ZEISS Elyra 7 with Lattice SIM

Your Flexible Platform for Fast and Gentle 3D Superresolution Microscopy

Your life sciences research often requires you to measure, quantify and understand the finest details and sub-cellular structures of your sample. You may be working with tissue, bacteria, organoids, neurons, living or fixed -cells and many different labels.

Elyra 7 with Lattice SIM takes you beyond the diffraction limit of conventional microscopy to image your samples with superresolution. You examine the fastest processes in living samples – in large fields of view, in 3D, over long time periods, and with multiple colors. The new Lattice SIM technology of your Elyra 7 brings structured illumination microscopy (SIM) to a new level. Groundbreaking light efficiency gives you gentle superresolution imaging with incredibly high speed – at 255 fps you will get your data faster than ever before.

Elyra 7 lets you combine Lattice SIM with single molecule localization microscopy (SMLM) for techniques such as PALM, dSTORM and PAINT. You can now choose freely among your labels when imaging with resolutions down to 20 nm laterally. High power laser lines allow you to image your sample with ease, from green to far red.

Elyra 7 is also very flexible: you can employ a wealth of contrasting techniques and combine them with optical sectioning. The new Apotome mode gives you superfast optical sectioning of your 3D samples. All that, plus Elyra 7 works seamlessly with your ZEISS SEMs in a correlative workflow.

Highlights

Lattice SIM – Superfast and Gentle Superresolution Microscopy

You can now use the novel Lattice SIM to uncover new mechanistic details and quantify the finest subcellular structures in large fields of view. This is a real breakthrough in light efficiency, enabling fast and gentle superresolution imaging of living specimens. Elyra 7 excels even more when it comes to fast imaging of 3D volumes at excellent z-resolution. Whether in 2D or 3D, by illuminating your samples with lower laser exposure, you minimize photodamage and so can observe fast cellular processes such as vesicle trafficking, membrane ruffling and signaling.  

Lattice SIM: U2Os cell expressing an mEmerald-GFP tagged endosomal transport marker (Rab5a) and tdTomato tagged golgi and golgi associated transport marker.

Optimized Localization Microscopy

Single molecule localization microscopy (SMLM) gives you access to molecular mechanisms in both fixed and living specimens. You can count molecules and come to understand, molecule-by- molecule, how individual proteins are arranged within a structural context. Elyra 7's SMLM module delivers molecular resolution in large 3D volumes and powerful post-processing algorithms for quantification. With its efficient dual camera detection and high power laser lines across the visible spectrum, you're free to choose your dyes and markers for your experiments.

Xenopus laevis A6 cells (epithelial kidney cells), imaged with SMLM. Gp120, a nuclear pore complex protein arranged with eightfold symmetry was labeled with Alexa Fluor 647

Freedom for Your Experiments

Elyra 7 allows you to choose and combine the best imaging techniques for your experiments, now and in the future. Select the modules you need today – Lattice SIM, SMLM or a combination of both – then expand your system later, as your needs grow. Elyra 7 is not just a superb superresolution microscope: it's your flexible platform for live cell imaging. You can match the spatial and temporal resolution perfectly to your applications. Upgrade your system anytime with a whole range of additional options. Or use ZEN imaging software and correlative microscopy workflows to combine your data with complementary imaging modalities.

ZEISS ZEN Shuttle & Find with Elyra 7 and GeminiSEM


ZEISS ZEN Shuttle & Find with Elyra 7 and GeminiSEM

The Technology Behind It

Lattice SIM

Elyra 7's light-efficient Lattice SIM illumination pushes the boundaries of fast superresolution acquisition with minimized impact on the specimen. Lattice SIM provides optical sectioning and a doubling of diffraction-limited resolution in 3D (120 nm in xy and 300 nm in z). Elyra 7 provides optimal image quality and resolution across the entire visible spectrum with a large field-of-view. Lattice SIM gives you even more possibilities for increasing your image acquisition speed. Accelerate your acquisition of volumes by a factor of three, or push your 2D frame rate even further – up to 255 fps. You can precisely match the achievable spatial resolution and frame rate of Elyra 7 with all your scientific needs. Lattice SIM leaves you free to image faster and longer than ever before – without compromising on resolution.

Capture Fast Dynamics

Lattice SIM lets you observe superresolution processes with unprecedented speed.

Lattice SIM: U2Os cell expressing an mEmerald-GFP tagged endosomal transport marker (Rab5a) and tdTomato tagged golgi and golgi associated transport marker. The resulting images were acquired with a frame rate of >200 fps, allowing the detection of rapid events.

Gentle Superresolution Imaging

Reduce the light dosage on your specimen and still capture all the details – in multiple colors.

Lattice SIM: Tomm20-mEmerald and EB3-tdTomato in a U2Os cell were imaged simultaneously for more than 1400 frames.

Resolve the Finest Details

Achieve optimal resolution across all wavelengths with multiple objectives.

Lattice SIM: Synaptonemal complex from mouse testis, spread on a coverslip. Sycp1 is labeled with Alexa Fluor 488 (green) and Sycp3 is labeled with Alexa Fluor 568 (magenta). Sample: courtesy of M. Spindler and R. Benavente, University of Würzburg, Germany. 

How Lattice SIM works

In classic SIM, the sample area is illuminated and imaged with changing grid line direction and position. The grid structures interfere with sample structures, creating Moire fringes. These contain high frequency information – that is, high resolution information – transformed down to low frequencies that can be resolved by the optical system. After acquisition, the resulting image will have twice the resolution in all three dimensions and can be computed.

In Lattice SIM, the sample area is illuminated with a lattice spot pattern instead of grid lines. The lattice pattern gives higher contrast and is more robust for processing. Sampling efficiency is 2× higher than with classic SIM. As a result, you need less illumination.

It’s up to you how you use this improved photon efficiency. You can image faster with high image quality and low bleaching. Or get better image quality at the same speed and low bleaching. Or image more gently with high speed and image quality. It’s your choice.

Watch the movie for a quick comparison of classic SIM and Lattice SIM

Single Molecule Localization Microscopy

Single-molecule localization microscopy (SMLM) encompasses techniques such as PALM, dSTORM, and PAINT. With high power lasers across the visible spectrum and dual camera detection, Elyra 7 allows researchers to gain access to a broad range of dyes, markers and fluorophores in almost any possible combination. Elyra 7 enables quantification with consistent precision over a large field-of-view and an unprecedented z-capture range. It allows 3D acquisition of a whole cell with molecular precision.

Resolve Molecular Structures

SMLM allows you to map precise locations of individual proteins.
 

SMLM: Eightfold symmetry of the nuclear pore complex in A6 cell.
SMLM: Eightfold symmetry of the nuclear pore complex in A6 cell.

Determine the Relationships Between Molecules

Detect two channels with molecular precision.
 

SMLM: Alpha tubulin was labelled with Alexa 555 and beta tubulin with Alexa 488.
SMLM: Alpha tubulin was labelled with Alexa 555 and beta tubulin with Alexa 488.

Capture Information in Three Dimensions

Untangle molecular relationships in z with confidence.

SMLM: With Elyra 7 you can image a z-depth of 1.4 µm in a single acquisition.
SMLM: With Elyra 7 you can image a z-depth of 1.4 µm in a single acquisition.

How SMLM works

In SMLM, photo-switchable fluorescent molecules are sparsely activated so that only one out of many will be in its on-state within a single point spread function (PSF). This lets you determine its center of mass with a localization precision that far exceeds the extension of the PSF. Once recorded the molecule is turned to its off-state – for example, by photobleaching – and the cycle of activation / deactivation is repeated again and again until all molecules are captured.

The localizations are plotted in a new image to create the super-resolved image. If the PSF shape codes for the z-position, the method works in 3D as well. Expect to achieve resolutions in the range of 20 – 30 nm laterally and 50 – 80 nm axially.

With Elyra 7, powerful laser lines across the visible spectrum give you freedom to choose the best dyes for your experiments. Plus, the dual camera option with precise synchronization allows you to capture two labels simultaneously.

The basic principle of PALM. Move the slider to see how this technology works.

Get superfast optical sectioning with the new Apotome mode

You know the challenge: live cell imaging with a widefield system often suffers from out-of-focus blur or background signal. These effects can decrease contrast and resolution of your images. The new Apotome mode of your Elyra 7 now uses structured illumination to give you fast optical sectioning with crisp contrast and high lateral and axial resolution.

How the Apotome mode works

A grid pattern is used to illuminate and rapidly modulate the fluorescence signals in the focal plane of your microscope. After acquiring five images with different grid positions, ZEN imaging software combines these frames into a resulting image which contains only information from the focal plane – your optical section. The new Apotome mode now allows you to perform fast and gentle live cell imaging with high contrast and resolution.

Or, you can use your new optical sectioning speed to increase your productivity when acquiring large sample areas or big volumes.

Penicillium autofluorescence. The Apotome mode allowed to image a volume of 90 × 90 × 50 µm with 422 Z-planes.

COS-7 cells. Maximum intensity projection of 66 sections.

COS-7 cells. Maximum intensity projection of 66 sections. Microtubules stained with Alexa 488 (green) and Actin stained with Alexa 568 (red). The Apotome mode allowed simultaneous dual color acquisition.

Application Examples

Lattice SIM: Observe cellular processes over long time periods without perturbing your specimen. U2Os cell expressing an mEmerald-GFP tagged endosomal transport marker (Rab5a) and tdTomato tagged golgi and golgi associated transport marker. Simultaneous dual-color acquisition over a period of 30 minutes.

Lattice SIM: Put high resolution details in the context of a whole cell. Cos7 cell expressing EB3-tdTomato. Sample courtesy of M. Sauer, University of Würzburg, Germany.

Lattice SIM: 3D volume image of Thy1-GFP neurons in a mouse brain section. A ~20 µm z-stack was acquired inside of the tissue section. Sample courtesy of Herms lab, DZNE, Munich, Germany.

Lattice SIM: Resolve fast dynamics without photobleaching. U2Os cell expressing Lifeact-9 (labelling actin) and EB3-mEmer-ald-GFP (labeling growing ends of microtubules). Sequence of 100 images taken simultaneously in 2 colors. The motion of EB3 and Lifeact is followed with little appreciable photobleaching over the course of several minutes.

Widefield image
Lattice SIM image

Observe the finest details. Actin labeled with Phalloidi, showing the two fold resolution improvement of Lattice SIM.

Lattice SIM: 3D image of microtubules, color coded for depth.

Lattice SIM: 3D image of microtubules, color coded for depth.

Lattice SIM: Observe cellular processes over long time periods without perturbing your specimen. U2Os cell expressing an mEmerald-GFP tagged endosomal transport marker (Rab5a) and tdTomato tagged golgi and golgi associated transport marker. Simultaneous dual-color acquisition over a period of 30 minutes.

Lattice SIM: Put high resolution details in the context of a whole cell. Cos7 cell expressing EB3-tdTomato. Sample courtesy of M. Sauer, University of Würzburg, Germany.

SMLM: 3D PAINT image of mitochondrial membranes in BSC1 (kidney epithelial cells)

SMLM: 3D PAINT image of mitochondrial membranes in BSC1 (kidney epithelial cells). The outer membrane protein TOM 20 was labeled using Ultivue – I2-650 imaging strand. Widefield image.

SMLM: 3D PAINT image of mitochondrial membranes in BSC1 (kidney epithelial cells).

SMLM: 3D PAINT image of mitochondrial membranes in BSC1 (kidney epithelial cells). The outer membrane protein TOM 20 was labeled using Ultivue – I2-650 imaging strand. 3D PAINT image color coded for z-depth.

SMLM: 3D PAINT image of mitochondrial membranes in BSC1 (kidney epithelial cells). The outer membrane protein TOM 20 was labeled using Ultivue – I2-650 imaging strand. Individual z-plane showing mitochondrial membrane structure.

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ZEISS Elyra 7

Your Flexible Platform with Lattice SIM for Fast and Gentle 3D Superresolution

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Introducing Lattice SIM for ZEISS Elyra 7

Structured Illumination Microscopy with a 3D Lattice for Live Cell Imaging

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