Quantification of mitotic chromosome alignment

Abstract

The alignment of chromosomes during metaphase is a hallmark of mitosis. For this reason, chromosome alignment has served as an informative functional assay for evaluating mitotic fidelity. The common approach of quantifying the number of mitotic cells with unaligned chromosomes within a population has led to the identification of many proteins required for this conserved process. However, more sensitive assays are now required to dissect the complex molecular control of chromosome alignment. In this chapter, we describe a microscopy-based method for objectively quantifying the distribution of fluorescently labeled chromosomes within the mitotic spindle that can be used to evaluate the extent of chromosome alignment within individual mitotic cells.