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dark field microscopy pdf

dark field microscopy pdf

dark field microscope

dark field microscopy pdf

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dark field microscopy

dark field microscopy pdf

What is dark field microscopy?

Similar to a bright field but it is modified by a dark field stop just below the source. The dark field stop is just the condenser, and blocks the light in the center of the lightsource so that the only light that goes through is around the edges. That light is then bent by the condenser and diffacts off the specimen. None of the light goes directly from the light source into the objective, so if there is no specimen, the image will be very dark. The specimen in this method will be illuminated against a black background.
A dark field microscope is useful because it increases the contrast of the image and does not use stains. The lack of staining means that it can be used on live specimens and that one can observe the motility of the organism as well as its correct morphology. Usually, the stains and enzymes used in labs can distort the shape of the organism, but that isn’t an issue with dark field microscopy. This method can also be used to see organisms that are hard to stain, such as Treponema pallidum, spirochetes, and mycoplasma.The one downside is that it’s not possible to see the inclusions, or internal details of the cell

What is Disadvantages of Dark Field Microscopy?

A dark field microscope can result in beautiful and amazing images; this technique also comes with a number of disadvantages.

First, dark field images are prone to degradation, distortion and inaccuracies.
A specimen that is not thin enough or its density differs across the slide, may appear to have artifacts throughout the image.
The preparation and quality of the slides can grossly affect the contrast and accuracy of a dark field image.
You need to take special care that the slide, stage, nose and light source are free from small particles such as dust, as these will appear as part of the image.
Similarly, if you need to use oil or water on the condenser and/or slide, it is almost impossible to avoid all air bubbles.
These liquid bubbles will cause images degradation, flare and distortion and even decrease the contrast and details of the specimen.
Dark field needs an intense amount of light to work. This, coupled with the fact that it relies exclusively on scattered light rays, can cause glare and distortion.
It is not a reliable tool to obtain accurate measurements of specimens.
Finally, numerous problems can arise when adapting and using a dark field microscope. The amount and intensity of light, the position, size and placement of the condenser and stop need to be correct to avoid any aberrations.

Dark field has many applications and is a wonderful observation tool, especially when used in conjunction with other techniques.

However, when employing this technique as part of a research study, you need to take into consideration the limitations and knowledge of possible unwanted artifacts.

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What is Advantages of Dark Field Microscopy?

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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What is Conclusion about dark field microscope

A dark field microscope can offer brilliant, light images against a dark background of otherwise difficult to view specimens.

Most standard microscopes come with dark field capabilities or accessories to enable this illumination technique.

There are many practical applications of dark field, especially in the field of marine biology, in viewing the many specimens you cannot see using alternative techniques.

However, a researcher must keep in mind the potential issues and limitations that may arise from dark field illumination.

For further information, check out the many microscopy imaging techniques available.

 

 

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