1. ¹Centre for Micro-Photonics, Faculty of Engineering and Industrial Sciences, Swinburne University of Technology, Hawthorn, Vic, Australia
2. ²Immune Signalling Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Vic, Australia
3. ³St Vincent's Institute, Fitzroy, Vic, Australia

Correspondence: Dr SM Russell, Immune Signalling Laboratory, Peter MacCallum Cancer Centre, St Andrews Place, East Melbourne, Vic 3002, Australia. E-mail: sarah.russell@petermac.org; D Day, Centre for Micro-Photonics, Swinburne University of Technology, Hawthorn, Vic, Australia. E-mail: dday@swin.edu.au

Received 26 August 2008; Revised 22 September 2008; Accepted 23 September 2008; Published online 4 November 2008.

Abstract

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 250 250 m² with a height of 60 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.