Abstract
The modern microscope has advanced so far beyond what Leeuwenhoek invented, it's doubtful he would even recognize it as a microscope. Modern microscopes, or more acutely, imaging workstations, are an integrated compilation of optoelectronic components. It is now possible to produce much more than simple images using these systems. Quantitation of various metrics are now possible with these advanced instruments. With the advent of fluorescently-tagged proteins/organelles, it is possible to actually quantify sub-cellular amounts of these transgenically produced or exogenously labeled macromolecules/complexes. Spectral un-mixing of overlapping signals from within a single image (fluorescent and absorbent) has blurred distinction between imaging and spectroscopy. It is now possible to get chemical data from techniques such as unmixing in addition to more conventional modes such as polarized light microscopy. The ability to determine if two macromolecules are precisely co-localized (< 100 A apart) is now routinely accomplished in both living and fixed preparations with Fluorescence Resonance Energy Transfer (FRET). The latest technological advances has been achieving resolution beyond Abbe's predicted limits. This has been accomplished both through hardware and software improvements, such as Stimulated Emission Depletion (STED), and Structured Illumination Microscopy (SIM) to name a couple.
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