Minimum Information guidelines for fluorescence microscopy: increasing the value, quality, and fidelity of image data.

High-resolution digital microscopy provides ever more powerful tools for probing the real-time dynamics of subcellular structures, and adequate record-keeping is necessary to evaluate results, share data, and allow experiments to be repeated. In addition to advances in microscopic techniques, post-acquisition procedures such as image-data processing and analysis (i.e., feature counting, distance measurements, intensity comparison, and colocalization studies) are often required for the reproducible and quantitative interpretation of images. While these techniques increase the usefulness of microscopy data, the limits to which quantitative results may be interpreted are often poorly quantified and documented. Keeping notes on microscopy experiments and calibration procedures should be relatively unchallenging, as the microscope is a machine whose performance should be easy to assess. Nevertheless, to this date, no widely adopted data provenance and quality control metadata guidelines to be recorded or published with imaging data exist. Metadata automatically recorded by microscopes from different companies vary widely and pose a substantial challenge for microscope users to create a good faith record of their work. Similarly, the complexity and aim of experiments using microscopes vary, leading to different reporting and quality control requirements from the simple description of a sample to the need to document the complexities of sub-diffraction resolution imaging in living cells and beyond. To solve this problem, the 4DN Imaging Standards Working Group has put forth a tiered system of microscopy calibration and metadata standards for images obtained through fluorescence microscopy. The proposal is an extension of the OME data model and aims at increasing data fidelity, ease future analysis, and facilitate objective comparison of different datasets, experimental setups, and essays.