Scan a Membrane

See the links on the right for information about acquiring membrane images on the Odyssey DLx and Odyssey CLx, Odyssey XF and Odyssey Fc, or Odyssey M. For information about chemiluminescence imaging, see Scan a Chemiluminescent Membrane.

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

Scan a Membrane on the Odyssey M Imager

Once connected, click Membrane on the Choose Assay page to begin imaging membranes.

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

  • Resolution: 100 µm

  • Focus Offset: 0.00 mm

Set Scan Area

  • Draw multiple Scan Areas to image multiple membranes.
  • 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

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

Click Scan to begin image acquisition.

  1. Only one non-RGB Epi channel may be selected.

  2. Select up to 4 fluorescent channels.

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, RGB Epi
700 IRDye 680LT, IRDye 680RD, Revert 700 Total Protein Stain, Alexa Fluor 680 685 nm 721 - 740 nm 800, 520, 488, RGB Epi
520 Revert 520 Total Protein Stain, VRDye 549, Cy3, Alexa Fluor 546 520 nm 570 - 610 nm 800, 700, 488, RGB Epi
488 VRDye 490, FITC, Alexa Fluor 488 488 nm 519 - 543 nm 800, 700, 520, RGB Epi
RGB Epi Colorimetric Protein MW Markers NA NA 800, 700, 520, 488, Chemi, or 630 Epi, 525 Epi
630 Epi Pierce Reversible Protein Stain, TMB (blotting) 631 - 659 nm Clear Window RGB Epi
525 Epi Ponceau S, NBT/BCIP 500 - 530 nm Clear Window RGB Epi

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.

Scan a Membrane on the Odyssey® F Imager

Once connected, click Membrane on the Choose Assay page to begin imaging membranes.

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

  • Resolution: 169 µm

  • Focus Offset: 0.00

Set Scan Area

  • Draw multiple Scan Areas to image multiple membranes.
  • 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 Empiria Studio Autofill Button icon 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. 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.

Channels* Labels Excitation Emission
800 IRDye® 800CW, DyLight 800 785 nm 813 – 832 nm
700 IRDye 680LT, IRDye 680RD, Revert 700 Total Protein Stain, Alexa Fluor 680 685 nm 710 – 730 nm
520 Revert™ 520 Total Protein 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 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.

Scan a Membrane on the Odyssey DLx Imager or Odyssey CLx Imager

Once connected, click Membrane on the Choose Assay page to begin imaging membranes.

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

  • Resolution: 169 µm

  • Focus Offset: 0.00 mm

Set Scan Area

  • Draw multiple Scan Areas to image multiple membranes.
  • 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 Empiria Studio Autofill Button icon 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. 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 680LT, IRDye 680RD, Revert 700 Total Protein Stain, Alexa Fluor 680 685 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.

Scan a Membrane on the Odyssey XF Imager or Odyssey Fc Imager

Once connected, click Membrane on the Choose Assay page to begin imaging chemiluminescent membranes.

Image settings for this workflow

  • Resolution: 125 µm

  • Chemi Acquisition Time Options: 30 sec, 1 min, 2 min (default), 5 min, 10 min

  • Chemi Manual Entry for Acquisition Time: 5 seconds to 60 minutes

    Click the button to enter a custom acquisition time from 5 seconds to 60 minutes.

  • Resolution: 125 µm

  • Fluorescence Acquisition Time Options: 30 sec, 1 min, 2 min (default), 5 min, 10 min

  • Manual Entry for Acquisition Time: 5 seconds to 60 minutes

Choose detection options

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 - 846 nm 600, 700, Chemi
700 IRDye 680LT, IRDye 680RD, Revert 700 Total Protein Stain, Alexa Fluor 680 685 nm 716 - 746 nm 600, 800, Chemi
Chemi HRP and AP chemiluminescent substrate NA NA 600, 700, 800
600 Custom 520 nm 573 - 637 nm 700, 800, Chemi

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 Acquire.

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 Repeat Acquire to acquire new image(s) with the same settings.
  • Click Home to disconnect from the imager and return to the Home page.