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:
- Objectives: 4X (Plan APO λ; NA 0.20); 10X (Plan APO λ; NA 0.45); 20X (Plan APO λ; NA 0.75)
- Motorized tile scanning functionality
- Minimum Capture Resolution 2.18 μm/pixel
Brightfield Features (for H&E staining only):
- Color camera (3 x 8 bit, 2424 x 2424 pixel resolution)
- White balancing functionality
- Exposure times 2 - 10 milliseconds
Fluorescence Features (required for the Tissue Optimization workflow and IF staining):
- Light source (or equivalent) with a wavelength range of 380-680 nm
- Monochrome camera (14 bit, 2424 x 2424 pixel resolution)
- TRITC filter cube (Excitation 542/20, Emission 620/52)
- DAPI filter cube (Excitation 392/23, Emission 447/60) - required for IF staining only
- Exposure times 100 milliseconds - 2 seconds
- Computer with sufficient power to handle large images (0.5-5 GB)
- Image stitching software (microscope’s native software or third-party software, such as Image J)
- Images have a minimum size requirement of 2000 pixels in at least one direction
- Images saved in tiff (preferred) or jpeg format
Imaging systems compatible with Visium:
- Nikon Eclipse Ti2-e
- Thermo Fisher Scientific EVOS M7000
- Hamamatsu NanoZoomer S60
- Keyence BZX800
- BioTek Cytation 7
- Leica DMi8
- Leica Versa 8
- Molecular Devices ImageXpress Nano Automated Cell Imaging System
- 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