Live Cell Imaging: Studying NF-kB Signaling Dynamics Using the Operetta System

High content imaging provides a rapid and flexible tool to quantitatively study the intracellular localization of fluorescent markers on a single cell basis. In this application note from PerkinElmer a live cell imaging approach is presented for studying the nuclear translocation of NF-κB using the Operetta® High Content Imaging System. In contrast to fixed endpoint assays, this allows the negative feedback of NF-κB signaling, which results from NF-κB induced expression of IκB, to be analyzed. The use of Halo-tagged proteins on the Operetta system allows sensitive and flexible live cell assays to be performed for diverse high content screening applications.