A method for prolonged imaging of motile lymphocytes

作者:Day Daniel; Pham Kim; Ludford Menting Mandy J; Oliaro Jane; Izon David; Russell Sarah M*; Gu Min
来源:Immunology and Cell Biology, 2009, 87(2): 154-158.
DOI:10.1038/icb.2008.79

摘要

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.

  • 出版日期2009-2