Mansfield James R, Hoyt Clifford, Levenson Richard M
Cambridge Research & Instrumentation, Woburn, Massachusetts, USA.
Curr Protoc Mol Biol. 2008 Oct;Chapter 14:Unit 14.19. doi: 10.1002/0471142727.mb1419s84.
Combining images taken with light of specific wavelengths can dramatically enhance light-microscopic images. This technology is enabled by the availability of programmable filters that can be set to transmit light only of particular wavelengths. Spectral imaging technologies have become an important part of microscopy, and are particularly useful for analyzing samples that have been labeled with multiple (two or more) molecular markers. The most commonly used methodology for separating the markers from each other is linear unmixing, which results in a quantitative image of the location and amount of each marker present in the sample. The very complexity of these multilabel samples requires a high degree of sophistication in methods to visualize the results of unmixing. This article describes a wide range of useful visualization tools designed to better enable discrimination of different features in multilabeled tissue or cell samples. These commercially available tools can be attached to the standard laboratory light microscope to significantly enhance the power of light microscopy.
结合特定波长光拍摄的图像可以显著增强光学显微镜图像。这项技术得益于可编程滤光片的应用，这些滤光片可以设置为只透射特定波长的光。光谱成像技术已成为显微镜学的重要组成部分，对于分析用多种（两种或更多）分子标记物标记的样本特别有用。最常用的将标记物彼此分离的方法是线性解混，这会生成样本中每个标记物的位置和数量的定量图像。这些多标记样本的复杂性极高，因此需要高度精密的方法来可视化解混结果。本文介绍了一系列有用的可视化工具，旨在更好地辨别多标记组织或细胞样本中的不同特征。这些可商购的工具可以连接到标准实验室光学显微镜上，以显著增强光学显微镜的功能。
