OPTICAL MICROSCOPY WITH PHOTOTRANSFORMABLE OPTICAL LABELS

摘要

The invention relates to a method comprising steps of: providing spatially-structured activation radiation having relatively high- and relatively low-intensity regions to a sample that includes phototransformable optical labels ("PTOLs") to activate a subset of the PTOLs in the sample located predominately at relatively high intensity regions of the spatially-structured activation radiation; providing spatially-structured excitation radiation to the subset of activated PTOLs in the sample, wherein the exciting radiation is structured so that one or more relatively high intensity regions of the excitation radiation at least partially overlap one or more relatively high intensity regions of the activating radiation; detecting radiation emitted from the activated and excited PTOLs with imaging optics; and controlling the intensities and spatial structures of the activating radiation and the exciting radiation so that radiation emitted from PTOLs in the sample is emitted substantially from at least one volume that is comparable to or less than a diffraction-limited resolution volume ("DLRV") of the imaging optics. The PTOLs may be formed by photoactivated or photoswitched fluorescent proteins. The invention also relates to a corresponding system.