A new approach to dual-color two-photon microscopy with fluorescent proteins.

A new approach to dual-color two-photon microscopy with fluorescent proteins.
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DOI:
10.1186/1472-6750-10-6
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发表时间:
2010-02-02
期刊:
影响因子:
3.5
通讯作者:
Drobizhev M
Drobizhev M
中科院分区:
工程技术3区
文献类型:
--
作者:
Tillo SE;Hughes TE;Makarov NS;Rebane A;Drobizhev M

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对荧光蛋白 (FP) 标记的组织和细胞进行双光子双色成像具有挑战性,因为大多数双光子显微镜一次仅提供一种激光激发波长。目前,由于所使用的FP之间的斯托克斯位移差异较大且双光子吸收(2PA)效率较低,双光子双色成像的方法受到限制。在这里,我们提出了一种双色双光子显微镜的新方法,该方法使用蓝色荧光蛋白的最低能量电子跃迁和红色荧光蛋白的较高能量电子跃迁的同时激发。我们的方法不需要较大的斯托克斯位移差异,并且可以扩展到具有更大 2PA 效率和更优化成像特性的各种 FP 对。
Two-photon dual-color imaging of tissues and cells labeled with fluorescent proteins (FPs) is challenging because most two-photon microscopes only provide one laser excitation wavelength at a time. At present, methods for two-photon dual-color imaging are limited due to the requirement of large differences in Stokes shifts between the FPs used and their low two-photon absorption (2PA) efficiency. Here we present a new method of dual-color two-photon microscopy that uses the simultaneous excitation of the lowest-energy electronic transition of a blue fluorescent protein and a higher-energy electronic transition of a red fluorescent protein. Our method does not require large differences in Stokes shifts and can be extended to a variety of FP pairs with larger 2PA efficiency and more optimal imaging properties.
DOI: 10.1021/jp075545v
发表时间: 2007-12-20
影响因子: 3.3
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