What are the imaging system requirements for running Visium for fresh-frozen?

Question: What are the imaging system requirements for running Visium for fresh-frozen?

Answer: The imaging system requirements for running Visium are outlined below.

Microscope features needed:

1. Objectives: 4X (Plan APO λ; NA 0.20); 10X (Plan APO λ; NA 0.45); 20X (Plan APO λ; NA 0.75)
2. Motorized tile scanning functionality
3. Minimum Capture Resolution 2.18 μm/pixel

Brightfield Features (for H&E staining only):

1. Color camera (3 x 8 bit, 2424 x 2424 pixel resolution)
2. White balancing functionality
3. Exposure times 2 - 10 milliseconds

Fluorescence Features (required for the Tissue Optimization workflow and IF staining):

1. Light source (or equivalent) with a wavelength range of 380-680 nm
2. Monochrome camera (14 bit, 2424 x 2424 pixel resolution)
3. TRITC filter cube (Excitation 542/20, Emission 620/52)
4. DAPI filter cube (Excitation 392/23, Emission 447/60) - required for IF staining only
5. Exposure times 100 milliseconds - 2 seconds

Additional Specifications:

1. Computer with sufficient power to handle large images (0.5-5 GB)
2. Image stitching software (microscope’s native software or third-party software, such as Image J)
3. Images have a minimum size requirement of 2000 pixels in at least one direction
4. Images saved in tiff (preferred) or jpeg format

Imaging systems compatible with Visium: 

1. Nikon Eclipse Ti2-e
2. Thermo Fisher Scientific EVOS M7000
3. Hamamatsu NanoZoomer S60
4. Keyence BZX800
5. BioTek Cytation 7
6. Leica DMi8
7. Leica Versa 8
8. Molecular Devices ImageXpress Nano Automated Cell Imaging System
9. Any equivalent system with the features and specifications listed above may be used.

For additional details, please refer to our Imaging Guidelines Technical Note.

Product: Visium for fresh-frozen