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Generate high-resolution 9-plex tissue images in hours
The EVOS S1000 Spatial Imaging System redefines tissue imaging with powerful multiplexing capabilities—enabling integrated acquisition, processing, and high-resolution image generation for spatial localization of proteins with a simplified and fast workflow. Well-suited for researchers who want to quickly image up to 9 markers in a single round, the EVOS S1000 Spatial Imaging System visualizes complex cellular neighborhoods, interactions and spatial relationships with ease.
Why EVOS S1000 Spatial Imaging System?
High-resolution multiplex imaging
Image up to 9 targets simultaneously to reduce workflow complexity while preserving the integrity of valuable tissue.
Automated spectral unmixing
Reduce fluorescence crosstalk, including autofluorescence, and improve signal separation for cleaner multiplex data with integrated spectral unmixing.
Faster time-to-results
Acquire whole slide multiplex tissue images in hours with a streamlined workflow that includes seamless stitching.
Standardized, ready-to-use outputs
Generate spectrally unmixed images as standardized OME-TIFF files that ensure workflow compatibility with any third-party analysis software, including AI pipelines.
Flexible labeling workflow
Use verified Invitrogen reagents for flexible target detection.
Explore the plexibilities with EVOS S1000
More colors. More context. More insights.
The EVOS S1000 Spatial Imaging System transforms multiplex immunofluorescence (mIF) by making higher plex spatial imaging accessible, efficient, and cost-effective (Table 1). With this platform, it is possible to capture, visualize, and generate more biological information from every tissue section in a single imaging run (Figure 1, Figure 2), including:
- Cell types, cell states and cell functions
- Cellular identity in time and space
- Cell to cell interactions
- Cellular “neighborhoods” or networks
- Tissue microenvironments and architecture in their native state
By simultaneously capturing and resolving up to eight fluorescence targets plus nuclear stain (9-plex), the EVOS S1000 Spatial Imaging System generates significantly more data for downstream analysis in a single run, compared to conventional imaging (1–4 colors), and at a fraction of the cost of cyclic imaging.
Table 1. Benefits of multiplex immunofluorescence staining.
|
Single stain |
2 to 4 colors |
Multiplex (≥5 colors) |
Localization of protein |
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Colocalization of proteins |
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|
|
Biomarker expression |
|
|
|
RNA and protein detection in one sample |
|
|
|
Complex cell phenotypes |
|
|
|
Tissue structure |
|
|
|
Characterization of cellular neighborhoods |
|
|
|
Figure 1. Detecting more targets helps provide more details about the tissue microenvironment and highlights the complexity of biological systems within tissues. Images of normal colon (left) and adenocarcinoma tissues (right) stained with the 9-plex colon panel on the EVOS S1000 Spatial Imaging System. Multiplex immunofluorescence staining enables information to be collected about the localization and interaction of biomolecules and cells within the tissue microenvironment.
Figure 2. Multiplex capabilities allow the visualization of more targets in every sample. Unmixed multifield region of an axial Murine kidney FFPE sample labeled with 8 Aluora dyes targeting aquaporins 1, 2, and 4, cytokeratins 8, 18, and 19, MCM2, and smooth muscle actin (SMA) and counterstained with DAPI. Panels 1–4 show the same area of interest and illustrate that increasing the number of labeled targets within a sample produces greater detail. The images were captured using the 20x objective on the Invitrogen EVOS S1000 Spatial Imaging System.
Figure 3. Stained invasive ductal carcinoma shown in individual tiles. Human invasive ductal carcinoma of breast tissue processed and stained with DAB or hematoxylin. Tiles represent individual targets. Images were taken on the EVOS S1000 Spatial Imaging System.
Figure 4. Invasive ductal carcinoma tissue stained with the 8-plex Aluora spatial amplification assay and DAPI. Human invasive ductal carcinoma of breast tissue processed and stained with the Aluora Spatial Rainbow Kit (Cat. No. A40002450). Images and spectral unmixing were performed on the EVOS S1000 Spatial Imaging System. Data was analyzed for single cell segmentation and phenotyping to reveal spatial distribution of immune cell subpopulations. Analysis of the multiplex immunofluorescence stitched image was performed on the Indica Labs HALO (version 4.0.5107.318) software.
Features
With the EVOS S1000 Spatial Imaging System you can:
- Perform spectral fluorescence, transmitted brightfield, phase-contrast, and color brightfield imaging with customizable magnifications (2.5–40x).
- Scan, spectrally unmix, stitch and save a 9-plex fluorescence image of a 1 cm2 tissue area at 20x in approximately 45 min
- Experience live tissue exploration, protocol set-up, spectral extraction, image collection and data retrieval in the simplest possible way, for users with different levels of experience.
- Generate TIFF or fully standardized OME-TIFF files with either single or pyramidal structure, so you can use the third-party analysis software of your choice, such as HaLo and QuPath.
- Select and utilize your preferred antibodies and reagents, as the EVOS S1000 system is compatible with multiple labeling technologies.
Spectral unmixing technology
The EVOS S1000 Spatial Imaging System software facilitates spectral unmixing automatically whenever needed. Spectral unmixing is a process used to resolve signals from fluorophores with significant spectral overlap. This enables higher multiplexing than conventional fluorescence microscopy by eliminating the need for spectrally distinct fluorophores and expanding the number of targets that can be resolved in a single imaging acquisition round (Figure 5). Additionally, unmixing can help resolve tissue autofluorescence from fluorophores.
Perform imaging faster
Complete a full round of 9-plex mIF faster than traditional cyclic technologies to accelerate your experiments.
Reduce the chance for tissue damage
Iterative staining using bleaching or antibody-removal methods extends experimental time and poses risks of epitope loss, tissue degradation, and incomplete fluorophore inactivation across multiple cycles.
Autofluorescence removal
Enhance your imaging precision and improve the accuracy of your cell detection.
Gain confidence in your experiments
Find peace of mind for imaging-stained multiplex tissue samples with software that automatically generates and applies an unmixing matrix to spectrally mixed multiplex tissue samples, clearly identifying fluorophore emissions that bleed into neighboring channels.
See spectral unmixing in action by sliding the toggle between Raw and Unmixed.
This unmixing process leverages unique spectral signatures from each fluorophore to determine the abundance of different signal inputs at each pixel, allowing for precise identification and mapping of various fluorophores within an imaged sample (Figure 6). For the algorithm to function effectively, reference spectra are required to extract the spectrum of each fluorophore, which can be obtained using either predicted spectra (default) or measured spectra through careful preparation of single-color controls. Additionally, an unstained sample is necessary to define the tissue's autofluorescence, which is extracted as an independent spectral signature alongside the fluorophores used in the experiment.
Once all these components are collected, the EVOS S1000 software generates and saves the unmixing matrix to the imaging protocol (Figure 7).
Figure 6. The EVOS S1000 Spatial Imaging System allows capturing multiplex immunofluorescence images through its spectral unmixing capabilities. These spectra show emissions of eight Alexa Fluor and Alexa Fluor Plus dyes and DAPI. Despite the overlap, the built-in algorithms in the EVOS S1000 spatial imaging software can determine the relative contribution of each fluorophore to every pixel of the image and eliminate the spectral bleedthrough from overlapping channels.
Figure 7. The EVOS S1000 Spatial Imaging System generates multiplexed data through its spectral unmixing capabilities. This software feature allows researchers to easily visualize all channels simultaneously and provides a quality metrics report to facilitate highly resolved data.
An Unmixing Quality Metric Report is generated before time is spent imaging full tissue scans by enabling a priori evaluation of the experimental panel (Figure 8). This report provides guidance and metrics demonstrating that the bleedthrough from spectrally overlapping markers will be removed after the unmixing protocol is applied to spectrally mixed multiplex tissue samples.
Figure 8. Visualization of the raw images (left) and unmixed images (right) for each single-color control sample, displayed in each column, across the primary channels shown in each row.
Autofluorescence removal
The EVOS S1000 Spatial Imaging System software facilitates the spectral unmixing of tissue autofluorescence, effectively separating highly autofluorescent signals from desired fluorophore staining (Figure 9). Tissue autofluorescence is present in all tissues to varying degrees and is often strongest in channels where fluorophores such as DAPI are imaged. If left in the image data, autofluorescence can interfere with downstream analysis.
See autofluorescence removal by spectral unmixing in action by sliding the toggle between Raw and Unmixed.
Without autofluorescence removal, downstream image analysis becomes more challenging due to uncertainty about whether pixel intensities belong to a desired marker, such as DAPI, which is commonly used for nuclear segmentation, or tissue autofluorescence. By removing the contribution of tissue autofluorescence from DAPI staining, a much higher confidence cell segmentation mask can be obtained using various nuclear segmentation methods (Figure 10).
Figure 10. Cell detection performed on DAPI without unmixing (top) compared to cell detection performed on DAPI after unmixing and autofluorescence removal (bottom).
Simplified tissue imaging workflow
Unlike existing technologies that can take several days to weeks, the EVOS S1000 Spatial Imaging System can complete the imaging process within several hours with the capability to generate a fully stitched, unmixed, multiplexed image for multiple samples. The system offers flexibility to use a variety of labeling technologies, allowing you to select and utilize your preferred antibodies and reagents.
mIF experimental workflow
To build a panel for the EVOS S1000 Spatial Imaging System, choose your targets, ensuring they align with your research goals. Next, create the panel by carefully selecting antibodies. Use the Invitrogen SpectraViewer to determine where to place the 8 fluorophores labeling the antibodies plus DAPI. When selecting a staining and labeling technology, use the Spatial Biology Reagent Selection Tool. Finally, proceed with the imaging step to capture and analyze your samples.
Ordering information
Every EVOS S1000 Spatial Imaging System includes the imaging unit as well as the objectives (1 of each type), slide holders (4), calibration slide, and AB Assurance support plan listed below.
Discover what pioneering scientists are saying
Customer story: SciLifeLab, Stockholm, Sweden, Spatial Proteomics Unit
Using the Invitrogen EVOS S1000 Spatial Imaging System simplifies the tissue imaging process and provides multiplex data in less than one hour.
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