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dark field microscope uses Here’s a Quick Way to know

dark field microscope uses Here’s a Quick Way to know

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What is dark field microscope ?

What is a Dark Field Microscopy? The dark field microscopic examination of freshly collected, vital blood is a pillar of the Paracelsus Clinica al Ronc holistic medical diagnosis. It provides information on the internal milieu and function of the blood cells, as well as the amount and development of endobionts, from which microorganisms and more sophisticated structures, such as bacteria, fungi, and viruses, develop.Darkfield Illumination is a transmitted light technique that uses oblique light to illuminate the sample. Light that does not impinge on the sample is not collected by the objective and results in a dark background. Light that interacts with the sample is scattered (refracted, reflected, and/or diffracted) and is “bent” toward the objective collection angle. This light is collected by the objective and is seen as light spots or areas (resulting from scattered light) on a dark background. Contrast is therefore generated and the sample visualized.Darkfield illumination is provided to the sample by a specialized condenser. The simplest DF condenser has a Stop, or Annulus illuminating ring (A). Here, an opaque circle obscures the central portion of the condenser light path. This allows only light in a ring to illuminate the sample. The diameter of the central stop, and thus illuminating annulus, is such that the angle of light is greater than the collecting angle of the objective. Thus without a sample, no light is collected by the objective. This kind of DF stop is useful only for low magnification objectives (<20x).For higher magnification objectives, modifications of the Annular Stop are: B: Immersion paraboloid; C: immersion double mirror concentric; D: cardioid concentric. Gray cone represents the light reflected and refracted from the specimen and collected by the objective. Hatched areas represent glass. Light blocking stops (s) limit light transmission to a hollow cone. i: Immersion oil.; r: reflecting surfaces. (Ruzin,1999).

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why dark field microscope ?

A dark field microscope is ideal for viewing objects that are unstained, transparent and absorb little or no light.

These specimens often have similar refractive indices as their surroundings, making them hard to distinguish with other illumination techniques.

You can use dark field to study marine organisms such as algae and plankton, diatoms, insects, fibers, hairs, yeast and protozoa as well as some minerals and crystals, thin polymers and some ceramics.

You can also use dark field in the research of live bacterium, as well as mounted cells and tissues.

It is more useful in examining external details, such as outlines, edges, grain boundaries and surface defects than internal structure.

Dark field microscopy is often dismissed for more modern observation techniques such as phase contrast and DIC, which provide more accurate, higher contrasted images and can be used to observe a greater number of specimens.

Recently, dark field has regained some of its popularity when combined with other illumination techniques, such as fluorescence, which widens its possible employment in certain fields.

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dark field microscope uses?

Viewing blood cells (biological darkfield microscope, combined with phase contrast)
Viewing bacteria (biological darkfield microscope, often combined with phase contrast)
Viewing different types of algae (biological darkfield microscope)
Viewing hairline metal fractures (metallurgical darkfield microscope)
Viewing diamonds and other precious stones (gemological microscope or stereo darkfield microscope)
Viewing shrimp or other invertebrates (stereo darkfield microscope)

In darkfield microscopy, contrast is created by a bright specimen on a dark background. It is ideal for revealing outlines, edges, boundaries, and refractive index gradients but does not provide a great deal of information about internal structure. Ideal subjects include living, unstained cells (where darkfield illumination provides information not visible with other techniques), although fixed stains cells can also be imaged successfully. Darkfield imaging is particularly useful in haematology for the examination of fresh blood. Non-biological specimens include minerals, chemical crystals, colloidal particles, inclusions and porosity in glass, ceramics, and polymer thin sections.

Bright and Darkfield

 

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