Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, Virginia, USA.
Nat Protoc. 2015 Nov;10(11):1679-96. doi: 10.1038/nprot.2015.111. Epub 2015 Oct 1.
Light-sheet microscopy is a powerful method for imaging the development and function of complex biological systems at high spatiotemporal resolution and over long time scales. Such experiments typically generate terabytes of multidimensional image data, and thus they demand efficient computational solutions for data management, processing and analysis. We present protocols and software to tackle these steps, focusing on the imaging-based study of animal development. Our protocols facilitate (i) high-speed lossless data compression and content-based multiview image fusion optimized for multicore CPU architectures, reducing image data size 30-500-fold; (ii) automated large-scale cell tracking and segmentation; and (iii) visualization, editing and annotation of multiterabyte image data and cell-lineage reconstructions with tens of millions of data points. These software modules are open source. They provide high data throughput using a single computer workstation and are readily applicable to a wide spectrum of biological model systems.
光片显微镜是一种强大的方法,可以在高时空分辨率和长时间尺度上对复杂生物系统的发育和功能进行成像。此类实验通常会生成数太字节的多维图像数据,因此需要有效的计算解决方案来进行数据管理、处理和分析。我们提出了用于解决这些步骤的协议和软件,重点是基于成像的动物发育研究。我们的协议有助于:(i) 针对多核 CPU 架构进行高速无损数据压缩和基于内容的多视图图像融合,将图像数据大小减少 30-500 倍;(ii) 自动进行大规模细胞跟踪和分割;以及 (iii) 可视化、编辑和注释数太字节的图像数据以及具有数千万个数据点的细胞谱系重建。这些软件模块是开源的。它们使用单个计算机工作站提供高数据吞吐量,并且易于应用于广泛的生物模型系统。