Measuring secretory membrane traffic: a quantitative fluorescence microscopy approach

V Starkuviene, A Seitz, H Erfle, R Pepperkok - Membrane Trafficking, 2008 - Springer
Membrane Trafficking, 2008Springer
In this chapter the authors describe automated imaging methods to quantify the transport
rates of transmembrane as well as soluble cargo, and to evaluate the integrity of the Golgi
complex. The quantification of cargo transport rates serves as an example of fluorescence
intensity-based assays, the quantification of the Golgi complex integrity—as an example of
morphology-based assays. These quantitative assays could be applied for single
experiments as well as for middle-and high-throughput screening approaches. Each of …
Summary
In this chapter the authors describe automated imaging methods to quantify the transport rates of transmembrane as well as soluble cargo, and to evaluate the integrity of the Golgi complex. The quantification of cargo transport rates serves as an example of fluorescence intensity-based assays, the quantification of the Golgi complex integrity—as an example of morphology-based assays. These quantitative assays could be applied for single experiments as well as for middle- and high-throughput screening approaches. Each of these assays can be used to appreciate effects caused by gene silencing by RNAi, cDNA overexpression or application of chemical compounds. For each assay the authors discuss protocols for sample preparation, parameters for automated image acquisition, strategies of image analysis, and data quantification.
Springer