Key Words: confocal, fluorescence, live cell imaging, spectral imaging, time-lapse, 3-D reconstruction, voxel
Definition:Imaging that allows the capture of extra dimensions of data over and above that of a simple 2-dimensional image (for example, X, Y, X, time, wavelength, multipoint, sound)
TECHNOLOGY:
Developments in optical microscopy have allowed multiple data to be collected during imaging. The development of confocal imaging, for example, has allowed specimens to be imaged in three dimensions (X, Y, Z) and, with appropriate software, to be rotated to provide an all- round view of the specimen. With the advent of non-toxic fluorophores and time-lapse imaging, 3-D imaging can be extended to include the fourth dimension of time. Spectral imaging also allows multiple wavelengths of light to be simultaneously captured and separated to capture data from multiple fluorescent labels. Fluorophores may also be analyzed in terms of excited state lifetimes rather than emission wavelengths to provide additional information on molecular environments and the physiological state of cells. Every voxel in an image carries information for analysis. A microscope with motorized stage, in addition, enables automatic capture of images at multiple points (X, Y, Z) in the specimen during an imaging session. Multidimensional data capture and analysis requires appropriate software.
APPLICATIONS:
Multidimensional imaging is particularly appropriate to live cell imaging, where simultaneous and continuous data acquisition provides improved information on cell physiology and function (rather than interpolating information from 2-D images separated in time and possibly from different specimens).
MICROSCOPE CONFIGURATION:
Multidimensional imaging can be applied to a variety of imaging systems - depending on the type of date being captured. Confocal systems or multiphoton systems and motorized microscopes provide the greatest scope for multidimensional data capture.
RECOMMENDED SYSTEM:
Eclipse Ti-E/Ti-PFS with environmental control and confocal system (especially the C1si or LiveScan Sweptfield system). The LiveScan Sweptfield system incorporates NIS-Elements for multi-dimensional imaging (X,Y,Z, wavelength, time, multipoint) with support for capture, display, peripheral device control, and data management. It also offers sophisticated image processing features, such as an extremely powerful deconvolution module.