Live Cell Imaging: Tips and Tools

Abstract

Imaging of living cells and tissue is now common in many fields of the life sciences and the physical sciences as well. It is critical when performing such experiments that cell viability is at the forefront of any measurement to ensure the physiological/biological processes under investigation are not altered in any way. The main cause of cyto-toxicity in living cells is due to the reaction of free radical species, generated during the excitation of fluorescent proteins, or dye molecules, with surrounding molecules. The amount of photo-toxicity is directly related to the amount of light energy put into the system, thus, it is critical to minimize light exposure as much as possible. This commentary discusses how to set up a suitable environment on the microscope stage to maintain living cells. While the main focus, is on general and imaging platform specific ways to minimize light exposure during live-cell imaging. Reducing the power of excitation light, maximizing efficiency of the optical path, and optimizing detector settings are all ways light exposure can be minimized. Brief suggestions for useful microscope accessories as well as available fluorescence tools are also presented. Finally, a flow chart is offered to assist readers in choosing the appropriate imaging platform for their experimental system.