An observation chamber for studying temperature-dependent and drug-induced events in live neurons using fluorescence microscopy

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, because most of these chambers are designed to maintain a thermally s...

Full description

Saved in:
Bibliographic Details
Published in:Analytical biochemistry 2009-03, Vol.386 (1), p.105-112
Main Authors: Cheng, Wen-Yao, Hsu, Wei-Lun, Cheng, Hui-Hsuan, Huang, Zu-Han, Chang, Yen-Chung
Format: Article
Language:English
Subjects:
Animals
Cells, Cultured
Chamber
Cytoskeleton - ultrastructure
Dendritic Spines - ultrastructure
Fluorescence microscopy
Fluorescence Recovery After Photobleaching
Hippocampus - cytology
Live cell
Methods
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Neurons - drug effects
Neurons - ultrastructure
Pharmaceutical Preparations
Rats
Research Design
Temperature
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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, because 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 continuous monitoring of live neurons under most commercially available upright epifluorescence 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 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.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2008.12.004