Showing their true colors: a practical approach to volume rendering from serial sections.
Showing their true colors: a practical approach to volume rendering from serial sections.
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DOI：
10.1186/1471-213x-10-41
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发表时间：
2010-04-21
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通讯作者：
Metscher BD
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文献类型：
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作者：
Handschuh S;Schwaha T;Metscher BD
与更现代的成像方法相比，传统的光学显微镜仍然提供了一系列关于对比度选项，可访问的标本大小和分辨率的实质性优势。当前，尤其是断层摄影图像数据最通常使用体绘制（volume rendering）在三维中可视化。到目前为止，这种方法很少被应用于从连续切片中获取的图像堆栈，而表面渲染仍然是三维呈现此类数据集的最流行方法。本研究的目的是开发连续切片图像堆栈体积渲染的标准协议，同时保留光学显微镜的优点，例如分辨率和颜色信息。在这里，我们提供了一套协议，用于获取高分辨率的3D图像的不同的显微镜样品通过体绘制的基础上，使用3D重建软件阿米拉（Visage成像公司）的连续光显微镜切片。我们克服了一些技术障碍，并表明这些渲染的质量和分辨率与使用其他方法的3D可视化相当。这种用于以全色可视化3D微观形态的实用方法利用了光学显微镜的亚微米分辨率和组织学染色的特异性，通过结合传统的组织学切片技术、数字图像采集、三维图像过滤以及3D图像处理和可视化技术。我们表明，这种方法可以产生“真正的”彩色高分辨率的三维视图的组织以及细胞和亚细胞结构，从而代表了一个强大的工具，形态，发育和比较研究。我们的结论是，所提出的方法填补了一个重要的空白，在该领域的显微解剖三维成像和可视化方法相结合的组织分辨率和分化的细节与三维渲染的整个组织样本。我们证明了选定的无脊椎动物和脊椎动物标本的方法，并建议重新调查的历史连续切片材料可被视为一个特殊的好处。
In comparison to more modern imaging methods, conventional light microscopy still offers a range of substantial advantages with regard to contrast options, accessible specimen size, and resolution. Currently, tomographic image data in particular is most commonly visualized in three dimensions using volume rendering. To date, this method has only very rarely been applied to image stacks taken from serial sections, whereas surface rendering is still the most prevalent method for presenting such data sets three-dimensionally. The aim of this study was to develop standard protocols for volume rendering of image stacks of serial sections, while retaining the benefits of light microscopy such as resolution and color information. Here we provide a set of protocols for acquiring high-resolution 3D images of diverse microscopic samples through volume rendering based on serial light microscopical sections using the 3D reconstruction software Amira (Visage Imaging Inc.). We overcome several technical obstacles and show that these renderings are comparable in quality and resolution to 3D visualizations using other methods. This practical approach for visualizing 3D micro-morphology in full color takes advantage of both the sub-micron resolution of light microscopy and the specificity of histological stains, by combining conventional histological sectioning techniques, digital image acquisition, three-dimensional image filtering, and 3D image manipulation and visualization technologies. We show that this method can yield "true"-colored high-resolution 3D views of tissues as well as cellular and sub-cellular structures and thus represents a powerful tool for morphological, developmental, and comparative investigations. We conclude that the presented approach fills an important gap in the field of micro-anatomical 3D imaging and visualization methods by combining histological resolution and differentiation of details with 3D rendering of whole tissue samples. We demonstrate the method on selected invertebrate and vertebrate specimens, and propose that reinvestigation of historical serial section material may be regarded as a special benefit.