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Specialized Microscopy Techniques

Section Overview:

Modern microscopists have developed a wide spectrum of useful techniques designed to aid in contrast enhancement and provide better observation and photomicrography of specimens. This section of the Microscopy Primer describes many of these techniques in detail.

Contrast Enhancing Techniques

  • Contrast in Optical Microscopy

    With the assistance of Dr. Robert Hoffman, we review the problems of contrast enhancement with both amplitude and phase specimens and review techniques that have been developed to assist with specimen contrast.

  • Darkfield Microscopy

    The following section discusses various aspects of the theory and practice of condenser design and other important concepts in both transmitted and reflected light darkfield microscopy.

  • Differential Interference Contrast

    A mechanism for rendering contrast in transparent specimens, differential interference contrast (DIC) microscopy is a beam-shearing interference system in which the reference beam is sheared by a tiny amount.

  • Hoffman Modulation Contrast Microscopy

    Learn more about Hoffman Modulation Contrast which was invented by Dr. Robert Hoffman. This contrast-enhancing technique serves to increase specimen visibility and contrast, especially for unstained and living specimens.

  • Oblique or Anaxial Illumination

    Achieving conditions for oblique illumination, which has been employed to enhance specimen visibility since the dawn of microscopy, can be accomplished by a variety of techniques with a simple transmitted optical microscope.

  • Phase Contrast Microscopy

    Phase contrast provides an excellent method of improving contrast in unstained biological specimens without significant loss in resolution, and is widely utilized to examine dynamic events in living cells.

  • Polarized Light Microscopy

    Polarized Light Microscopy exploits optical properties of anisotropy to reveal detailed information about the structure and composition of materials, which are invaluable for identification and diagnostic purposes.

  • Rheinberg Illumination

    First described around the turn of the century by British microscopist Julius Rheinberg, this technique provides beautiful high-contrast colored images of unstained specimens.

  • Introduction to Confocal Microscopy

    Confocal microscopy offers the ability to control depth of field, elimination or reduction of background information away from the focal plane, and the capability to collect serial optical sections from thick specimens.

  • Applications in Confocal Microscopy

    The range of applications available to Laser Scanning Confocal Microscopy include a variety of studies in neuroanatomy, neurophysiology, as well as morphological studies of a wide spectrum of cells and tissues.

  • Near-Field Scanning Optical Microscopy

    For ultra-high optical resolution, near-field scanning optical microscopy (NSOM) is currently the photonic instrument of choice. Near-field imaging occurs when a sub-micron optical probe is positioned a very short distance from the sample and light is transmitted through a small aperture at the tip of this probe.

Fluorescence Microscopy Techniques

  • Fluorescence Microscopy

    Fluorescence illumination is the most rapidly expanding microscopy technique employed today, both in the medical and biological sciences, a fact which has spurred the development of more sophisticated microscopes.

  • Fluorescence & DIC Combination Microscopy

    Fluorescence microscopy can be combined with contrast enhancing techniques such as DIC illumination to minimize the effects of photobleaching by locating a specific area of interest in a specimen using DIC.

  • Fluorescence & Phase Contrast Combination Microscopy

    Fluorescence microscopy can be combined with phase contrast to minimize the effects of photobleaching by locating a specific area of interest in a specimen using phase contrast.

  • Specialized Microscopy Techniques Interactive Tutorials

    Discover and explore this gallery of featured interactive java tutorials/applets designed to aid students in understanding difficult concepts in various specialized microscopy techniques.

Contributing Authors

Mortimer Abramowitz - Olympus America, Inc., Two Corporate Center Drive., Melville, New York, 11747.

Robert Hoffman - Modulation Optics, Inc., 100 Forest Drive, Greenvale, New York 11548.

Kenneth R. Spring - Scientific Consultant, Lusby, Maryland, 20657.

Kirill I. Tchourioukanov and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.

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