Szmacinski H, Gryczynski I, Lakowicz J R
Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore 21201, USA.
Biospectroscopy. 1998;4(5):303-10. doi: 10.1002/(SICI)1520-6343(1998)4:5%3C303::AID-BSPY2%3E3.0.CO;2-X.
We describe spatially localized two-photon excitation in scattering media. Using femtosecond pulses at 770 nm from a Ti: Sapphire laser, we were able to excite fluorophores in capillary tubes under up to 1.5 mm of 0.5% intralipid. Displacement of the laser beam relative to the embedded samples indicates that highly localized excitation was possible with two-photon excitation, whereas one-photon excitation resulted in loss of spatial resolution due to excitation by the diffusely scattered photons. These results indicate that two-photon excitation in the scattering solution is due only to the ballistic photons, a result confirmed by frequency-domain time-resolved measurements. Selective excitation of adjacent embedded samples was found possible for two but not one-photon excitation.
我们描述了散射介质中的空间局域双光子激发。使用钛宝石激光器产生的770纳米飞秒脉冲,我们能够在高达1.5毫米厚的0.5%脂质乳剂下激发毛细管中的荧光团。激光束相对于嵌入样品的位移表明,双光子激发能够实现高度局域的激发,而单光子激发由于漫散射光子的激发而导致空间分辨率丧失。这些结果表明,散射溶液中的双光子激发仅归因于弹道光子,频域时间分辨测量证实了这一结果。结果发现,对于双光子激发而非单光子激发,可以选择性地激发相邻的嵌入样品。