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首页> 外文会议>Conference on Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II; 20040127-20040128; San Jose,CA; US >Confocal and two-photon imaging in cartilage: expression patterns of Filamin A and B
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Confocal and two-photon imaging in cartilage: expression patterns of Filamin A and B

机译：软骨共焦和双光子成像：丝蛋白A和B的表达模式

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摘要

Optical imaging in cartilage is challenging due to the high levels of intra- and inter-cellular autofluorescence. We report here on high-resolution confocal and two-photon imaging of endogenous fluorescence of cartilage and of exogenous fluorescence of filamin A and B protein markers. Confocal laser scanning microscopy offers the advantage of quasi-theoretical spatial resolution and minimizes the autofluorescence contribution by eliminating the out-of-focus light. In non-labeled cartilage, we observe mostly intracellular autofluorescence that, due to the uniform distribution within the cell, can be further effectively minimized by careful choice of experimental parameters. The fluorescence of the exogenous markers AlexaFluor 488 and AlexaFluor 568 labeling Filamins A and B, respectively, could also be detected and quantitated using this procedure, revealing topologically different expression levels of filamin A and B proteins in the cartilage growth plate. Two-photon excited fluorescence imaging yielded further resolution improvements and structural and functional information.
机译：由于细胞内和细胞间自发荧光水平高，软骨中的光学成像具有挑战性。我们在这里报告关于软骨的内源性荧光以及丝素A和B蛋白标记的外源荧光的高分辨率共聚焦和双光子成像。共聚焦激光扫描显微镜具有准理论空间分辨率的优势，并且通过消除散焦光而将自发荧光的影响降至最低。在未标记的软骨中，我们观察到大多数细胞内自发荧光，由于细胞内的均匀分布，可以通过仔细选择实验参数来进一步有效地使之最小化。还可以使用此程序检测和定量标记Filamins A和B的外源标记AlexaFluor 488和AlexaFluor 568的荧光，揭示了软骨生长平板中filamin A和B蛋白的拓扑结构不同的表达水平。双光子激发荧光成像可进一步提高分辨率，并获得结构和功能信息。

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