The Fluorescence Microscope
Section Overview:
In fluorescence microscopy, wide variations between localized fluorophore concentrations within the specimen, coupled to differences in extinction coefficient and quantum yield from one fluorochrome to another, significantly influence the emission signal produced for a given quantity of excitation intensity. Considering that many specimens contain only minute quantities of fluorescent material in any particular viewfield, these combined factors produce an average level of fluorescence emission that is four to six orders of magnitude less than the excitation intensity.
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Anatomy of the Fluorescence Microscope
Fluorescence microscopes have evolved with speed over the past decade, coupled to equally rapid advances in laser technology, solid-state detectors, interference thin film fabrication, and computer-based image analysis.
Fluorescence Microscopy with Transmitted Light
Discussed in this section are the various aspects of transmitted fluorescence illumination and equipment configurations needed to perform this type of microscopy.
Introduction to Laser Scanning Microscopes
Learn all about laser scanning microscopes, including their advantages, disadvantages, and the types of images they can produce.
Fluorescence Microscope Schematic Diagrams
Olympus Upright Microscope
Olympus Upright epi-fluorescence microscope is equipped with a vertical illuminator that contains a turret of filter cubes and a fluorescence excitation light source.
Olympus Inverted Microscope
Microscopes with an inverted-style frame are designed primarily for tissue culture applications and are capable of producing fluorescence illumination through an episcopic and optical pathway.
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Reflected Light Fluorescence Microscopy Light Pathways
In reflected light Köhler illumination, an image of the light source is focused by the lens onto the aperture iris diaphragm located in the vertical illuminator as seen in this tutorial.
Upright Epi-Fluorescence Microscope Light Pathways
Explore illumination pathways in an upright microscope. The microscope drawing presented in the tutorial illustrates a cut-away diagram of the Olympus upright microscope.
Inverted Epi-Fluorescence Microscope Light Pathways
Explore light pathways through an inverted tissue culture microscope equipped with for both diascopic (tungsten-halogen) and epi-fluorescence (mercury arc) illumination in this interactive tutorial
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