Live imaging using adaptive optics with fluorescent protein guide-stars.
Live imaging using adaptive optics with fluorescent protein guide-stars.
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DOI：
10.1364/oe.20.015969
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发表时间：
2012-07-02
期刊：
影响因子：
3.8
通讯作者：
Kubby J
中科院分区：
文献类型：
--
作者：
Tao X;Crest J;Kotadia S;Azucena O;Chen DC;Sullivan W;Kubby J
活体样品的非均匀折射率引起的空间和时间相关光学像差限制了活体动态成像的分辨率。我们介绍了一种自适应光学显微镜，它使用Shack-Hartmann波前传感器和荧光蛋白质引导星进行实时成像，并采用直接波前传感方法。果蝇胚胎的成像结果表明，它有能力纠正像差，并获得大果蝇胚胎内侧切片的接近衍射极限的图像。GFP-POLO标记的中心体在校正后可以清楚地观察到，但在校正前无法观察到。通过动态像差校正，获得了四维时间推移图像。这些研究还表明，GFP标记的中心体蛋白Polo和CNN是基于自适应光学显微镜的优秀生物指南。
Spatially and temporally dependent optical aberrations induced by the inhomogeneous refractive index of live samples limit the resolution of live dynamic imaging. We introduce an adaptive optical microscope with a direct wavefront sensing method using a Shack-Hartmann wavefront sensor and fluorescent protein guide-stars for live imaging. The results of imaging Drosophila embryos demonstrate its ability to correct aberrations and achieve near diffraction limited images of medial sections of large Drosophila embryos. GFP-polo labeled centrosomes can be observed clearly after correction but cannot be observed before correction. Four dimensional time lapse images are achieved with the correction of dynamic aberrations. These studies also demonstrate that the GFP-tagged centrosome proteins, Polo and Cnn, serve as excellent biological guide-stars for adaptive optics based microscopy.