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Everything I know about bright field dark field I learned from bright field dark field

Everything I know about bright field dark field I learned from bright field dark field

What is Dark Field microscope?

dark field microscopy of sugar crystals-Dark Field illumination is a technique used to observe unstained samples causing them to appear brightly lit against a dark, almost purely black, background.Pictured right: Highly magnified image of sugar crystals using darkfield microscopy technique,When light hits an object, rays are scattered in all azimuths or directions. The design of the dark field microscope is such that it removes the dispersed light, or zeroth order, 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.

What is Dark field illumination Applications ?

Viewing Blood cells.
Viewing bacteria.
Viewing different types of algae.
Viewing hairline metal fractures.
Viewing diamonds and other precious stones.
Viewing shrimp and other vertebrae.
Advantages and Disadvantages of Bright Field Microscopy.
Application of Bright Field Illumination
-This technique is widely used in pathology to
view fixed tissue sections or cell films/smears
-In biological applications, brightfield observation is widely used for stained or naturally pigmented or highly contrasted specimens mounted on a glass microscope slide.

Bright and Darkfield Bright and Darkfield

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.

When to use dark field illumination

Dark field illumination is most readily set up at low magnifications (up to 100x), although it can be used with any dry objective lens. Any time you wish to view everything in a liquid sample, debris and all, dark field is best. Even tiny dust particles are obvious. Dark field is especially useful for finding cells in suspension. Dark field makes it easy to obtain the correct focal plane at low magnification for small, low contrast specimens. Use dark field for

Initial examination of suspensions of cells such as yeast, bacteria, small protists, or cell and tissue fractions including cheek epithelial cells, chloroplasts, mitochondria, even blood cells (small diameter of pigmented cells makes it tricky to find them sometimes despite the color).
Initial survey and observation at low powers of pond water samples, hay or soil infusions, purchased protist or metazoan cultures.
Examination of lightly stained prepared slides. ? Initial location of any specimen of very small size for later viewing at higher power.
Determination of motility in cultures

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.

Difference between Dark and Bright Field Illumination

-When you view a particular specimen under a bright field microscope, you will observe that the specimen is dark while its background is bright; hence the name bright field microscope.
On the other hand, when you view a particular specimen under a dark field microscope, you will observe that the specimen is bright while its background is dark; hence the name dark field microscope.
-Since little light actually falls on the specimen, dark-field illumination shows less detail overall than bright-field illumination.

Bright Field Illumination.
A way of illuminating a specimen in a microscope by lighting it from behind, making the specimen appear dark against a bright background. It is considered the most basic type of microscope

The Dark field illumination requires blocking out the central light waves along the optical axis of the light waves. Blocking the light waves allows you to see the specimine when only the oblique rays hit the specimen at an angle.
Parts of the Microscope.
Monitor: To display the picture of the specimen your CMO objective lens is focused on.

CMO Objective lens: To magnify on the part of a specimen you wish to observe.

Stage: To hold your specimen.

Lamp Voltage: Controls how bright the light is.

Zoom Body: To zoom in on the specimen.

Illumination section: Illuminates the specimen.

Brightfield/Darkfield Diascopic Stand. Holds the inner pieces of the microscope inside.

Camera Control: Controls where the CMO Objective lens focuses on.

Economic observation tube: Allows you to see the specimen without the monitor.

Digital camera: Transfers the pixels onto the moniter so you can see the specimen.

Stage: Holds the microscope together and supports the microsope.

bright field dark field

difference between bright and dark field microscopy

bright field and dark field microscopy

 

 

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