Scan a Plate

See the links on the right for information about acquiring multiwell plate images on the Odyssey DLx and Odyssey DLx or Odyssey M.

See the links below for emission/excitation data and example detection labels.

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

  • Absorbance Assay

    • 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 one of the other Multiwell Plate Assay types.

    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

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
800 IRDye® 800CW, DyLight 800 785 nm 816 - 840 nm
700 IRDye 680RD, CellTag™ 700 Stain, Sapphire700™ Stain, DRAQ5 685 nm 721 - 740 nm
520 VRDye 549, Cy3, Alexa Fluor 546, CellTag 520 Stain 520 nm 570 - 610 nm
488 VRDye 490, FITC, Alexa Fluor 488 488 nm 519 - 543 nm
Chemi HRP and AP substrates, luminescent assays NA NA

Any combination of up to four of the channels in the table can be selected per plate per acquisition. Different channels can be selected for different plates.

Channel Labels Excitation Emission
630 Trans TMB - non stopped, Bradford Protein Assay, Pierce 660nm Protein Assay, Modified Lowry Protein Assay 631 - 659 nm Clear Window
525 Trans OPD, BCA Protein Assay 500 - 530 nm Clear Window
470 Trans OPD, TMB - stopped, BCA Protein Assay 447 - 472 nm Clear Window
RGB Trans True color Transmissive RGB Color
Channel Labels Excitation Emission
800 IRDye® 800CW, DyLight 800 785 nm 816 - 840 nm
700 IRDye 680RD, CellTag™ 700 Stain, Sapphire700™ Stain, DRAQ5, SYTO 60 685 nm 721 - 740 nm
520 VRDye 549, Cy3, Alexa Fluor 546, Red fluorescent protein (RFP), Propidium Iodide 520 nm 570 - 610 nm
488 VRDye 490, Green Fluorescent Protein (GFP), SYTOX Green 488 nm 519 - 543 nm
Chemi HRP and AP substrates, luminescent assays NA NA

Any combination of up to four of the channels in the table can be selected per plate per acquisition. Different channels can be selected for different plates.

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 image file(s). Images exported using this method are not intended for analysis.

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).

When you are exporting a single image, 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.

See View Images in the Image Gallery for more information.

Workflow
  • Click Start New to return to the beginning of the workflow to acquire another multiwell plate image.
  • Click Rescan to acquire new image(s) with the same settings.
  • Click Home to disconnect from the imager and return to the Home page.

Scan a Plate on the Odyssey® F Imager

Once connected, click Multiwell Plate on the Choose Assay page to begin imaging gels.

The following imaging settings are used in this workflow. Other settings can be entered in the Custom workflow.

  • Resolution options:

    • 96-well plates: 84 µm, 169 µm
    • 384-well plates: 84 µm

  • Focus Offset: 1 – 5 mm

  • Well formats: 96 well, 384 well

  • Resolution options: 84 µm, 169 µm

  • Focus Offset: 1 – 5 mm

  • Well Formats: 6 well, 12 well, 24 well, 48 well, 96 well

Set Scan Area

Use the four buttons that correspond to the four locations on the Odyssey F plate holder to designate which plates you want to scan.

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
800 IRDye® 800CW, DyLight 800 785 nm 813 – 832 nm
700 IRDye 680RD, CellTag™ 700 Stain, Sapphire700™ Stain, DRAQ 5 685 nm 710 – 730 nm
520 CellTag™ 520 Stain, VRDye™ 549 Secondary Antibodies, Cy3, Alexa Fluor 546 520 nm 570 – 610 nm
488 VRDye™ 490 Secondary Antibodies, FITC, Alexa Fluor 488 488 nm 538 – 552 nm

*All channels shown are available in this workflow for the 4-laser Odyssey F (9180). The 700 and 800 channels are available for the 2-laser Odyssey F (9150).

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 image file(s). Images exported using this method are not intended for analysis.

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).

When you are exporting a single image, 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.

See View Images in the Image Gallery for more information.

Workflow
  • Click Start New to return to the beginning of the workflow to acquire another gel image.
  • Click Rescan to acquire new image(s) with the same settings.
  • Click Home to disconnect from the imager and return to the Home page.

Scan a Plate on the Odyssey DLx Imager or Odyssey CLx 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 multiwell plates. Data analysis is based on whole-well fluorescence quantification.

    The following image settings are used for the In-Cell Western option. Other settings can be entered in the Custom workflow.

    • Resolution for 96-Well Plate: 169 µm

    • Resolution for 384-Well Plate: 84 µm

    • Resolution Options: 84, 169 µm (default)

    • Focus Offset: 4.00 mm

    • Available Focus Offset Range: 0.00 – 4.00 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 one of the other Multiwell Plate Assay types.

    The following image settings are used for the Cell Analysis option. Other settings can be entered in the Custom workflow.

    • Resolution: 169 µm

    • Resolution Options: 84, 169 µm (default)

    • Focus Offset: 4.00 mm

    • Available Focus Offset Range: 0.00 – 4.00 mm

Set Scan Area

  • Draw multiple Scan Areas to image multiple plates.
  • Adjust the exact size and location of a Scan Area. Select a Scan Area, then adjust the number in the X location, Y location, Width, and Height fields in the Control Panel.

Choose detection options

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

    Click the Autofill button next to detection options for one Scan Area to apply the options for that Scan Area to all Scan Areas below it in the list.

  2. Select the plate 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 812 – 832 nm 700
700 IRDye 680RD, CellTag™ 700 Stain, Sapphire700™ Stain, DRAQ5 685 nm 710 – 730 nm 800
Channel Labels Excitation Emission Combined Selections Allowed
800 IRDye® 800CW, DyLight 800 785 nm 812 – 832 nm 700
700 IRDye 680RD, CellTag™ 700 Stain, Sapphire700™ Stain, DRAQ5 685 nm 710 – 730 nm 800

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 image file(s). Images exported using this method are not intended for analysis.

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).

When you are exporting a single image, 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.

See View Images in the Image Gallery for more information.

Workflow
  • Click Start New to return to the beginning of the workflow to acquire another membrane image.
  • Click Rescan to acquire new image(s) with the same settings.
  • Click Home to disconnect from the imager and return to the Home page.