Live-cell imaging chambers are used in a wide range of cell biology research. Recently, chambers capable of taking high-resolution and time-lapse images of live cells have been developed and become commercially available. However, since most of these chambers are designed to maintain a thermally stable environment for the cells under study, it is usually very difficult to use them to study temperature-dependent cellular events. Here we report the development of a chamber that is able to be used for the continuously monitoring of live neurons under most commercially available up-right epi-fluorescence and confocal microscopes and in which the temperature and composition of the medium surrounding the neurons can be changed rapidly and reversibly. This live-cell observation chamber has been used successfully with cultured rat hippocampal neurons to study temperature-dependent changes in the dynamics of the microtubule cytoskeleton using Fluorescence Recovery after Photobleaching (FRAP) together with the localization of α-tubulin in the dendritic spines. The caffeine-induced exit of α-tubulin from the dendritic spines of neurons could also be monitored using this chamber. The success of these observations demonstrates the usefulness and applicability of the live-cell observation chamber described here to a wide range of cell biology experiments.

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