High resolution multimode light microscopy of cell migration: long-term imaging and analysis.

Abstract

Cell migration is a multi-step process that involves sequential changes in the cytoskeleton, cell-substrate adhesion and components of the extracellular matrix. In multicellular organisms, directional cell migration is important for normal development, wound healing and immune responses and contributes to disease states such as tumor formation and metastasis. Many cells such as fibroblasts migrate as individuals while others, such as keratinocytes, move as groups or sheets of cells.In this chapter, we use human oral keratinocytes (OKF6/TERT-2) to illustrate the complex patterns of cell migration and its regulation. In culture, sheets of keratinocytes migrate and respond to human pathogens such as Candida albicans. The dynamic changes of the cytoskeleton, cell-cell and cell-substrate interactions that change during an infection for example require observation over long periods of time in order to identify the spatio-temporal coordinated regulation of the cytoskeleton and its associated components as well as the signaling pathways that control them.Microscopic techniques for long-term live cell observation and analysis of cell migration require high-resolution imaging systems that maintain perfect focus and optimal growth conditions (temperature, CO(2)) for cells. We describe two multimode digital imaging systems (VEC-DIC and BioStation IM), both with wide-field epifluorescence and transmitted light objectives for long-term time-lapse imaging and motion analysis.