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What is dark field microscope?

What is dark field microscope?

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Dark Field Microscope is a microscope illumination technique used to observe unstained samples causing them to appear brightly lit against a dark, almost purely black, background.
When light hits an object, rays are scattered in all directions. The design of the dark field microscope is such that it removes the dispersed light so that only the scattered beams hit the sample.
The introduction of a condenser and/or stop below the stage ensures that these light rays will hit the specimen at different angles, rather than as a direct light source above/below the object.
The result is a “cone of light” where rays are diffracted, reflected and/or refracted off the object, ultimately, allowing you to view a specimen in dark field.
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.
Dark field can be used to study marine organisms such as algae and plankton, diatoms, insects, fibres, hairs, yeast, live bacterium, protozoa as well as cells and tissues and is ideal for live blood analysis enabling the practitioner to see much more than is possible with other lighting methods.

What is Advantages of 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 Microscope 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.

What Disadvantages of Dark Field Microscope

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.

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.

dark field microscopy live blood analysis

What Principles of Dark Field Microscope

To view a specimen in dark field, an opaque disc is placed underneath the condenser lens, so that only light that is scattered by objects on the slide can reach the eye. Instead of coming up through the specimen, the light is reflected by particles on the slide. Everything is visible regardless of color, usually bright white against a dark background.

Pigmented objects are often seen in “false colors,” that is, the reflected light is of a color different than the color of the object. Better resolution can be obtained using dark as opposed to bright field viewing.Sophisticated equipment is not necessary to get a dark field effect, but you do need a higher intensity light, since you are seeing only reflected light. At low magnification (up to 100x) any decent optical instrument can be set up so that light is reflected toward the viewer rather than passing through the object directly toward the viewer.

What PARTS of dark ground microscope?

These are three requisites for adopting an ordinary microscope for Dark ground illumination:
1 Dark ground condenser
2 Suitable light intensity lamp
3 Funnel stop (to reduce numerical aperture of objective)
1. To visualize Trepenoma pallidum in exudates.
2. To visualize Leptospira species in blood & urine.
3. To visualize Spirillum minus in blood.
4. To detect motility of other bacteria.

5. To identify sheathed microfilaria & other parasites

What you can expect from Dark Field Blood Analysis

Thus, the dark field blood analysis provides information about the function and structure of blood cells and plasma endobionts, as well as developing bacterial and fungal precursors. The dark field examination also indicates changes within the cell through hormonal and mineral deficiencies. It is particularly beneficial for the evaluation of patients with chronic diseases, and children with susceptibility to infection, or recurrent bacterial problems, such as Candida or other fungal diseases. Dark field blood analysis is crucial in answering questions related to any chronic, or toxic problems.

Dark field microscopy is also an important tool in biological therapies. It can be used to test the effect of certain medications, by adding the medication to a blood sample and analyzing the reaction that is produced. This investigation is extremely motivating for the patient, by allowing him to directly experience the diagnosis.

The effects of dark field microscopy can not be replaced by any other blood test, especially not by normal laboratory microscopic blood tests, sent in fixed samples, as the blood changes its function due to environmental changes, so the blood must be tested while it is still fresh. It is also important to evaluate the degeneration tendency of blood samples, especially when considering the behavior of tumors.

The presence of bacteria precursors, which are not disease-inducing, but increase the risk of future disease development, can also be found in the dark field examination. Therefore, this dark field examination of the blood is a valuable and necessary preventative measure.

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