FLUORESCENT NANOSCOPY METHOD (VARIANTS) Russian patent published in 2007 - IPC G01N21/00 G01N21/64 

FIELD: analysis of objects, colored by fluorescent colorants, using a modified fluorescent microscope.

SUBSTANCE: in accordance to the invention, method uses fluorescent microscope, video cameras, a computer. Microscope contains digital cameras with locking filters for singling out the light of fluorescence. Structures of object or its cut are colored with one or different non-fluorescent colorants. When object is lit with an ultraviolet flash, some of the non-fluorescent molecules of colorant are transformed to fluorescent ones. Result is recorded in form of separate spots on video camera frames, which are transferred to the computer. Fluorescent molecules decolorized in the process of or after the registration. Frames with remaining fluorescent radiation are transferred into the computer. On basis of difference frames, spot centers, representing coordinates of fluorescent colorant molecules, are searched for. The process is repeated until coordinates of colorant molecules in lit part of object are computed. Computer reconstruction of image of the whole object or its cut is performed. Objects may be colored with fluorescent nano-particles of one or different types.

EFFECT: production of two-dimensional and three-dimensional image with resolution better than 20 nm, possible registration of colored image with coloring by various colorants of proteins, nucleic acids, lipids, etc.

2 cl, 3 dwg