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首页> 外文会议>Conference on Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing Ⅷ Jan 21-22, 2001, San Jose, USA >Design of a Confocal Fluorescence Spectrometer for Transdermal Observation of Green Fluorescent Protein Expression in Cartilage
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Design of a Confocal Fluorescence Spectrometer for Transdermal Observation of Green Fluorescent Protein Expression in Cartilage

机译：用于透皮观察软骨中绿色荧光蛋白表达的共聚焦荧光光谱仪的设计

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摘要

We have developed a noninvasive confocal fluorescence spectrometer to measure in vivo fluorescence spectra in mice. Our motivation is the study of the healing process of wounded cartilage in the murine model. The spectrometer measures the expression of green fluorescent protein (GFP) which is linked to the promoter of type II collagen as a marker for the expression of the chondrocyte phenotype. The confocal system uses an argon ion laser (488 nm) to excite fluorescence in a confocal volume about 200 um below the skin surface in the cartilage of the xyphoid process. In vitro studies showed how the refractive mismatch at the surface boundary and the light scattering of the tissue affect the depth and size of the confocal volume. A thin fluorescent layer was detected through a 130 um tissue phantom (murine skin) by moving the confocal volume axially through the phantom. The presence of skin increased the axial full width at half maximum of the confocal response from 7 um to 72 um. In conclusion, we achieve a low-spatial-resolution confocal spectrometer, which yields a fluorescence spectrum that maximizes the GFP fluorescence of cartilage and minimizes the skin autofluorescence.
机译：我们已经开发了一种非侵入式共聚焦荧光光谱仪来测量小鼠体内的荧光光谱。我们的动机是在鼠模型中研究受伤软骨的愈合过程。光谱仪测量绿色荧光蛋白（GFP）的表达，该蛋白与II型胶原蛋白的启动子相连，作为软骨细胞表型表达的标记。共聚焦系统使用氩离子激光（488 nm）来激发荧光，使共焦体积中的胶体突软骨中皮肤表面以下200 um处发出荧光。体外研究表明，在表面边界处的折射失配和组织的光散射如何影响共聚焦体积的深度和大小。通过将共焦体积轴向移动通过幻影，可以检测到穿过130 um组织幻影（小鼠皮肤）的薄荧光层。皮肤的存在将共聚焦响应的一半处的轴向全宽从7 um增加到72 um。总之，我们实现了一种低空间分辨率共聚焦光谱仪，该光谱仪产生的荧光光谱可使软骨的GFP荧光最大化，并使皮肤的自体荧光最小。

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