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EVOS S1000空间生物学成像分析系统简化了蛋白质空间定位的组织成像流程。
Invitrogen EVOS S1000空间生物学成像分析系统是一款宽场光谱成像系统，整合了多重光谱荧光、透射明场、相差及彩色明场成像等诸多功能。
核心特点：
- 快速：20分钟即可完成1 cm²组织的9标成像（10x；扫描、光谱解析及拼接）
- 灵活：兼容多种荧光染料、抗体、RNA探针及显色试剂
- 全面：支持光谱荧光、透射明场、相差及彩色明场成像
- 高灵敏度：配备420万像素16-bit科研级sCMOS相机，可清晰检测组织内复杂的多重染色信号
- 可变倍率：支持2.5x–40x物镜（5孔物镜转盘）
- 高效：自动化激光聚焦与软件辅助聚焦流程加速图像采集
- 易用：直观的软件界面，适用于不同经验水平的用户（图像采集、导航及数据检索）
- 兼容性：支持TIFF、OME-TIFF及分层图像格式，适配各类数据分析软件
- 高分辨率：光谱解析技术减少荧光串色，生成高分辨率图像
图1：使用EVOS S1000采集的扁桃体组织染色图像（透射明场图与彩色图）。
全面加速空间蛋白质组学研究
观看并了解多重成像技术在空间蛋白质研究中的最新进展。探索EVOS S1000如何简化工作流程、支持多染料应用并实现快速高分辨率成像。了解Invitrogen Aluora空间信号增强试剂如何通过高灵敏度和明亮信号提升抗体灵活性，以及如何利用荧光标记一抗实现快速多重染色。
大家好！我是Thermo Fisher Scientific成像技术高级产品经理（芬兰），另一位是Chris Langsdorf，Thermo Fisher Scientific蛋白质与细胞分析全球产品经理（美国）。
感谢大家参与本次讲座。今天将分享我们在空间蛋白质组学多重成像领域的最新技术工具，帮助您深入理解空间生物学。如需深入了解，欢迎随时联系我们的团队。
讲座议程
1. 空间生物学的革命性意义
2. 多重蛋白质成像的挑战
3. 通过样本标记、成像及分析技术解决上述挑战
空间生物学的优势
通过抗体检测技术，我们不仅能分析单细胞，还能解析其在组织微环境中的时空身份、代谢状态、功能活性及细胞间相互作用。
用户评价
突破传统免疫组化限制
EVOS S1000空间成像系统助力从单一样本中获取更丰富的组织微环境信息。
- Why spatial proteomics?
- Spectral unmixing technology
- Benefits of multiplex IF
- Experimental workflow
为何选择空间蛋白质组学？
通过EVOS S1000的多重免疫荧光成像，结合OME-TIFF格式数据，可捕获以下关键信息：
- 细胞类型、状态与功能
- 细胞时空身份
- 细胞间互作
- 细胞“邻域”或网络
- 原生状态下的组织微环境与结构
图2：结肠癌组织中免疫标记物（α-SMA Alexa Fluor 420、CD6等）与DAPI染色图像（EVOS S1000采集图像）。
图3：左图为图7圈选的结肠腺癌三级淋巴结构（TLS）放大图，右图显示TLS内的细胞身份。
Generate highly resolved multiplex immunofluorescence images in tissue sections
Benefits of EVOS S1000 Imaging System spectral unmixing technology
The EVOS S1000 Spatial Imaging System software facilitates spectral unmixing, a process used to resolve signals from fluorophores with significant spectral overlap, enabling higher multiplexing than conventional fluorescence microscopy by eliminating the need for spectrally distinct fluorophores (Figure 4).
Figure 4: Example of spectral emissions of multiple dyes. This spectra show emissions of 8 Alexa Fluor Plus dyes + DAPI. The emission spectra of multiple dyes overlap and makes it difficult to detect true signals from background noise without unmixing software.
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. 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 5).
Figure 5: 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 enables the evaluation of the experimental panel before time is spent imaging full tissue scans (Figure 6). This report provides guidance and metrics demonstrating that the bleed through from spectrally overlapping markers will be removed after the unmixing protocol is applied to spectrally mixed multiplex tissue samples.
Figure 6. 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.
The resulting data from the EVOS S1000 Spatial Imaging spectral unmixing software provides a high-resolution image (Figure 7).
Figure 7. Human tonsil processed and stained with the Aluora Spatial Rainbow Kit (Cat. No. A40002450) raw (mixed, left) and processed (unmixed, right).
Observe similar staining of structures in the raw channels capturing Aluora 488, Aluora 514, and Aluora 555 dyes, and also Aluora 647 and Aluora 700 dyes due to the overlap of emission spectra. Automated processing (spectral unmixing) of this image with the EVOS S1000 Spatial Imaging software allows for the distinct staining across each channel to be revealed in unmixed images. The resulting composite image is more defined and useful for quantitative imaging. Primary antibodies to each target were detected with Aluora spatial amplification dyes including: Aluora 488 dye with antibody targeting vimentin (blue), Aluora 514 dye with antibody targeting eCadherin (red), Aluora 555 dye with antibody targeting PD1 (orange), Aluora 594 dye with antibody targeting CD31 (cyan), Aluora 647 dye with antibody targeting CD3 (green), Aluora 700 dye with antibody targeting CD163 (magenta), Aluora 750 dye with antibody targeting SMA (white). Tissues were imaged on the EVOS S1000 Spatial Imaging System.
Traditional 1-color staining highlights specific proteins
Figure 8. 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.
Multiplex immunohistochemistry staining highlights the complexity of biological systems within tissues
Figure 9. 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.
Simplified tissue imaging workflow
Unlike existing technologies that can take several days to weeks, the EVOS S1000 instrument can complete the imaging process within several hours, with the greatest time savings found in its capability to generate a fully stitched, unmixed, multiplexed image for multiple samples. With the EVOS S1000 Spatial Imaging System, you have the flexibility to use a variety of labeling technologies, allowing you to select and utilize your preferred antibodies and reagents. The instrument outputs OME-TIFF files that are compatible with any third-party analysis software such as Halo and QuPath (QuPath: Open source software for digial pathology image analysis. Scientific Reports (2017).)
Figure 10: The spatial biology multiplex workflow incorporates the EVOS S1000 Spatial Imaging System that multiplexes more biomarkers with the use of spectral unmixing.
To build a panel for the EVOS S1000 Spatial Imaging system, start by choosing your targets, ensuring they align with your research goals. Next, create the panel by carefully picking out antibodies; use the Invitrogen SpectraViewer to determine where to place the 8 fluorophores labeling the antibodies plus DAPI. For the staining and labeling technology, use our Spatial Biology Reagent Selection Tool to chose what is right for your experiment. Finally, proceed with the imaging step to capture and analyze your samples.
Benefits of spectral unmixing technology
Antibodies and reagents
Ordering information
Every EVOS S1000 Spatial Imaging System (Cat. No. AMFS1000) 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.
Every EVOS S1000 Spatial Imaging System (Cat. No. AMFS1000) 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.
Resources and support
The Invitrogen EVOS S1000 Spatial Imaging System is only available in participating countries and regions including US, Canada, and Europe in 2024.
HALO is a trademark of Indica Labs, Inc.
For Research Use Only. Not for use in diagnostic procedures.