The mode of acquisition defines the pixel image data. The meaning of a 2D-image recorded from a digital microscope imaging system varies depending on how it is collected. Almost all of the different modes in (a) and (b) can be combined to analyze cell structure and behavior. All of the parameters and configurations must be somehow recorded for the interpretation of the pixel data in an image. (a) The spatial, spectral and temporal context of an image is used to generate more information about the cell under study. Changing stage position, focus, spectral range or time of imaging all expand the meaning of an image. Modified from . (b) The two aspects of the image data collection that define the pixel data. A variety of methods are used to generate contrast in modern biological imaging. In addition, the imaging method used to record the data also has meaning.