Invitrogen™ EVOS™ S1000 Spatial Imaging System

The EVOS S1000 system features a flexible hardware configuration that includes a custom LED-based light engine and multiple dichroic and emission filters, as well as a high sensitivity sCMOS 4.2-megapixel camera (6.5-μm pixel size) to detect even the most intricate tissue architecture. The system accommodates up to four slides and includes three pre-set objectives (2.5X ,10X, and 20X) with the option of adding other apochromatic-type 5X and 40X objectives.

The instrument is controlled by the EVOS S1000 spatial software, which provides a sophisticated yet simple interface for acquisition of whole tissue scans with integrated spectral unmixing. The software offers fast overview scans in both transmitted brightfield and fluorescence modes and 'periscope' mode for live tissue navigation. It also provides a sophisticated autofocus functionality to enable stitching and of whole slide tissue scans, all seamlessly integrated to avoid any unnecessary post-acquisition, offline processing steps.

The EVOS S1000 Spatial Imaging System offers these important advantages:

• User-friendly protocol builder that allows easy selection of fluorophores from a dropdown menu to automatically set the optical configuration needed by the instrument to capture the desired plexity
• Intuitive auto-expose and powerful auto-focus (laser-based) functions to promptly locate and identify the targets of interest
• Straightforward and consistent guided workflow for spectrally resolving (via linear unmixing) up to 9-plex samples, using unstained and single-color controls
• Off-the-shelf and ready-to-use protocols for 9-plex and 7-plex imaging, pre-created from reference tissues, that can be used to start imaging archived tissue samples with minimal effort
• Noticeably fast 9-plex acquisition of customizable area scans or whole slides with unmixing and stitching performed automatically, as part of the imaging acquisition workflow
• Output of high-quality, OME-TIFF images (with a resolution of 325 nm/pixel at 20X) that are already stitched, spectrally resolved, and ready to use with any third-party software for downstream analysis, eliminating the need for intermediate adjustments
• Full compatibility with fluorescent dyes from multiple vendors, including Alexa Fluor, Alexa Fluor Plus, Aluorä Spatial Amplification, and Opalä dyes, allowing the use of either primary antibody fluorescence conjugates or spatial amplification labeling methods
• Full compatibility with standard tissue slides and coverslips and no proprietary consumables needed
• High sensitivity sCMOS camera (2040 x 2040 pixel resolution, 4.2 Megapixels) with 6.5‐μm pixel size

Spectral imaging for spatial biology, made simple

The EVOS S1000 Spatial Imaging System takes advantage of the power of spectral imaging to simultaneously resolve 9 fluorophores (8 protein targets and DAPI) in a single acquisition round. The system is designed to allow dye selection from a choice of more than 30 different fluorophores with emission spectra between 450 and 810 nm. It uses built-in spectral separation algorithms (based upon linear unmixing), allowing protein targets to be clearly and confidently resolved without the need for iterative labeling and imaging methods, helping preserve valuable tissue samples.

The flexible spectral technology provides possibilities to apply and even combine the tissue labeling methods of your choice to successfully detect targets with different expression levels. All the steps of the acquisition workflow, leading to a high quality 9-plex image, are designed for ease of understanding, even by users who may be new to spatial imaging and spectral unmixing.

Future-proof and dependable hardware

The EVOS S1000 Spatial Imaging System has a 4-position precision slide holder on a motorized, software-controlled, automated stage. The instrument can spectrally detect more than 60 distinct excitation and emission wavelength combinations, ensuring that no additional filters are required for future spatial proteomics imaging projects. With a 5-position objective capacity, magnifications can be customized between 2.5X and 40X with apochromat-type objectives.

A high sensitivity sCMOS camera (2040 x 2040 pixel resolution) is used to capture 16-bit monochrome OME-TIFF images. Generated images can be saved on the internal hard drive (2X 8-TB SSDs for data), an external USB device, or customer IT-approved cloud solutions and local networking. An external Dell™ XE4 PC computer with 12th generation Intel™ Core™ i9-12900 processor, 128 GB DDR4 RAM, and NVIDIA™ Quadro RTX™ A4000 graphics card is used to operate the EVOS S1000 Spatial Imaging System.

Powerful software designed for sophisticated simplicity

The EVOS S1000 Spatial Software presents users with the opportunity to access multiplex imaging on tissue in the simplest possible way. The acquisition interface has been designed with a strong emphasis on usability, even for inexperienced users and those new to spectral unmixing. It starts with an extremely fast overview scan that can be performed either in transmitted or fluorescent mode. For example, four slides can be scanned in fluorescent mode, including the labels, in ~180 seconds.

Live ('periscope') mode is available for quickly navigating the tissue. Finding samples is fast and precise, using a powerful combination of custom IR laser focus and software autofocus. A guided workflow allows for the unmixing matrix creation process to be performed efficiently and confidently, including the creation of an unmixing quality metrics report with qualitative and quantitative parameters to evaluate the quality of the generated spectral unmixing matrix.

The EVOS S1000 Spatial Software can be used to run variable magnification scans of FOV and ROI with multiple options for automated routines. Whole slide stitching and unmixing is conducted automatically as part of the imaging acquisition.

Readiness for downstream analysis

Images acquired on the EVOS S1000 Spatial Imaging System can be further analyzed using any third-party software available that supports pyramidal OME-TIFF file output for handling of large multidimensional image data. Examples are the Halo software by Indica Labs and the freely available open-source QuPath analysis software.

The EVOS S1000 Spatial Imaging System has been built from the ground up to make spatial biology, and particularly proteomics tissue imaging, available to any lab.