Digital Image Integrity and Standards

High-resolution images are not required at initial submission. When a paper is accepted, the publishing team will request high-resolution files suitable for publication.

All digitized images submitted with the final revision of the manuscript should be 300 DPI if possible.

A certain degree of image processing is acceptable for publication (and for some experiments, fields and techniques is unavoidable), but the final image must correctly represent the original data and conform to community standards. The guidelines below will aid in accurate data presentation at the image processing level; authors must also take care to exercise prudence during data acquisition, where misrepresentation must equally be avoided. Manuscripts should include an 'equipment and settings' section with their methods that describes for each figure the pertinent instrument settings, acquisition conditions and processing changes, as described in this guide.

Authors should list all image acquisition tools and image processing software packages used. Authors should document key image-gathering settings and processing manipulations in the methods.
Authors should list all image acquisition tools and image processing software packages used. Authors should document key image-gathering settings and processing manipulations in the methods.
The use of touch-up tools, such as cloning and healing tools in Photoshop, or any feature that deliberately obscures manipulations, is to be avoided.
Processing (such as changing brightness and contrast) is appropriate only when it is applied equally across the entire image and is applied equally to controls. Contrast should not be adjusted so that data disappear. Excessive manipulations, such as processing to emphasize one region in the image at the expense of others (e.g. through the use of a biased choice of threshold settings), is inappropriate, as is emphasizing experimental data relative to the control.

When submitting revised final figures upon conditional acceptance, authors may be asked to submit original, unprocessed images.

Electrophoretic gels and blots

Positive and negative controls, as well as molecular size markers, should be included on each gel and blot – either in the main figure or an expanded data supplementary figure. For previously characterized antibodies, a citation must be provided. For antibodies less well characterized in the system under study, a detailed characterization that demonstrates not only the specificity of the antibody, but also the range of reactivity of the reagent in the assay, should be published as Supplementary Information.

The display of cropped gels and blots in the main paper is encouraged if it improves the clarity and conciseness of the presentation. In such cases, the cropping must be mentioned in the figure legend and the supplementary information should include full-length gels and blots wherever possible. These uncropped images should be labeled as in the main text and placed in a single supplementary figure. The manuscript's figure legends should state that 'full-length blots/gels are presented in Supplementary Figure X.'

Quantitative comparisons between samples on different gels/blots are discouraged; if this is unavoidable, the figure legend must state that the samples derive from the same experiment and that gels/blots were processed in parallel. Vertically sliced images that juxtapose lanes that were non-adjacent in the gel must have a clear separation or a black line delineating the boundary between the gels. Loading controls must be run on the same blot.
Quantitative comparisons between samples on different gels/blots are discouraged; if this is unavoidable, the figure legend must state that the samples derive from the same experiment and that gels/blots were processed in parallel. Vertically sliced images that juxtapose lanes that were non-adjacent in the gel must have a clear separation or a black line delineating the boundary between the gels. Loading controls must be run on the same blot.
Cropped blots in the body of the paper should retain at least six band widths above and below the band.
Cropped blots in the body of the paper should retain at least six band widths above and below the band.
For quantitative comparisons, appropriate reagents, controls and imaging methods with linear signal ranges should be used.

Microscopy

Authors should be prepared to supply Emres Publishers with original data on request, at the resolution collected, from which their images were generated. Cells from multiple fields should not be juxtaposed in a single field; instead multiple supporting fields of cells should be shown as Supplementary Information.

Adjustments should be applied to the entire image. Threshold manipulation, expansion or contraction of signal ranges and the altering of high signals should be avoided. If 'pseudo-colouring' and nonlinear adjustment (e.g. 'gamma changes') are used, this must be disclosed. Adjustments of individual colour channels are sometimes necessary on 'merged' images, but this should be noted in the figure legend.

We encourage inclusion of the following with the final revised version of the manuscript for publication:

In the methods, specify the type of equipment (microscopes/objective lenses, cameras, detectors, filter model and batch number) and acquisition software used. Although we appreciate that there is some variation between instruments, equipment settings for critical measurements should also be listed.
In the methods, specify the type of equipment (microscopes/objective lenses, cameras, detectors, filter model and batch number) and acquisition software used. Although we appreciate that there is some variation between instruments, equipment settings for critical measurements should also be listed.
The display lookup table (LUT) and the quantitative map between the LUT and the bitmap should be provided, especially when rainbow pseudocolor is used. If the LUT is linear and covers the full range of the data, that should be stated.
Processing software should be named, and manipulations indicated (such as type of deconvolution, three-dimensional reconstructions, surface and volume rendering, 'gamma changes', filtering, thresholding and projection).
Authors should state the measured resolution at which an image was acquired and any downstream processing or averaging that enhances the resolution of the image.