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Cell Culture Environment on the Microscope
Cells will only behave naturally when they are cultured under the specific conditions of their biological environment. In mammals, the most prominent conditions are a temperature of 37°C, a pH of 7.4 that is controlled by a bicarbonate buffer at a 5 % CO2 concentration, and constant concentrations of salts and nutrients. Depending on the location inside the mammalian body, the oxygen concentration is another important parameter to control. It can range from 14% in the lung tissue to 0% in solid tumors.
In order to achieve biologically relevant results, it is crucial to maintain these conditions on the microscope stage during live cell imaging experiments.
Many cellular processes can be visualized directly on the microscope. Some of these in vitro assays, like rolling adhesion, chemotaxis, or tube formation assays, don’t even require staining. These assays can easily be performed with the simple phase contrast technique. However, more structural details and molecular interaction can be revealed by fluorescence microscopy, using reporter proteins like GFP. Highest magnification microscopy (e.g., TIRF) focuses on single molecule dynamics, such as the cellular F-Actin cytoskeleton. LifeAct, ibidi’s F-actin marker, is a prominent example of that.