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Digital image integrity

While a certain degree of image processing is unavoidable for some experiments, fields and techniques, in all instances, the final image presented in the published article must accurately represent the original data and conform to community standards. Authors must take care to exercise caution during data acquisition to avoid misrepresentation of the data collected.

Editors may request the unprocessed data files to help in manuscript evaluation during the peer review process; if these data are unavailable upon request, we may need to halt the peer review process until the issues are satisfactorily resolved. 

We may also request unprocessed data when responding to post-publication issues that may arise with published papers. Lack of availability of unprocessed data can make resolution of post-publication issues challenging. We strongly recommend retaining unprocessed data and metadata files after publication, ideally archiving data in perpetuity. 

Some journals may require submission of unprocessed original images of gels and blots to be submitted with the final accepted version. These unprocessed images are then published in the Supplementary Information.

1. General image guidelines 

  • Do not combine images taken at different times or locations unless clearly indicated as time-averaged or a time-lapse sequence. Clearly separate adjacent images with borders and label and describe each image in the figure legend.
  • Avoid using tools that deliberately alter images (e.g., cloning, healing).
  • Apply adjustments (brightness, contrast, etc.) uniformly across the entire figure panel, including controls. Do not alter data visibility or selectively emphasize regions.
  • Be prepared to provide original, unprocessed or uncropped images upon request. In the event this is not possible, be prepared to present  experimental replicates of the images presented. ​​​​​​

2. Image processing and acquisition tools

  • List all image acquisition tools and software used, including version numbers.
  • If custom code is used for processing, provide a full description and make it available in a community repository.
  • Fully document all image acquisition settings and processing steps in the Methods section.

3. Electrophoretic gels and blots

  • Cropping is acceptable if it improves clarity but mention this in the figure legend. Some journals may require the original, uncropped images in supplementary files.
  • Avoid comparing samples across different gels/blots. If necessary, note that the samples are from the same experiment and processed in parallel.
  • Use clear separations (e.g., black lines) for cropped together non-adjacent lanes. Include loading controls on the same blot where possible.
  • Do not overexpose gels/blots to prevent loss of faint bands. If high contrast is needed, provide multiple exposures in the Supplementary Information.
  • All papers containing blots should provide uncropped images with the membrane edges clearly visible as a supplementary information file ready for peer-review.

4. Microscopy

  • Do not group cells from different fields in one image; use Supplementary Information to show additional fields.
  • Apply image adjustments uniformly. Any use of pseudo-colouring, threshold adjustments, or gamma changes must be disclosed in the figure legend.
  • Ensure all microscopy images are presented with scale bars and/or indicated magnification. Images from different groups shown side-by-side should be presented at the same magnification to allow accurate visual comparison.
  • Specify equipment details (e.g., microscopes, cameras, lenses) and list key acquisition settings in the Methods section. Mention any processing software used and describe manipulations such as deconvolution, 3D reconstructions, or thresholding.
  • State the image resolution and any processing done to enhance it.
  • All colour scales must be defined and intensity levels must be provided in either the figure or its associated legend.

These guidelines help ensure that images accurately represent your data, maintaining transparency and trust in your published research.