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RESOLFT, an acronym for REversible Saturable OpticaL Fluorescence Transitions, denotes a group of optical microscopy techniques with very high resolution. Using standard far field visible light optics a resolution far below the diffraction limit down to molecular scales can be obtained. With conventional microscopy techniques, it is not possible to distinguish features that are located at distances less than about half the wavelength used (i.e. about 200 nm for visible light). This diffraction limit is based on the wave nature of light. In conventional microscopes the limit is determined by the used wavelength and the numerical aperture of the optical system. The RESOLFT concept surmounts this limit by temporarily switching the molecules to a state in which they cannot send a (fluorescence-) signal upon illumination. This concept is different from for example electron microscopy where instead the used wavelength is much smaller. ==Working principle== RESOLFT microscopy is an optical microscopy with very high resolution that can image details in samples that cannot be imaged with conventional or confocal microscopy. Within RESOLFT the principle of STED microscopy and GSD microscopy are generalized. Structures that are normally too close to each other to be distinguished are read out sequentially. Within this framework all methods can be explained which operate on molecules that have at least two distinguishable states, where reservible switching between the two states is possible, and where at least one such transition can be optically induced. In most cases fluorescent markers are used, where one state (A) which is bright, that is, generates a fluoresence signal, and the other state (B) is dark, and gives no signal. One transition between them can be induced by light (e.g. A→B, bright to dark). The sample is illuminated inhomogeneously with the illumination intensity at one position being very small (zero under ideal conditions). Only at this place are the molecules never in the dark state B (if A is the pre-existing state) and remain fully in the bright state A. The area where molecules are mostly in the bright state can be made very small (smaller than the conventional diffraction limit) by increasing the transition light intensity (see below). Any signal detected is thus known to come only from molecules in the small area around the illumination intensity minimum. A high resolution image can be constructed by scanning the sample, i.e., shifting the illumination profile across the surface.〔See also Confocal laser scanning microscopy.〕 The transition back from B to A can be either spontaneous or driven by light of another wavelength. The molecules have to be switchable several times in order to be present in state A or B at different times during scanning the sample. The method also works if the bright and the dark state are reversed, one then obtains a negative image. 抄文引用元・出典: フリー百科事典『 ウィキペディア（Wikipedia）』 ■ウィキペディアで「RESOLFT」の詳細全文を読む スポンサード リンク
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