice spot pattern instead of grid lines. Compared to classic SIM, sampling efficiency is two times higher. The lattice pattern gives higher contrast and is more robust for processing.​

Reconstructed image​

After acquisition, the resulting super-resolution image is calculated. With Lattice SIM, you can image longer with less bleaching and maintain image quality at higher frame rates.​

Images of Cos-7 cell stained with anti-alpha-Tubulin Alexa fluor 488 were processed with the conventional SIM algorithms based on generalized Wiener filter and with the novel SIM² reconstruction. The images show an improvement of resolution for SIM² compared to SIM. Objective: Plan-Apochromat 63× / 1.4 Oil.

SIM² Image Reconstruction​

Double Your SIM Resolution​

SIM² is a groundbreaking image reconstruction algorithm that increases the resolution and sectioning quality of structured illumination microscopy data. SIM² is compatible with all SIM imaging modes and fully integrated in ZEISS ZEN software.​

​Unlike conventional reconstruction algorithms, SIM² is a two-step image reconstruction algorithm. First, order combination, denoising, and frequency suppression filtering are performed. All the effects resulting from these digital image manipulations are translated into a digital SIM point spread function (PSF). The subsequent iterative deconvolution uses this PSF. Similar to the advantages of using experimental PSF for deconvolution of hardware-based microscopy data, the SIM² algorithm is superior to conventional one-step image reconstruction methods in terms of resolution, sectioning, and robustness.​

Widefield SIM Apotome
Widefield SIM Apotome
Comparison of widefield and SIM² Apotome single plane images of U2OS cells stained for actin (phalloidin Alexa Fluor 647, red), microtubules (anti-alpha-tubulin Alexa Fluor 488, green) and nuclei (Hoechst, blue). Objective: LD LCI Plan-Apochromat 25× / 0.8 Imm Corr​

SIM Apotome​

Flexible Optical Sectioning​

Choose the SIM Apotome acquisition mode to achieve fast overview images before zooming into super-resolution details. SIM Apotome uses structured illumination to give you fast optical sectioning of larger volumes with crisp contrast and high resolution in all dimensions.​

SIM Apotome in combination with the SIM² reconstruction algorithm allows you to further adjust the gentleness of fast live-cell imaging with high contrast and resolution. Or use your new optical sectioning speed to increase productivity when acquiring large sample areas or large volumes at different magnifications.​

Boost the Speed of SIM Imaging Even Further​

Increase the temporal resolution and productivity for 2D and 3D imaging by using the speed enhancement modes.​

U2OS cell expressing Rab5-mEmerald (green) and tdTomato tagged Golgi associated transport marker (magenta). Simultaneous dual-color acquisition with an exposure time of 1.5 ms / phase for a FOV of 1024 × 1024 pixel (64 µm × 64 µm). Objective: Plan-Apochromat 63× / 1.4 Oil

2D Burst mode

Get full temporal information​

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.​

U2OS cell expressing calreticulin-tdTomato to visualize the endoplasmic reticulum. The time series shows a maximum intensity projection of the volume data set. Objective: Plan-Apochromat 63× / 1.4 Oil

3D Leap mode

Digital sectioning at a new level​

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. ​

Simultaneous Two-Color Imaging​

Investigation of living samples very often focuses on interactions of different proteins or organelles. Simultaneous imaging of the involved structures is key to proper understanding of these highly dynamic processes. ZEISS Lattice SIM 5 can be equipped with two cameras in parallel and perform true simultaneous two-color imaging within your entire field of view.​

For excellent performance at a cost-efficient price point, select the ZEISS Axiocam 820 mono camera, which features back-illuminated CMOS sensor with a peak quantum efficiency of 86 %. This camera is the perfect choice for imaging faint fluorescence signals in living or fixed samples.​
ZEISS Axiocam 820 mono​
ZEISS Axiocam 820 mono​
For outstanding performance, choose the Hamamatsu ORCA-Fusion BT camera. This camera features a scientific CMOS (sCMOS) with a back-thinned sensor enabling peak quantum efficiency of ~95 %. With its high acquisition speeds and exposure times down to 1 ms, it yields unparallel digital imaging results.​
 Hamamatsu ORCA-Fusion BT ​
Hamamatsu ORCA-Fusion BT ​

Application Examples​

See ZEISS Lattice SIM 5 in Action​

  • Cos-7 cell expressing TOMM20-mEmerald (cyan) and EB3-tdTomato (orange) shows dynamic movement of mitochondria and microtubules. Imaged with Lattice SIM. Objective: Plan-Apochromat 63× / 1.4 Oil​
  • Actin dynamics of U2OS cells expressing LifeAct-tdTomato imaged with the Lattice SIM² mode with reduced phases. Objective: Plan-Apochromat 63× / 1.4 Oil.
  • Actin dynamics in a U2OS cell expressing LifeAct-GFP imaged with the SIM Apotome 3D Leap mode and reduced phases. Objective: Plan-Apochromat 40× / 1.4 Oil​

Balance Your Need for Speed and Resolution​

Higher imaging speeds and decreased light exposures are a constant demand in imaging experiments. The robustness and flexibility of Lattice SIM structured illumination patterns plus the image reconstruction software allow a significant reduction to the number of phase images required for Lattice SIM acquisition mode, and, importantly, this only causes a slight decrease in the resolution of the final images. Lattice SIM acquisition can be operated at 9 phase images per frame instead of 13, increasing the imaging speed by 44 %. The increased speed is particularly advantageous for gentle imaging of highly dynamic live cells where slower acquisition would result in motion blur and reduced resolution.​

​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.

Mouse small intestine. Overview image recorded with SIM Apotome and Plan-Neofluar 10× / 0.3 Air. Region of interest imaged with Lattice SIM and Plan Apochromat 63× / 1.4 Oil. Sample courtesy of Prof. Shiue-Cheng (Tony) Tang, Institute of Biotechnology & Department of Medical Science, National Tsing Hua University, Taiwan​

Resolve the Details Hiding in the Depth​

High-Quality Sectioning and Super-Resolution in Thick Samples​

The Lattice SIM illumination pattern exhibits both higher contrast and deeper sample penetration as compared to classical SIM. Achieve super-resolution images along with high-quality sectioning even in thick or scattering samples.​

​A novel clearing and embedding technology developed by Prof. Tang and his team (Hsiao et al., Nature Communications 2023) combined with the robust Lattice SIM illumination pattern and excellent image reconstruction technology enabled imaging throughout an entire mouse intestine section of ~200 µm thickness. Networks of blood vessels and nerves can be visualized with finest details even at this depth.​

Cos-7 cell expressing Calreticulin-tdTomato (endoplasmic reticulum, magenta) and Tomm20-mEmerald (mitochondria, green) was simultaneously imaged for two colors. The movie shows high dynamic interactions of the ER and mitochondria. Objective: Plan-Apochromat 63× / 1.4 Oil ​

Observe Life’s Finest Details​

Live imaging at high spatiotemporal resolution​

ZEISS Lattice SIM 5 combines high speed imaging with incredible light efficiency, low photon dosage and sensitivity. You can observe cellular, subcellular, and even sub-organelle structures in living specimens in 2D and 3D over time. ​

​Mitochondria are highly dynamic organelles that constantly undergo fusion and fission events to ensure proper distribution of ATP across the cell. In order to do their job, they are known to interact with many other subcellular compartments including microtubules, which they ride on to get to their destinations, or the ER, which wraps around mitochondria to initially constrict their diameter before fission events.​

Downloads

  • ZEISS Lattice SIM 5

    Your live imaging system for uniform super-resolution in all spatial dimensions

    Pages: 23
    File size: 5 MB
  • ZEISS Lattice SIM Family

    Full Access to Super-Resolution Imaging for all Research Areas

    Pages: 6
    File size: 3 MB

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