Optical fluorescence microscopy in three dimensions: microtomoscopy

摘要: SUMMARY A unique property of confocal fluorescence microscopy is utilized to obtain scanned images a number samples in three dimensions. This kind for the first time enables direct study 3-D arrangement fluorescently labelled materials living cells. The technique based on home build, confocal, scanning laser microscope. entire volume object mechanically through well-defined focus illuminated, high numerical aperture objective. An identical optical path, arranged confocally, used detect emission from focal volume. A pinhole image plane detection channel prevents light, generated outside volume, reach detector. Consequently true resolution obtained along axis (identified as z-axis) and three-dimensional imaging without need mathematical operations possible. z-axis (0.9 μm) shown well (0.25 using Rhodamine fluorescent dye. explained experimentally demonstrated an which contains only dependent structure. Pictures sections cells are shown, both parallel axis.