Ozcan Burcin, Negi Pooran, Laezza Fernanda, Papadakis Manos, Labate Demetrio
Dept. of Mathematics, University of Houston, Houston, Texas, United States of America.
Dept. of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston, Texas, United States of America.
PLoS One. 2015 Apr 8;10(4):e0121886. doi: 10.1371/journal.pone.0121886. eCollection 2015.
Automated identification of the primary components of a neuron and extraction of its sub-cellular features are essential steps in many quantitative studies of neuronal networks. The focus of this paper is the development of an algorithm for the automated detection of the location and morphology of somas in confocal images of neuronal network cultures. This problem is motivated by applications in high-content screenings (HCS), where the extraction of multiple morphological features of neurons on large data sets is required. Existing algorithms are not very efficient when applied to the analysis of confocal image stacks of neuronal cultures. In addition to the usual difficulties associated with the processing of fluorescent images, these types of stacks contain a small number of images so that only a small number of pixels are available along the z-direction and it is challenging to apply conventional 3D filters. The algorithm we present in this paper applies a number of innovative ideas from the theory of directional multiscale representations and involves the following steps: (i) image segmentation based on support vector machines with specially designed multiscale filters; (ii) soma extraction and separation of contiguous somas, using a combination of level set method and directional multiscale filters. We also present an approach to extract the soma's surface morphology using the 3D shearlet transform. Extensive numerical experiments show that our algorithms are computationally efficient and highly accurate in segmenting the somas and separating contiguous ones. The algorithms presented in this paper will facilitate the development of a high-throughput quantitative platform for the study of neuronal networks for HCS applications.
自动识别神经元的主要组成部分并提取其亚细胞特征是许多神经网络定量研究中的关键步骤。本文的重点是开发一种算法,用于自动检测神经网络培养物共聚焦图像中胞体的位置和形态。这个问题是由高内涵筛选(HCS)中的应用所推动的,在高内涵筛选中,需要从大数据集中提取神经元的多个形态特征。现有的算法在应用于神经元培养物共聚焦图像堆栈分析时效率不高。除了处理荧光图像通常存在的困难外,这类堆栈包含的图像数量较少,因此沿z方向可用的像素数量也较少,应用传统的3D滤波器具有挑战性。我们在本文中提出的算法应用了方向多尺度表示理论中的一些创新思想,涉及以下步骤:(i)基于支持向量机并使用专门设计的多尺度滤波器进行图像分割;(ii)使用水平集方法和方向多尺度滤波器相结合的方式提取胞体并分离相邻的胞体。我们还提出了一种使用3D剪切波变换提取胞体表面形态的方法。大量数值实验表明,我们的算法在分割胞体和分离相邻胞体方面计算效率高且精度高。本文提出的算法将有助于开发一个用于HCS应用的神经网络研究的高通量定量平台。