Automated Identification and Localization of Hematopoietic Stem Cells in 3D Intravital Microscopy Data.
Automated Identification and Localization of Hematopoietic Stem Cells in 3D Intravital Microscopy Data.
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DOI：
10.1016/j.stemcr.2015.05.017
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发表时间：
2015-07-14
影响因子：
5.9
通讯作者：
Lo Celso C
中科院分区：
文献类型：
--
作者：
Khorshed RA;Hawkins ED;Duarte D;Scott MK;Akinduro OA;Rashidi NM;Spitaler M;Lo Celso C
使用活体显微镜测量骨髓微环境内造血干细胞（HSC）的三维（3D）定位是一个迅速扩展的研究主题。这种方法是理解HSC-小生境相互作用细节的关键，这对适当的干细胞功能至关重要。由于骨髓的复杂组织结构以及随着成像深度的增加散射和信号损失的逐渐引入，没有现成的软件来处理数据的有效分割和无偏分析。为了解决这个问题，我们开发了一种自动化的图像分析工具，可以简化和简化3D HSC微环境图像的生物学解释。该算法识别HSC并测量其相对于周围成骨细胞和骨胶原的定位。我们在这里证明了所提出的方法的有效性，一致性和准确性相比，目前的手动分析和其更广泛的适用性，以分析其他3D骨髓成分。一种新的工具允许HSC及其小生境的自动3D图像分析它执行异质HSC和骨髓成分的自动分割该工具识别真实的HSC并消除假阳性信号证明了HSC到最近的成骨细胞/骨的3D距离测量Lo Celso，Khorshed及其同事描述了一种高度灵活的自动计算方法，该方法允许识别HSC，成骨细胞，和骨胶原蛋白，并测量它们在小鼠骨髓活体显微镜图像中的3D相互定位。该方法适用于研究活组织内感兴趣的单细胞与任何其他标记细胞或结构之间的相互作用。
Measuring three-dimensional (3D) localization of hematopoietic stem cells (HSCs) within the bone marrow microenvironment using intravital microscopy is a rapidly expanding research theme. This approach holds the key to understanding the detail of HSC-niche interactions, which are critical for appropriate stem cell function. Due to the complex tissue architecture of the bone marrow and to the progressive introduction of scattering and signal loss at increasing imaging depths, there is no ready-made software to handle efficient segmentation and unbiased analysis of the data. To address this, we developed an automated image analysis tool that simplifies and standardizes the biological interpretation of 3D HSC microenvironment images. The algorithm identifies HSCs and measures their localization relative to surrounding osteoblast cells and bone collagen. We demonstrate here the effectiveness, consistency, and accuracy of the proposed approach compared to current manual analysis and its wider applicability to analyze other 3D bone marrow components. A new tool allows automated 3D image analysis of HSCs and their niche It performs automated segmentation of heterogeneous HSCs and bone marrow components This tool identifies real HSCs and eliminates false-positive signals 3D distance measurements of HSC to the nearest osteoblast/bone are demonstrated Lo Celso, Khorshed, and colleagues describe a highly flexible, automated computational method that allows identification of HSCs, osteoblast cells, and bone collagen and measurement of their 3D reciprocal localization in intravital microscopy images of mouse bone marrow. This approach is applicable to study the interaction between single cells of interest and any other labeled cells, or structures, within live tissues.