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首页> 外文期刊>Biophysical Journal >A blind spot in confocal reflection microscopy: the dependence of fiber brightness on fiber orientation in imaging biopolymer networks.
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A blind spot in confocal reflection microscopy: the dependence of fiber brightness on fiber orientation in imaging biopolymer networks.

机译：共聚焦反射显微镜的一个盲点：成像生物聚合物网络中纤维亮度对纤维取向的依赖性。

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摘要

We investigate the dependence of fiber brightness on three-dimensional fiber orientation when imaging biopolymer networks with confocal reflection microscopy (CRM) and confocal fluorescence microscopy (CFM). We compare image data of fluorescently labeled type I collagen networks concurrently acquired using each imaging modality. For CRM, fiber brightness decreases for more vertically oriented fibers, leaving fibers above approximately 50 degrees from the imaging plane entirely undetected. As a result, the three-dimensional network structure appears aligned with the imaging plane. In contrast, CFM data exhibit little variation of fiber brightness with fiber angle, thus revealing an isotropic collagen network. Consequently, we find that CFM detects almost twice as many fibers as are visible with CRM, thereby yielding more complete structural information for three-dimensional fiber networks. We offer a simple explanation that predicts the detected fiber brightness as a function of fiber orientation in CRM.
机译：当使用共聚焦反射显微镜（CRM）和共聚焦荧光显微镜（CFM）成像生物聚合物网络时，我们研究了纤维亮度对三维纤维取向的依赖性。我们比较使用每种成像方式同时获取的荧光标记的I型胶原蛋白网络的图像数据。对于CRM，对于更多垂直定向的光纤，其光纤亮度会降低，而与成像平面大约50度以上的光纤将完全未被检测到。结果，三维网络结构看起来与成像平面对齐。相反，CFM数据显示纤维亮度随纤维角度的变化很小，因此揭示了各向同性的胶原网络。因此，我们发现CFM检测到的光纤几乎是CRM可见光纤的两倍，从而为三维光纤网络提供了更完整的结构信息。我们提供了一个简单的解释，可以预测检测到的纤维亮度与CRM中纤维取向的关系。

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