Acquiring a good image requires having access to a well characterized, well calibrated imaging system. In this matter, procedures exist, aimed at measuring relevant parameters which helps testing, following and comparing microscopes performances. In this paper, we focus on four indicators : the system resolution, the characterization of its chromatic aberration, evaluation of the field illumination homogeneity and finally the estimation of the detector associated noise (CV : coefficient of variation). The built of reference samples is discussed, and the subsequent image acquisition procedures are proposed as charts. We introduce for the first time the MetroloJ plugin, designed to retrieve the four numerical indicators out of images, and to generate archivable reports as pdf files, thanks to the iText library. Altogether, the procedures and the MetroloJ plugin, already tested within the French technological network for multi- dimensional fluorescence microscopies (RT-MFM), provide the user with a mean to keep track of microscopes’ health, and make it a valuable tool in choosing a microscope setup based on pre-defined expectations.
Metrology, microscope calibration, image analysis, fluorescence microscopy
Presenting author: Fabrice P. Cordelieres
Organisation: Institut Curie/CNRS UMR3348 - Plateforme IBiSA d'Imagerie Cellulaire et Tissulaire
co-authors: Cedric Matthews, email@example.com, CNRS-IBDML-UMR 6216, Service Imagerie