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The Truth About dark field microscopy images

dark field microscopy images

What is Blood and What Does it Do?

 

Two types of blood vessels carry blood throughout our bodies: The arteries carry oxygenated blood (blood that has received oxygen from the lungs) from the heart to the rest of the body.

The blood then travels through the veins back to the heart and lungs, where it receives more oxygen. As the heart beats, you can feel blood traveling through the body at your pulse points – like the neck and the wrist – where large, blood-filled arteries run close to the surface of the skin.

The blood that flows through this network of veins and arteries is called whole blood. Whole blood contains three types of blood cells:

Red Blood Cells
White Blood Cells
Platelets

These blood cells are mostly manufactured in the bone marrow (the soft tissue inside our bones), especially in the bone marrow of the vertebrae (the bones that make up the spine), ribs, pelvis, skull, and sternum (breastbone). These cells travel through the circulatory system suspended in a yellowish fluid called plasma (pronounced: plaz-muh). Plasma is 90% water and contains nutrients, proteins, hormones, and waste products. Whole blood is a mixture of blood cells and plasma.
Red Blood Cells

Red blood cells (RBCs, and also called erythrocytes, pronounced: ih-rith-ruh-sytes) are shaped like slightly indented, flattened disks. Red blood cells contain an iron-rich protein called hemoglobin (pronounced: hee-muh-glow-bun). Blood gets its bright red color when the hemoglobin in RBCs picks up oxygen in the lungs. As the blood travels through the body, the hemoglobin releases oxygen to the tissues. The body contains more RBCs than any other type of cell, and each has a life span of about 4 months. Each day, the body produces new RBCs to replace those that die or are lost from the body.

White Blood Cells

White blood cells (WBCs, and also called leukocytes, pronounced: loo-kuh-sytes) are a key part of the body’s system for defending itself against infection. They can move in and out of the bloodstream to reach affected tissues. The blood contains far fewer white blood cells than red cells, although the body can increase production of WBCs to fight infection. There are several types of white blood cells, and their life spans vary from a few days to months. New cells are constantly being formed in the bone marrow.

Several different parts of blood are involved in fighting infection. White blood cells called granulocytes (pronounced: gran-yuh-low-sytes) and lymphocytes (pronounced: lim-fuh-sytes) travel along the walls of blood vessels. They fight germs such as bacteria and viruses and may also attempt to destroy cells that have become infected or have changed into cancer cells.

Certain types of WBCs produce antibodies, special proteins that recognize foreign materials and help the body destroy or neutralize them. Someone with an infection will often have a higher white cell count than when he or she is well because more WBCs are being produced or are entering the bloodstream to battle the infection. After the body has been challenged by some infections, lymphocytes “remember” how to make the specific antibodies that will quickly attack the same germ if it enters the body again.
Platelets

Platelets (also called thrombocytes, pronounced: throm-buh-sytes) are tiny oval-shaped cells made in the bone marrow. They help in the clotting process. When a blood vessel breaks, platelets gather in the area and help seal off the leak. Platelets survive only about 9 days in the bloodstream and are constantly being replaced by new cells.

Drop of Blood

Blood also contains important proteins called clotting factors, which are critical to the clotting process. Although platelets alone can plug small blood vessel leaks and temporarily stop or slow bleeding, the action of clotting factors is needed to produce a strong, stable clot.

Platelets and clotting factors work together to form solid lumps to seal leaks, wounds, cuts, and scratches and to prevent bleeding inside and on the surfaces of our bodies. The process of clotting is like a puzzle with interlocking parts. When the last part is in place, the clot happens – but if only one piece is missing, the final pieces can’t come together.

When large blood vessels are severed (or cut), the body may not be able to repair itself through clotting alone. In these cases, dressings or stitches are used to help control bleeding.

In addition to the cells and clotting factors, blood contains other important substances, such as nutrients from the food that has been processed by the digestive system. Blood also carries hormones released by the endocrine glands and carries them to the body parts that need them.

dark field microscopy images

dark field microscopy images

RPR/VDRL/MHA-TP (Serologic Tests for Syphilis) Darkfield/FTA-ABS Microscopy

A variety of serologic tests for syphilis are available, including:

  • VDRL (Venereal Disease Research Laboratory)
  • RPR (Rapid Plasma Reagin)
  • FTA-ABS (Fluorescent Treponemal Antibody Absorption)
  • TP-MHA (Treponema Pallidum Microhemagglutination Assay)

Each differs the others in the precise substance being measured, complexity, and specificity. All are satisfactory for use in managing syphilis. Abnormals may be:

  • Reactive,
  • Weakly reactive, or
  • Bordeline

Whenever a screening test (RPR, VDRL) is positive, a more specific test (FTA-ABS, TP-MHA) should be used to confirm the test and rule out a “biologic false positive.”

A negative or “nonreactive” test may indicate:

  • The patient doesn’t have syphilis
  • The patient has syphilis, but is so early in the course of the disease that the test has not yet turned positive. In these cases, the test may never turn positive if the patient is effectively treated.
  • The patient had primary syphilis, had a positive test, was effectively treated, 6 months have passed and the test has now reverted back to negative.
  • The patient had secondary syphilis, had a positive test, was effectively treated, 12-18 months have passed and the test has now reverted back to negative.
  • The patient has syphilis, but his/her immune system is impaired.

A positive or “reactive” test may indicate:

  • The patient has syphilis.
  • The patient had syphilis, was effectively treated, but the test has not yet returned to negative:
    • With primarily syphilis, it typically takes about 6 months for the test to turn negative.
    • With secondary syphilis, it typically takes 12-18 months for the test to turn negative.
    • The longer syphilis remains untreated, the longer it will take for the test to return to normal, and the less likely it is to ever return to normal.
  • The patient has a biologic false positive (BFP)

dark field microscopy images

What is dark field microscopy images ?

 

dark field microscopy images 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).

dark field microscopy images

What is dark field microscopy images?

What is dark field microscopy images?

Bright-field microscopy is the simplest of all the optical microscopy illumination techniques. Sample illumination is transmitted (i.e., illuminated from below and observed from above) white light, and contrast in the sample is caused by attenuation of the transmitted light in dense areas of the sample. Bright-field microscopy is the simplest of a range of techniques used for illumination of samples in light microscopes, and its simplicity makes it a popular technique. The typical appearance of a bright-field microscopy image is a dark sample on a bright background, hence the name.

dark field microscopy images Light path

The light path of a bright-field microscope is extremely simple, no additional components are required beyond the normal light-microscope setup. The light path therefore consists of:

a transillumination light source, commonly a halogen lamp in the microscope stand;
a condenser lens, which focuses light from the light source onto the sample;
an objective lens, which collects light from the sample and magnifies the image;
oculars and/or a camera to view the sample image.

Bright-field microscopy may use critical or Köhler illumination to illuminate the sample.

dark field microscopy images Performance

Bright-field microscopy typically has low contrast with most biological samples, as few absorb light to a great extent. Staining is often required to increase contrast, which prevents use on live cells in many situations. Bright-field illumination is useful for samples that have an intrinsic color, for example chloroplasts in plant cells.Bright-field microscopy is a standard light-microscopy technique, and therefore magnification is limited by the resolving power possible with the wavelength of visible light.

dark field microscopy images Advantages

Simplicity of setup with only basic equipment required.
Living cells can be seen with bright-field microscopes

dark field microscopy images Limitations

Very low contrast of most biological samples.
The practical limit to magnification with a light microscope is around 1300X. Although higher magnifications are possible, it becomes increasingly difficult to maintain image clarity as the magnification increases.
Low apparent optical resolution due to the blur of out-of-focus material.
Samples that are naturally colorless and transparent cannot be seen well, e.g. many types of mammalian cells. These samples often have to be stained before viewing. Samples that do have their own color can be seen without preparation, e.g. the observation of cytoplasmic streaming in Chara cells.

dark field microscopy images Enhancements

Reducing or increasing the amount of the light source by the iris diaphragm.
Use of an oil-immersion objective lens and a special immersion oil placed on a glass cover over the specimen. Immersion oil has the same refraction as glass and improves the resolution of the observed specimen.
Use of sample-staining methods for use in microbiology, such as simple stains (methylene blue, safranin, crystal violet) and differential stains (negative stains, flagellar stains, endospore stains).
Use of a colored (usually blue) or polarizing filter on the light source to highlight features not visible under white light. The use of filters is especially useful with mineral samples.

dark field microscopy images

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