期刊
IMMUNOLOGY AND CELL BIOLOGY
卷 87, 期 2, 页码 154-158出版社
WILEY
DOI: 10.1038/icb.2008.79
关键词
antigen presentation; imaging; lymphocytes; microfabrication; T cells; thymocytes
资金
- Australian Research Council,
- National Health and Medical Research Council
- Human Frontier Science Program
With new imaging technologies and fluorescent probes, live imaging of cells in vitro has revolutionized many aspects of cell biology. A key goal now is to develop systems to optimize in vitro imaging, which do not compromise the physiological relevance of the study. We have developed a methodology that contains non-adherent cells within the field of view. 'Cell paddocks' are created by generating an array of microgrids using polydimethylsiloxane. Each microgrid is up to 250 x 250 mu m(2) with a height of 60 mu m. Overlayed cells settle into the grids and the walls restrict their lateral movement, but a contiguous supply of medium between neighboring microgrids facilitates the exchange of cytokines and growth factors. This allows culture over at least 6 days with no impact upon viability and proliferation. Adaptations of the microgrids have enabled imaging and tracking of lymphocyte division through multiple generations of long-term interactions between T lymphocytes and dendritic cells, and of thymocyte stromal cell interactions.
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