Question: What are the imaging system requirements for running Visium for FFPE v1?
Answer: The following imaging system requirements are recommended for running Visium for FFPE v1:
- Microscope features needed:
- Objectives: 10X (NA 0.45), 20X (NA 0.75), or 40X (NA 0.75)
- Motorized tile scanning functionality
- Minimum Capture Resolution 2.18 μm/pixel
- Brightfield Features (for H&E staining):
- Color camera (3 x 8 bit, 2424 x 2424 pixel resolution)
- White balancing functionality
- Exposure times 2 - 10 milliseconds
- Fluorescence Features (for IF staining):
- Light source (or equivalent) with a wavelength range of 380-680 nm
- Monochrome camera (14 bit, 2424 x 2424 pixel resolution)
- DAPI filter cube (Excitation 392/23, Emission 447/60)
- FITC filter cube (Excitation 480/40, Emission 535/50)
- TRITC filter cube (Excitation 542/20, Emission 620/52)
- Cy5 filter cube (Excitation 618/50, Emission 698/70)
- Exposure times 100 milliseconds - 2 seconds
- Additional Specifications:
- 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 (inverted)
- Thermo Fisher Scientific EVOS M7000 (inverted)
- Keyence BZX800 (inverted)
- BioTek Cytation 7 (upright or inverted)
- Leica DMi8 (inverted)
- Leica Aperio Versa 8 (upright)
- MetaSystems Metafer (upright)
- Any equivalent system with the features and specifications listed above may be used.
Please note: if you're using an inverted microscope and you cannot generate quality images while imaging through the slide, you may require an objective with a correction collar. Please contact your imaging system supplier for hardware advice.
Please refer to our Visium for FFPE Imaging Guidelines Technical Note for additional details.
Products: Visium for FFPE