Scan Multiwell Plates

Imaging Multiwell Plates

Multiwell plates in 6, 12, 24, 48, 96, and 384 well formats can be scanned on the Odyssey M. Be sure to select multiwell plates that fit the Plate Alignment Guide.

Do not attempt to image plates that have a height greater than 23 mm on the Odyssey M.

Proper selection of multiwell plates significantly affects the results of your analysis, because each plate has its own characteristics including well depth, autofluorescence, and well-to-well signal crossover. Some general considerations for multiwell plate selection are provided here.

  • Do not use plates with white walls, because the autofluorescence from the white surface will create significant noise.
  • For an In‑Cell Western™ Assay, LI‑COR recommends the following plates.
    • For adherent cells, LI‑COR recommends a 96-well plate with a clear, flat bottom and black wells, such as the Greiner Bio-One CELLSTAR® Black μClear® Microplate, LI‑COR PN 926-19156 (8 pack) or 926-19157 (32 pack).
    • For suspension cells, LI‑COR recommends growing cells in a 96-well U-bottom plate and transferring cells to Greiner Bio-One CELLSTAR® Black μClear® Microplate, LI‑COR PN 926-19156 (8 pack) or 926-19157 (32 pack), for imaging.
  • The Odyssey M Scan Surface should be thoroughly cleaned using the procedures described earlier in this chapter. If artifacts are noticed on the image, it may help to clean the bottom of the plate with a moist, lint-free tissue.
  • Protect plates from light before imaging to ensure highest sensitivity. When storing plates after imaging, protect plates from light.

  • Plates other than those recommended above may require lower or higher focus offsets for optimal resolution and detection. If alternative plates are used, an initial optimization scan will be necessary. Scan a plate containing experimental and control samples at a range of focus offsets. The focus offset range for the Odyssey M is -1.00 mm to 5.00 mm. Use the same intensity settings for each scan. After reviewing the collected scans, use the focus offset with the highest signal-to-noise ratio as the focus offset for experiments with the alternate plates.

Scanning Multiwell Plates

  1. Ensure the Odyssey M is powered on and connected to LI‑COR® Acquisition Software LI-COR Acquisition icon.

  2. Open the lid.

  3. For the recommended Greiner plates, we generally recommend taking the lid off before imaging. You can use plate tape to cover the plate to mitigate evaporation problems.

    When scanning live cells, you may need to leave the lid on to prevent contamination.

  4. Place the Plate Alignment Guide on the Odyssey M Scan Surface and place multiwell plates in the Plate Alignment Guide.

  5. Close the lid.

  6. In LI‑COR Acquisition Software, choose the workflow for scanning multiwell plates and follow the guidance on the pages in the workflow.