Tomographic sensing and localization of fluorescently labeled circulating cells in mice in vivo.
Tomographic sensing and localization of fluorescently labeled circulating cells in mice in vivo.
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DOI：
10.1088/0031-9155/57/14/4627
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发表时间：
2012-07-21
影响因子：
3.5
通讯作者：
Niedre M
中科院分区：
文献类型：
--
作者：
Zettergren E;Swamy T;Runnels J;Lin CP;Niedre M
感知和计数小动物体内特定类型的循环细胞是许多生物医学研究领域的重要问题。基于显微镜的活体荧光流式细胞术方法已经开发出来，但这些方法通常仅限于对非常小的血容量进行采样，因此非常罕见的循环细胞可能会逃脱检测。最近，我们描述了一种‘漫荧光流式细胞仪’(DFFC)的发展，它允许对更大的血管进行采样，从而可以在小鼠的后肢、前肢或尾巴中循环血容量。在这项工作中，我们通过开发和验证一种方法来扩展这一概念，该方法可以在模拟光流体模和小鼠肢体的组织横截面上对循环荧光标记细胞进行断层扫描定位。这是使用两个调制光源和六个光纤耦合探测器阵列实现的，这些探测器允许以10赫兹的速度快速、高灵敏度地获取完整的断层扫描数据集。利用蒙特卡罗光传播模型和随机代数重建技术将这些图像重建成二维横截面图像。我们能够获得样本横截面上移动细胞的连续图像，精度为0.5 mm或更高。我们首先在具有多达四个流动通道的模拟肢体的光流光子中展示了这一概念，然后在带有荧光标记的多发性骨髓瘤细胞的小鼠的尾巴中展示了这一概念。这种方法提高了我们的DFFC仪器的整体诊断效用。
Sensing and enumeration of specific types of circulating cells in small animals is an important problem in many areas of biomedical research. Microscopy-based fluorescence in vivo flow cytometry methods have been developed previously, but these are typically limited to sampling of very small blood volumes, so that very rare circulating cells may escape detection. Recently, we described the development of a ‘diffuse fluorescence flow cytometer’ (DFFC) that allows sampling of much larger blood vessels and therefore circulating blood volumes in the hindlimb, forelimb or tail of a mouse. In this work, we extend this concept by developing and validating a method to tomographically localize circulating fluorescently labeled cells in the cross section of a tissue simulating optical flow phantom and mouse limb. This was achieved using two modulated light sources and an array of six fiber-coupled detectors that allowed rapid, high-sensitivity acquisition of full tomographic data sets at 10 Hz. These were reconstructed into two-dimensional cross-sectional images using Monte Carlo models of light propagation and the randomized algebraic reconstruction technique. We were able to obtain continuous images of moving cells in the sample cross section with 0.5 mm accuracy or better. We first demonstrated this concept in limb-mimicking optical flow photons with up to four flow channels, and then in the tails of mice with fluorescently labeled multiple myeloma cells. This approach increases the overall diagnostic utility of our DFFC instrument.
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