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EVOS M7000 Imaging System
The EVOS M7000 Imaging System offers you these important advantages
The EVOS M7000 Imaging System is a fully automated, minimal personal-contact system designed for user safety that features high-resolution CMOS cameras (mono and color) with improved resolution and sensitivity. Designed with advanced capabilities to simplify demanding cell-based imaging applications such as live-cell analysis, image tiling, and z-stacking, the Invitrogen EVOS M7000 Imaging System brings high performance and fast, automated imaging right to your lab bench. It performs rapid autofocusing, image acquisition and processing of large data.
- Safety—fully automatic, on-screen display, no oculars and many more safety features to prevent cross-contamination
- Speed—scan a 96-well plate in 3 fluorescence channels in less than 5 minutes
- Flexibility—customize the system with more than 20 user-changeable LED light cubes, dual cameras (monochrome and color), a variety of objectives ranging from 1.25x to 100x, and multiple vessel holders
- Time-lapse live-cell imaging—onstage incubator option for precise control of temperature, humidity, and gases for normoxic or hypoxic conditions allows a wide range of biological studies under physiological conditions
- Two cameras, no compromises—all systems come with two cameras: a dedicated high-sensitivity monochrome camera optimized for fluorescence imaging and quantitation, and a dedicated high-resolution color camera optimized for colorimetric imaging
- Area view—move rapidly and seamlessly between single-field mode and low- and high-magnification scan modes to easily define and capture the area of interest
- Automation—time-saving features such as autofocus, rapid stage movement, and automated routines help reduce time to complete experiments, allowing high throughput, high data quality, and improved experimental reproducibility without touching the microscope and without the need for personal contact with the microscope
- Data analysis—extensive quantitative imaging and statistical analysis in combination with Invitrogen Celleste Image Analysis Software, an optional advanced software package offering powerful tools for image segmentation and classification that can be used for cell counting and measuring changes
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EVOS M7000 replaces the EVOS FL Auto 2 and is compatible with the accessories of FL Auto 2, including the EVOS Onstage Incubator for easy and simplified live-cell analysis, image tiling, and z-stacking.
Imaging made more safe with EVOS imaging systems
- On-screen display (no oculars)
- Automated controls, minimal handling
- Swift decontaminations
- Fits in biosafety cabinets
- Easy routine maintenance
Watch and make way for more with Invitrogen EVOS M7000 Imaging System
Powerful, intuitive software
All EVOS microscopes come with onboard software that allows you to view, capture, and process images quickly and easily. Featuring a user interface and workflow designed by biologists for biologists, the software gives you full control over your fluorescence, brightfield, color brightfield, and phase contract images.
On the M7000 and M5000, the onboard software offers pinpoint operational control and powerful image processing tools such as cell counting, confluence, Z-stack visualization, and time-lapse movies. These tools operate intuitively with minimal training, aided by onboard SmartStart guides. Autofocus and other automated tools save time and effort. The M7000 adds image tiling and stitching as well as fully automated capabilities for image acquisition and processing when scanning tissue slices or acquiring data from multiwell plate vessels. The ability to analyze images in batch mode allows you to seamlessly process large quantities of data.
Click image to enlarge
Fast, robust autofocus
The autofocus algorithm on the EVOS M7000 and M5000 is fast and efficient in any channel. For automated routines with the M7000, you can choose the interval at which autofocus is used in an area scan or multiwell plate acquisition.
Click image to enlarge
Cell confluence app
Automatically measure and document the percent confluence of your cultured cells, allowing consistent passaging and increasing reproducibility of downstream cell-based assays. Use it for a single field or in batch mode for multiple fields and wells.
Click image to enlarge
Automatic cell counting
Advanced tools let you identify and count fluorescent objects while gating out undesired objects based on size, brightness, and circularity. Apply to a single field or to a multiwell plate using batch analysis.
Click image to enlarge
Measure transfection efficiency in cultured cells
Automatically calculate transfection efficiency based on a user-adjustable threshold of positive pixels in a transmitted light or fluorescence image. Apply to a single field or to multiple fields using batch analysis.
Images acquired with EVOS M7000 and deconvolution performed using Celleste 5.0
Click image to enlarge
Figure 1. U2OS cells labeled with Alexa Fluor Plus 555 Phalloidin and imaged with EVOS M7000. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), Tubulin (Cat. No. 32-2600) detected using Goat anti mouse Alexa Fluor Plus 647 secondary (Cat. No. A32728), and Alexa Fluor Plus 555 Phalloidin (Cat. No. A30106) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 2. HeLa cells labeled with Alexa Fluor Plus 555 Phalloidin and imaged with EVOS M7000. HeLa cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605) and Alexa Fluor Plus 555 Phalloidin (Cat. No. A30106) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650) and RFP (Cat. No. AMEP4652) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 3. HeLa cells labeled with Alexa Fluor Plus 555 Phalloidin and imaged with EVOS M7000. HeLa cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605) and Alexa Fluor Plus 555 Phalloidin (Cat. No. A30106) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650) and RFP (Cat. No. AMEP4652) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 4. HeLa cells labeled with Alexa Fluor Plus 647 Phalloidin and imaged with EVOS M7000. HeLa cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605) and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60 Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650) and Cy5 (Cat. No. AMEP4656) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 5. HeLa cells labeled with Alexa Fluor Plus 647 Phalloidin and imaged with EVOS M7000. HeLa cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605) and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60 Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650) and Cy5 (Cat. No. AMEP4656) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 6. U2OS cells labeled with Alexa Fluor Plus 647 Phalloidin and imaged with EVOS M7000. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), CellLight Mitochondria-RFP BacMam 2.0 (Cat. No. C10601), and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Images acquired with EVOS M7000 and Tissue Stitching
Click image to enlarge
Figure 7. Sagittal Rat brain section detection of neuronal and glial markers and Tissue Stitching in SlowFade Glass. 10 mm cryo-tissue of sagittal section of mouse brain was fixed in PBS buffered 4% paraformaldehyde for 10 min. Afterward the tissue was permeabilized and cleaned in 1% Triton/PBS for 30 minutes. The section was then blocked with 10% normal goat serum followed by additional blocking with ReadyProbes mouse on mouse IgG blocking solution (Cat. No. R37621). Primary antibody of mouse monoclonal against HuC/HuD (Cat. No. A21271) was incubated overnight at 4˚C. Subsequentially, the tissue was labeled using the Alexa Fluor 647, goat anti-mouse secondary HuC/D detection (1000x dilution for 30 min). Tissue was mounted with SlowFade Glass Soft-set Antifade Mountant, with DAPI (Cat. No. S36920), imaging (4x Olympus objective) and tissue stitching was performed on EVOS M7000 Imaging System.
Click image to enlarge
Figure 8. A stitched image showing the live cell density in an entire well of a 6-well plate whole well stitching. Live HeLa cells were stained with calcein AM and imaged with EVOS M7000 using 10x objective. 13x18 stitched image (234 FOV) shows an entire well of a 6-well plate. Note the resolution of the individual FOV is retained when stitching an entire well.
Click image to enlarge
Figure 9. A stitched image showing the fixed cell density in an entire well of a 6-well plate whole well stitching. HeLa cells were fixed with 4% PFA, stained with Eosin-Y and imaged with EVOS M7000 using 10x objective. 13x18 stitched image (234 FOV) shows an entire well of a 6-well plate. Note the resolution of the individual FOV is retained when stitching an entire well.
Click image to enlarge
Figure 10. 384-well spheroid screening. A549 spheroids imaged label-free (brightfield) on an EVOS M7000 using a 10x Olympus SApo objective in a 384-well plate to demonstrate the speed and efficacy of 384-well screening.
Images acquired with EVOS M7000
Click image to enlarge
Figure 11. U2OS cells labeled with Alexa Fluor Plus 555 Phalloidin and imaged with EVOS M7000. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), Tubulin (Cat. No. 32-2600) detected using Goat anti mouse Alexa Fluor Plus 647 secondary (Cat. No. A32728), and Alexa Fluor Plus 555 Phalloidin (Cat. No. A30106) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes.
Click image to enlarge
Figure 12. U2OS cells labeled with Alexa Fluor Plus 647 Phalloidin and imaged with EVOS M7000. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), CellLight Mitochondria-RFP BacMam 2.0 (Cat. No. C10601), and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes.
Olympus 10x Apo objective; transmitted light only; 3x3 tile per well; 3,456 images in 21 mins using EVOS M7000.
DMSO (untreated)
1 µM Jasplakinolide-treated
Brightfield stitch time lapse. U2OS cells monitored label-free over 72 hours, imaged every 30 mins on an EVOS M7000 with an Olympus 10x S-Apo objective in a 37C 5% CO2 incubated environment using an EVOS On-Stage Incubator. Image shown is a 2x2 stitch of DMSO (negative control) treated versus cells treated with 1 µM of the actin-arresting drug Jasplakinolide. While normal cell proliferation can be seen in the control condition, membrane blebbing and internal vesicle transport arrest can be seen in the treated cells due to the arrest of the actin cytoskeleton;
Single FOV
Stitch time lapse
Time-lapse video stitching. Human monocyte-derived macrophages were labeled in a Mattek dish and labeled Ramos added to macrophages along with Hoecsht 33342 and 10nM Rituximab. Red signal indicates phagocytosis and acidification of Ramos cells by macrophages. Images acquired on EVOS M7000 (Cat. No. AMF7000) using a 20x EVOS Fluorite Phase (Cat. No. AMEP4982) air objective. Image shown is a stitched composite of 25 fields of view.
U2OS cells labeled with Alexa Fluor Plus 647 Phalloidin, imaged with EVOS M7000 and deconvolution performed with Celleste 5.0 software. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), CellLight Mitochondria-RFP BacMam 2.0 (Cat. No. C10601), and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes.
Z-stack animation
Z-stack montage
Click image to enlarge
Z-stack animation Z- stack and montage with EVOS M7000. A549 cells were grown into a spheroid using Nunclon Sphera U-bottom 96-well plates and labeled using SYTOX Deep Red nuclear dye. Z slices were taken at 2 micron steps using an EVOS M7000 Imaging System equipped with an Olympus 10x Super Apochromat objective and a Cy5 EVOS light cube to visualize spheroid morphology in 3D. Images display a montage of individual slices (right) and a mean intensity projection (MPI) of all 190 slices (left).
3D deconvolution, 3D rendering, iso-surface generation and creation of the animation. HepG2 spheroids were grown in a bioreactor and plated on a Nunc glass bottom 96-well plate. Following fixation and permeabilized, spheroids were stained with Hoechst and Alexa Fluor 555 Phalloidin. An EVOS M7000 microscope with 10x S-Apo objective and DAPI and RFP light cubes was used to capture 110 Z-stacks with 2 µm step size. Celleste image analysis software was used for 3D deconvolution, 3D rendering, iso-surface generation and creation of the animation.
EVOS M7000 microscope Z-stack of rat cortical neurons using a 20X S-Apo objective, DAPI, GFP and RFP LED cubes. Rat cortical neurons (Cat. No. A1084001) transfected with CellLight Golgi-RFP, BacMam 2.0 (C10593). At 48 hours cells were fixed and stained with anti-Tau (Cat. No. MN1000), followed by Goat anti-Mouse Alexa Fluor Plus 488 (Cat. No. A32723) and NucBlue Fixed Cell ReadyProbes Reagent (Cat. No. R37606). Cells were imaged on an EVOS M7000 (Cat. No. AMF7000) using an Olympus 20x objective (Cat. No. AMEP4734) with DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651) and RFP (Cat. No. AMEP4652) LED cubes.
EVOS M7000 – 3D deconvolution. A Z-stack of a 16 µm mouse kidney section was acquired on the EVOS M7000 microscope using a 20x S-Apo objective, and DAPI, GFP and TX Red LED cubes. Deconvolution and maxi intensity projection were done using Celleste 5.0 image analysis software with the 3D deconvolution module.
System and hardware specifications
| Optics: | Infinity‐corrected optical system; RMS‐threaded objectives with 45 mm parfocal distance |
| Imaging mode: | Fluorescence, brightfield, color brightfield, and phase contrast |
| Illumination: | Five position chamber for 4 fluorescence light cubes plus brightfield. Light cubes with integrated hard coated filter set and LED light source with >50,000 hour life. Broad selection of standard and specialty light cubes. |
| Imaging methods: | Single color, multi-color, area scan with montage or tile-stitch, time lapse, Z-stacking, movie capture |
| Objective capacity: | 5‐position turret |
| Objectives (not included): | Wide selection of high‐quality LWD and coverslip‐corrected objectives |
| Condenser: | 60 mm long working distance condenser, 4 position turret with a clear aperture and 3 phase annuli. |
| Stage: | Motorized X/Y scanning stage. Travel range 120 mm x 80 mm with sub‐micron resolution, drop‐in inserts to receive vessel holders and lock down holders to fix sample in place during long scans. |
| Focus Mechanism: | Automated focus mechanism with sub‐micron resolution |
| LCD display: | 23" high‐resolution touch screen color monitor (also fully controllable via mouse); 1920x1080 pixel resolution. |
| Cameras: |
|
| Computer: | External Dell PC with an Intel Core i7-8700 processor, 32 GB DDR4 RAM, 512 GB PCIe SSD and NVIDIA Quadro P1000 with 4GB discrete video graphics running Windows® 10, designed to operate with touchscreen monitor and microscope |
| Captured Images: | 16‐bit RAW monochrome: TIFF, PNG 8-bit TIFF, PNG, JPG Movies and time‐lapse: AVI, WMV |
| Output ports: | Computer: 1 x USB 3.1 Gen 2 Type-C; 5 x USB 3.1 Gen 1 Type-A; 4 x USB 2.0 Type-A; 1 Serial; 2 Display Ports 1.2; 1 RJ-45; 2 PS/2; 1 UAJ; 1 Line-out |
| Networking capability: | Connection through Windows/SMB network via an Ethernet cable connection |
| Power supply: | 24 V AC adapter with country‐specific power cords |
| Dimensions (LxWxH) | 18x14x13 inches 457x330x356 mm |
| Weight: | 16/35 (kg/lb) |
Powerful, intuitive software
All EVOS microscopes come with onboard software that allows you to view, capture, and process images quickly and easily. Featuring a user interface and workflow designed by biologists for biologists, the software gives you full control over your fluorescence, brightfield, color brightfield, and phase contract images.
On the M7000 and M5000, the onboard software offers pinpoint operational control and powerful image processing tools such as cell counting, confluence, Z-stack visualization, and time-lapse movies. These tools operate intuitively with minimal training, aided by onboard SmartStart guides. Autofocus and other automated tools save time and effort. The M7000 adds image tiling and stitching as well as fully automated capabilities for image acquisition and processing when scanning tissue slices or acquiring data from multiwell plate vessels. The ability to analyze images in batch mode allows you to seamlessly process large quantities of data.
Click image to enlarge
Fast, robust autofocus
The autofocus algorithm on the EVOS M7000 and M5000 is fast and efficient in any channel. For automated routines with the M7000, you can choose the interval at which autofocus is used in an area scan or multiwell plate acquisition.
Click image to enlarge
Cell confluence app
Automatically measure and document the percent confluence of your cultured cells, allowing consistent passaging and increasing reproducibility of downstream cell-based assays. Use it for a single field or in batch mode for multiple fields and wells.
Click image to enlarge
Automatic cell counting
Advanced tools let you identify and count fluorescent objects while gating out undesired objects based on size, brightness, and circularity. Apply to a single field or to a multiwell plate using batch analysis.
Click image to enlarge
Measure transfection efficiency in cultured cells
Automatically calculate transfection efficiency based on a user-adjustable threshold of positive pixels in a transmitted light or fluorescence image. Apply to a single field or to multiple fields using batch analysis.
Images acquired with EVOS M7000 and deconvolution performed using Celleste 5.0
Click image to enlarge
Figure 1. U2OS cells labeled with Alexa Fluor Plus 555 Phalloidin and imaged with EVOS M7000. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), Tubulin (Cat. No. 32-2600) detected using Goat anti mouse Alexa Fluor Plus 647 secondary (Cat. No. A32728), and Alexa Fluor Plus 555 Phalloidin (Cat. No. A30106) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 2. HeLa cells labeled with Alexa Fluor Plus 555 Phalloidin and imaged with EVOS M7000. HeLa cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605) and Alexa Fluor Plus 555 Phalloidin (Cat. No. A30106) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650) and RFP (Cat. No. AMEP4652) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 3. HeLa cells labeled with Alexa Fluor Plus 555 Phalloidin and imaged with EVOS M7000. HeLa cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605) and Alexa Fluor Plus 555 Phalloidin (Cat. No. A30106) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650) and RFP (Cat. No. AMEP4652) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 4. HeLa cells labeled with Alexa Fluor Plus 647 Phalloidin and imaged with EVOS M7000. HeLa cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605) and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60 Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650) and Cy5 (Cat. No. AMEP4656) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 5. HeLa cells labeled with Alexa Fluor Plus 647 Phalloidin and imaged with EVOS M7000. HeLa cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605) and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60 Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650) and Cy5 (Cat. No. AMEP4656) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Click image to enlarge
Figure 6. U2OS cells labeled with Alexa Fluor Plus 647 Phalloidin and imaged with EVOS M7000. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), CellLight Mitochondria-RFP BacMam 2.0 (Cat. No. C10601), and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes. Deconvolution was performed using Celleste 5.0 Software.
Images acquired with EVOS M7000 and Tissue Stitching
Click image to enlarge
Figure 7. Sagittal Rat brain section detection of neuronal and glial markers and Tissue Stitching in SlowFade Glass. 10 mm cryo-tissue of sagittal section of mouse brain was fixed in PBS buffered 4% paraformaldehyde for 10 min. Afterward the tissue was permeabilized and cleaned in 1% Triton/PBS for 30 minutes. The section was then blocked with 10% normal goat serum followed by additional blocking with ReadyProbes mouse on mouse IgG blocking solution (Cat. No. R37621). Primary antibody of mouse monoclonal against HuC/HuD (Cat. No. A21271) was incubated overnight at 4˚C. Subsequentially, the tissue was labeled using the Alexa Fluor 647, goat anti-mouse secondary HuC/D detection (1000x dilution for 30 min). Tissue was mounted with SlowFade Glass Soft-set Antifade Mountant, with DAPI (Cat. No. S36920), imaging (4x Olympus objective) and tissue stitching was performed on EVOS M7000 Imaging System.
Click image to enlarge
Figure 8. A stitched image showing the live cell density in an entire well of a 6-well plate whole well stitching. Live HeLa cells were stained with calcein AM and imaged with EVOS M7000 using 10x objective. 13x18 stitched image (234 FOV) shows an entire well of a 6-well plate. Note the resolution of the individual FOV is retained when stitching an entire well.
Click image to enlarge
Figure 9. A stitched image showing the fixed cell density in an entire well of a 6-well plate whole well stitching. HeLa cells were fixed with 4% PFA, stained with Eosin-Y and imaged with EVOS M7000 using 10x objective. 13x18 stitched image (234 FOV) shows an entire well of a 6-well plate. Note the resolution of the individual FOV is retained when stitching an entire well.
Click image to enlarge
Figure 10. 384-well spheroid screening. A549 spheroids imaged label-free (brightfield) on an EVOS M7000 using a 10x Olympus SApo objective in a 384-well plate to demonstrate the speed and efficacy of 384-well screening.
Images acquired with EVOS M7000
Click image to enlarge
Figure 11. U2OS cells labeled with Alexa Fluor Plus 555 Phalloidin and imaged with EVOS M7000. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), Tubulin (Cat. No. 32-2600) detected using Goat anti mouse Alexa Fluor Plus 647 secondary (Cat. No. A32728), and Alexa Fluor Plus 555 Phalloidin (Cat. No. A30106) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes.
Click image to enlarge
Figure 12. U2OS cells labeled with Alexa Fluor Plus 647 Phalloidin and imaged with EVOS M7000. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), CellLight Mitochondria-RFP BacMam 2.0 (Cat. No. C10601), and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes.
Olympus 10x Apo objective; transmitted light only; 3x3 tile per well; 3,456 images in 21 mins using EVOS M7000.
DMSO (untreated)
1 µM Jasplakinolide-treated
Brightfield stitch time lapse. U2OS cells monitored label-free over 72 hours, imaged every 30 mins on an EVOS M7000 with an Olympus 10x S-Apo objective in a 37C 5% CO2 incubated environment using an EVOS On-Stage Incubator. Image shown is a 2x2 stitch of DMSO (negative control) treated versus cells treated with 1 µM of the actin-arresting drug Jasplakinolide. While normal cell proliferation can be seen in the control condition, membrane blebbing and internal vesicle transport arrest can be seen in the treated cells due to the arrest of the actin cytoskeleton;
Single FOV
Stitch time lapse
Time-lapse video stitching. Human monocyte-derived macrophages were labeled in a Mattek dish and labeled Ramos added to macrophages along with Hoecsht 33342 and 10nM Rituximab. Red signal indicates phagocytosis and acidification of Ramos cells by macrophages. Images acquired on EVOS M7000 (Cat. No. AMF7000) using a 20x EVOS Fluorite Phase (Cat. No. AMEP4982) air objective. Image shown is a stitched composite of 25 fields of view.
U2OS cells labeled with Alexa Fluor Plus 647 Phalloidin, imaged with EVOS M7000 and deconvolution performed with Celleste 5.0 software. U2OS cells labeled using NucBlue Live ReadyProbes Reagent (Cat. No. R37605), CellLight Talin-GFP BacMam 2.0 (Cat. No. C10611), CellLight Mitochondria-RFP BacMam 2.0 (Cat. No. C10601), and Alexa Fluor Plus 647 Phalloidin (Cat. No. A30107) show superb multiplexing capability and staining specificity. Cells were mounted in ProLong Glass Antifade Mountant (Cat. No. P36984). Images were generated using an EVOS M7000 Imaging System (Cat. No. AMF7000) with an Olympus 60x Aprochromat Oil objective (Cat. No. AMEP4694) using DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651), RFP (Cat. No. AMEP4652), and Cy5 (Cat. No. AMEP4656) EVOS light cubes.
Z-stack animation
Z-stack montage
Click image to enlarge
Z-stack animation Z- stack and montage with EVOS M7000. A549 cells were grown into a spheroid using Nunclon Sphera U-bottom 96-well plates and labeled using SYTOX Deep Red nuclear dye. Z slices were taken at 2 micron steps using an EVOS M7000 Imaging System equipped with an Olympus 10x Super Apochromat objective and a Cy5 EVOS light cube to visualize spheroid morphology in 3D. Images display a montage of individual slices (right) and a mean intensity projection (MPI) of all 190 slices (left).
3D deconvolution, 3D rendering, iso-surface generation and creation of the animation. HepG2 spheroids were grown in a bioreactor and plated on a Nunc glass bottom 96-well plate. Following fixation and permeabilized, spheroids were stained with Hoechst and Alexa Fluor 555 Phalloidin. An EVOS M7000 microscope with 10x S-Apo objective and DAPI and RFP light cubes was used to capture 110 Z-stacks with 2 µm step size. Celleste image analysis software was used for 3D deconvolution, 3D rendering, iso-surface generation and creation of the animation.
EVOS M7000 microscope Z-stack of rat cortical neurons using a 20X S-Apo objective, DAPI, GFP and RFP LED cubes. Rat cortical neurons (Cat. No. A1084001) transfected with CellLight Golgi-RFP, BacMam 2.0 (C10593). At 48 hours cells were fixed and stained with anti-Tau (Cat. No. MN1000), followed by Goat anti-Mouse Alexa Fluor Plus 488 (Cat. No. A32723) and NucBlue Fixed Cell ReadyProbes Reagent (Cat. No. R37606). Cells were imaged on an EVOS M7000 (Cat. No. AMF7000) using an Olympus 20x objective (Cat. No. AMEP4734) with DAPI (Cat. No. AMEP4650), GFP (Cat. No. AMEP4651) and RFP (Cat. No. AMEP4652) LED cubes.
EVOS M7000 – 3D deconvolution. A Z-stack of a 16 µm mouse kidney section was acquired on the EVOS M7000 microscope using a 20x S-Apo objective, and DAPI, GFP and TX Red LED cubes. Deconvolution and maxi intensity projection were done using Celleste 5.0 image analysis software with the 3D deconvolution module.
System and hardware specifications
| Optics: | Infinity‐corrected optical system; RMS‐threaded objectives with 45 mm parfocal distance |
| Imaging mode: | Fluorescence, brightfield, color brightfield, and phase contrast |
| Illumination: | Five position chamber for 4 fluorescence light cubes plus brightfield. Light cubes with integrated hard coated filter set and LED light source with >50,000 hour life. Broad selection of standard and specialty light cubes. |
| Imaging methods: | Single color, multi-color, area scan with montage or tile-stitch, time lapse, Z-stacking, movie capture |
| Objective capacity: | 5‐position turret |
| Objectives (not included): | Wide selection of high‐quality LWD and coverslip‐corrected objectives |
| Condenser: | 60 mm long working distance condenser, 4 position turret with a clear aperture and 3 phase annuli. |
| Stage: | Motorized X/Y scanning stage. Travel range 120 mm x 80 mm with sub‐micron resolution, drop‐in inserts to receive vessel holders and lock down holders to fix sample in place during long scans. |
| Focus Mechanism: | Automated focus mechanism with sub‐micron resolution |
| LCD display: | 23" high‐resolution touch screen color monitor (also fully controllable via mouse); 1920x1080 pixel resolution. |
| Cameras: |
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| Computer: | External Dell PC with an Intel Core i7-8700 processor, 32 GB DDR4 RAM, 512 GB PCIe SSD and NVIDIA Quadro P1000 with 4GB discrete video graphics running Windows® 10, designed to operate with touchscreen monitor and microscope |
| Captured Images: | 16‐bit RAW monochrome: TIFF, PNG 8-bit TIFF, PNG, JPG Movies and time‐lapse: AVI, WMV |
| Output ports: | Computer: 1 x USB 3.1 Gen 2 Type-C; 5 x USB 3.1 Gen 1 Type-A; 4 x USB 2.0 Type-A; 1 Serial; 2 Display Ports 1.2; 1 RJ-45; 2 PS/2; 1 UAJ; 1 Line-out |
| Networking capability: | Connection through Windows/SMB network via an Ethernet cable connection |
| Power supply: | 24 V AC adapter with country‐specific power cords |
| Dimensions (LxWxH) | 18x14x13 inches 457x330x356 mm |
| Weight: | 16/35 (kg/lb) |
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