US Patent 7626694 Optical microscopy with phototransformable optical labels

First activation radiation is provided to a sample that includes fluorescent proteins (“FPs”) to activate a first subset of the FPs in the sample. First excitation radiation is provided to the first subset of FPs in the sample to excite at least some of the activated FPs, and radiation emitted from activated and excited FPs within the first subset of FPs is detecting with imaging optics. The first activation radiation is controlled such that the mean volume per activated FPs in the first subset is greater than or approximately equal to a diffraction-limited resolution volume (“DLRV”) of the imaging optics.