2-PHOTON LASER SCANNING FLUORESCENCE MICROSCOPY
2-PHOTON LASER SCANNING FLUORESCENCE MICROSCOPY
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DOI：
10.1126/science.2321027
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发表时间：
1990-04-06
期刊：
影响因子：
56.9
通讯作者：
WEBB, WW
中科院分区：
文献类型：
--
作者：
DENK, W;STRICKLER, JH;WEBB, WW
在激光扫描荧光显微镜中，同时吸收两个光子的分子激发提供了固有的三维分辨率。用一束强聚焦的亚皮秒红色激光脉冲激发具有单光子吸收的紫外荧光团，使活细胞和其他微观物体的荧光图像成为可能。随着激发强度的增加，荧光发射呈二次增长，使得荧光和光漂白被限制在焦平面附近，这与双光子协同激发所期望的一致。该技术还为三维空间分辨光化学，特别是笼状效应分子的光解释放提供了前所未有的能力。
Molecular excitation by the simultaneous absorption of two photons provides intrinsic three-dimensional resolution in laser scanning fluorescence microscopy. The excitation of fluorophores having single-photon absorption in the ultraviolet with a stream of strongly focused subpicosecond pulses of red laser light has made possible fluorescence images of living cells and other microscopic objects. The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation. This technique also provides unprecedented capabilities for three-dimensional, spatially resolved photochemistry, particularly photolytic release of caged effector molecules.