Culturing Cells on the Microscope Stage

Abstract

This chapter discusses the microscope culture system and cell culture chambers. Keeping live, healthy cells on the microscope is crucial for many types of experiments, in particular, fluorescence microscopy of dynamic processes. It is found that although cultured cells may be observed for a short period of time without any special device, a culture system is usually necessary for observations which last for more than 10 min. The major requirements for maintaining the cell are temperature, pH, osmolarity, and nutrients. Cultured mammalian cells should generally be maintained at a temperature close to 37°C. It is observed that with the exception of temperature-sensitive mutants, most cells can tolerate a slightly lower temperature, as well as slight variations in the temperature. The required performance of a microscope culture system depends on the period of observation, the nature of the experiment, and the property of the cell. On the one hand, for short-term observations, the best design may be simply a sealed cover slip. The most sophisticated chambers incorporate mechanisms for the control of various parameters, without the need of a separate microscope incubator. It is found that for open chambers, even with an oil overlay, the pH of the bicarbonate buffer must be maintained by a continuous supply of CO2. © 1989, Academic Press Inc. All rights reserved.