FLUORESCENT NANOSCOPY METHOD

The invention relates to analysis of object dyed with fluorescent colouring agents carried out with the aid of a fluorescent microscope which is modified in terms of improved resolving power and called a nanoscope thereafter. The inventive method is carried out with the aid of a microscope comprising an optical system for visualising and projecting a sample image to a video camera which records and digitises images of individual fluorescence molecules and nanoparticles at a low noise, a computer for recording and processing images, a sample holder arranged in front of an object lens, a fluorescent radiation exciting source and a set of replaceable suppression filter for separating the sample fluorescent light. The invention is characterised in that the separately fluorescing visible molecules and nanoparticles are periodically formed in different object parts, the laser produces the oscillation thereof which is sufficient for recording the non-overlapping images of said molecules and nanoparticles and for decolouring already recorded fluorescent molecules, wherein tens of thousands of pictures of recorded individual molecule and nanoparticle images (in the form of stains whose diameter is of the order of a fluorescent light wavelength multiplied by a microscope amplification) are processed by a computer for searching the co-ordinates of the stain centres and building the object image according to millions of calculated stain center co-ordinates corresponding to the co-ordinates of the individual fluorescent molecules and nanoparticles. Said invention makes it possible to obtain a two- and three-dimensional image with a resoling power better than 20 nm and to record a colour image by dyeing proteins, nucleic acids and lipids with different colouring agents. Le procédé se rapporte à l'analyse d'objets qui sont peints avec des colorants fluorescents, ladite analyse s'effectuant au moyen d'un microscope à fluorescence amélioré qui, grâce à sa résolution améliorée, est parfois appelé "nanoscope". L'invention proposé un procédé pour obtenir une image au moyen d'un microscope qui contient: un système optique d'observation visuelle et de projection de l'image d'un échantillon dans une caméscope capable d'enregistrer et de numériser avec un faible niveau de bruit les images de molécules et de nanoparticules fluorescentes isolées, un ordinateur destiné à l'enregistrement et au traitement d'images, un support d'échantillons, disposé en face de l'objectif, une source