Automatic cell counting in vivo in the larval nervous system of Drosophila.
Automatic cell counting in vivo in the larval nervous system of Drosophila.
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DOI：
10.1111/j.1365-2818.2012.03608.x
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发表时间：
2012-05
影响因子：
2
通讯作者：
Hidalgo A
中科院分区：
文献类型：
--
作者：
Forero MG;Kato K;Hidalgo A
细胞的识别和计数对于测试生物学假设是必要的，例如神经系统形成、疾病、退化、损伤和再生，但手动计数耗时、繁琐，并且容易产生偏差。果蝇是一种广泛用于分析基因功能的模式生物，大多数研究是在完整的动物或整个器官中进行，而不是在细胞培养物中进行。对基因功能的推断需要从这些样本类型中了解细胞计数。图像处理和模式识别技术是自动化细胞计数的合适工具。然而，对果蝇中的细胞进行计数是一项复杂的任务：免疫组织化学标记和发育阶段的变化导致图像具有非常不同的特性，使得识别真正的细胞具有挑战性。在这里，我们提出了一种通过共焦显微镜系列光学切片自动计数 3D 幼虫胶质细胞的技术。结合局部离群值阈值和圆顶来查找细胞。从数据集中提取的形状描述符用于表征细胞并避免过度分割。采用形态算子来划分否则可能会被遗漏的细胞。该方法准确且非常快速，并且平等且客观地对待所有样本，使所有数据在样本之间具有可比性。我们的方法也适用于识别用其他核标记物标记的细胞和小鼠组织切片中的细胞。
Identification and counting of cells is necessary to test biological hypotheses, for instance of nervous system formation, disease, degeneration, injury and regeneration, but manual counting is time-consuming, tedious, and subject to bias. The fruit-fly Drosophila is a widely used model organism to analyse gene function, and most research is carried out in the intact animal or in whole organs, rather than in cell culture. Inferences on gene function require that cell counts are known from these sample types. Image processing and pattern recognition techniques are appropriate tools to automate cell counting. However, counting cells in Drosophila is a complex task: variations in immunohistochemical markers and developmental stages result in images of very different properties, rendering it challenging to identify true cells. Here, we present a technique for counting automatically larval glial cells in 3D, from confocal microscopy serial optical sections. Local outlier thresholding and domes are combined to find the cells. Shape descriptors extracted from a data set are used to characterise cells and avoid over-segmentation. Morphological operators are employed to divide cells that could otherwise be missed. The method is accurate and very fast, and treats all samples equally and objectively, rendering all data comparable across specimens. Our method is also applicable to identify cells labelled with other nuclear markers and in sections of mouse tissues.