Live Cell Microscopy Imaging

Presenter: Simon Watkins

Abstract

In the post-genomic era of biomedical research understanding the functionality of molecules at the cellular and subcellular level in living systems will become predominant. In this era we must move beyond static “snapshots” of the cellular state to an understanding of the biology of cells over time and in 3-dimentional- space. Within the cellular environment it is expected that we will be able to study the expression, the functional role(s) and interactions of multiple unique molecules concurrently. Furthermore, it will be desirable to determine the effects of these molecules on cell development, organization and fate over extended periods of time. To perform these types of studies it is necessary to develop new methodologies that will allow multiparametric analysis of cells while maintaining their functional viability. In the past this goal would have been extraordinarily difficult to achieve. However, developments in optical and computational technology have empowered modern microscopists to undertake these previously forbidding tasks. This two session workshop will discuss live cell imaging tools, the expectations of the technology and limitations of optical tools within the context of current scientific efforts principally focusing on the use of fluorescent proteins and ratiometric tools in live cell methodologies

1. The first session in this workshop will focus on:
• optical principals
• principals of fluorescence
• detector and microscope design
2. The second session will focus on actual methods:
• Multicolor fluorescence,
• FRET
• TIRF
• GFP
• Live cell confocal