Cordelières Fabrice P, Bolte Susanne
Bordeaux Imaging Center, UMS 3420 CNRS-Université Bordeaux Segalen-US4 INSERM, Pôle d'imagerie photonique, Institut François Magendie, Bordeaux Cedex, France.
Sorbonne Universités-UPMC Univ Paris 06, Institut de Biologie Paris-Seine-CNRS FR 3631, Cellular Imaging Facility, Paris Cedex, France.
Methods Cell Biol. 2014;123:395-408. doi: 10.1016/B978-0-12-420138-5.00021-5.
Multicolor fluorescence microscopy helps to define the local interplay of subcellular components in cell biological experiments. The analysis of spatial coincidence of two or more markers is a first step in investigating the potential interactions of molecular actors. Colocalization studies rely on image preprocessing and further analysis; however, they are limited by optical resolution. Once those limitations are taken into account, characterization might be performed. In this review, we discuss two types of parameters that are aimed at evaluating colocalization, which are indicators and quantifiers. Indicators evaluate signal coincidence over a predefined scale, while quantifiers provide an absolute measurement. As the image is both a collection of intensities and a collection of objects, both approaches are applicable. Most of the available image processing software include various colocalization options; however, guidance for the choice of the appropriate method is rarely proposed. In this review, we provide the reader with a basic description of the available colocalization approaches, proposing a guideline for their use, either alone or in combination.
多色荧光显微镜有助于在细胞生物学实验中确定亚细胞成分的局部相互作用。分析两个或多个标记物的空间重合是研究分子参与者潜在相互作用的第一步。共定位研究依赖于图像预处理和进一步分析；然而，它们受到光学分辨率的限制。一旦考虑到这些限制，就可以进行表征。在本综述中，我们讨论了旨在评估共定位的两类参数，即指标和量化器。指标在预定义的尺度上评估信号重合，而量化器提供绝对测量。由于图像既是强度的集合又是对象的集合，这两种方法都适用。大多数现有的图像处理软件都包括各种共定位选项；然而，很少有人提出选择合适方法的指导。在本综述中，我们向读者提供可用共定位方法的基本描述，提出单独或组合使用这些方法的指导原则。
