Scan Multiwell Plates on the Odyssey M Imager

Once connected, click Multiwell Plate on the Choose Assay page to begin imaging a plate-based assay.

Choose Plate Assay

Choose the type of plate assay you want to image.

  • In‑Cell Western - Use this workflow to acquire In‑Cell Western or On-Cell Western plate images.

    Both the In‑Cell Western and On-Cell Western are quantitative immunofluorescence assays performed in microwell plates. Data analysis is based on whole-well fluorescence quantification.

    The following image settings are available for the In-Cell Western option.

    • Resolution for 96-Well plates: 100 µm

    • Resolution for 384-Well plates: 50 µm

    • Focus Offset: 3.8

    • Available Focus Offset Range: 1.00 - 5.00 mm

    Other settings can be entered in the Custom workflow. When using the Custom workflow, keep the following points in mind.

    • Acquisitions with an RGB channel image are not compatible with multiwell plate workflows (In‑Cell Western, Cell Analysis, or ELISA) in Empiria Studio.

    • Epi and trans channel microwell plate images are not compatible with In‑Cell Western or Cell Analysis workflows in Empiria Studio.

  • ELISA - Use this workflow to analyze ELISA (enzyme-linked immunosorbent assay) plate images. Fluorescence microwell plate images are not compatible with analysis in the ELISA workflow in Empiria Studio.

    • Resolution for 96-Well Plates: 100 µm

    • Resolution for 384-Well Plates: 50 µm

    • Focus Offset: 3.00 mm

    • Available Focus Offset Range: 1.00 - 5.00 mm

  • Cell Analysis - Cell Analysis refers to a broad range of assays for studying the function and localization of proteins in living and fixed cells. Cell Analysis covers assays that are not more precisely described as an ELISA or an In-Cell Western Assay (which includes On-Cell Western assays in the context of LI‑COR® Acquisition Software and Empiria Studio® Software).

    The following image settings are available for the Cell Analysis option.

    • Resolution: 100 µm (96-well, 48-well, 24-well, 12-well, 6-well)

    • Focus Offset: 3.00 mm

    • Available Focus Offsets: 1.00 - 5.00 mm

    Other settings can be entered in the Custom workflow. When using the Custom workflow, keep the following points in mind.

    • Acquisitions with an RGB channel image are not compatible with multiwell plate workflows (In‑Cell Western, Cell Analysis, or ELISA) in Empiria Studio.

    • Epi and trans channel microwell plate images are not compatible with In‑Cell Western or Cell Analysis workflows in Empiria Studio.

Select plate locations

Select the location of your plate(s) on the Odyssey M Scan Surface.

Choose detection options

  1. If you have multiple Scan Areas, click Edit Channels to choose detection options independently for each plate.

  2. Select the plate's well format.

  3. Click Scan to begin image acquisition.

Detection label reference

The detection labels listed in this table are examples. If you do not see the detection label you want to use, check its spectral characteristics to make sure it is compatible.

Channel Labels Excitation Emission Combined Selections Allowed
800 IRDye® 800CW, DyLight 800 785 nm 816 - 840 nm 700, 520, 488
700 IRDye 680RD, CellTag™ 700 Stain, Sapphire700™ Stain, DRAQ5 685 nm 721 - 740 nm 800, 520, 488
520 VRDye 549, Cy3, Alexa Fluor 546, CellTag 520 Stain 520 nm 570 - 610 nm 800, 700, 488
488 VRDye 490, FITC, Alexa Fluor 488 488 nm 519 - 543 nm 800, 700, 520
Channel Labels Excitation Emission Combined Selections Allowed
630 Trans TMB - non stopped 631 - 659 nm Clear Window None
470 Trans OPD, TMB - Stopped 447 - 472 nm Clear Window None
Channel Labels Excitation Emission Combined Selections Allowed
800 IRDye® 800CW, DyLight 800 785 nm 816 - 840 nm 700, 520, 488
700 IRDye 680RD, CellTag™ 700 Stain, Sapphire700™ Stain, DRAQ5, SYTO 60 685 nm 721 - 740 nm 800, 520, 488
520 VRDye 549, Cy3, Alexa Fluor 546, Red fluorescent protein (RFP), Propidium Iodide 520 nm 570 - 610 nm 800, 700, 488
488 VRDye 490, Green Fluorescent Protein (GFP), NucSpot 470, SYTOX Green 488 nm 519 - 543 nm 800, 700, 520

Scan

After clicking Scan on the Choose detection options page, the Scan page will open with the image acquisition process already started.

When the scan has completed, the Ready symbol will appear .

Stop Scan

If you need to stop an image acquisition before the acquisition process completes, click Stop Scan.

This will stop all pending Acquisitions and will remove any partially completed Acquisitions.

Export Acquisition

Ensure the Include all acquisitions option is selected to export every image for every channel from this acquisition process.

Export to Analyze

Click Export to Analyze to export images to a single file. Import the file into Empiria Studio® Software for analysis.

Export to Publish or Present

Click Export to Publish or Present to export the current image view to an image file.

Images can be exported as high resolution TIFF files (for journal publications) or as PNG files appropriate for digital presentation (such as a slide show).

The default export image size shown in the "Export" dialog is the largest dimension (i.e., width or height) of the image set to 10 cm (4 in), and the other dimension is set according to the aspect ratio of the image. If you change the width or height in the dialog, the other dimension will update automatically to maintain the correct aspect ratio.

Workflow

  • Click Start New to return to the beginning of the workflow to acquire another multiwell plate image.
  • Click Leave to disconnect from the imager and return to the Getting Started page.