While providing indispensable insight into immune processes, the live cell imaging of immune cells poses unique challenges due to their non-adherent nature. Suspension cells such as immune cells can easily move away from the FOV (field of view) during imaging by perturbation such as reagent introduction, often necessitating the use of surface coating to artificially immobilize cells. In addition, unlike adherent cells, interacting suspension cells form aggregates, making it very difficult to track and analyze individual cells during imaging. Here we describe the novel, microfluidic-based dynamic live cell imaging platform CellASIC® ONIX2, where microenvironmental parameters such as the perfusion of media/reagent, temperature, and gas compositions can be precisely controlled on-demand via software during the entire imaging experiment. Unlike most live imaging platforms merely aiming to maintain temperature and gas at certain levels, CellASIC® ONIX2 platform’s ability to control fluidics and perfusion offers true on-demand control of cellular microenvironment during live cell imaging, and its standard plate form factor can instantly upgrade any existing inverted microscope into a fully capable live cell imaging platform. Overview of applications covering hypoxia, apoptosis, migration, and suspension immune cell imaging will be presented. Specific emphasis will be given to microfluidic designs targeted for use with different cell types as well as fluorescent probes for live cell imaging. Any scientists planning to start live cell imaging experiments, as well as experienced imaging scientists wanting to broaden their applications will benefit from this session.
Learning Objectives:
1. Evaluate the advantages and limitations of current live cell imaging platforms
2. Discuss the emerging field of microfluidic based dynamic imaging
3. Review current state of the art regarding microfluidics based dynamic imaging