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dark field microscopy blood what can analysis?

What is dark field microscopy?

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy 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.dark field microscopy 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).

 

How dark field microscopy work?

Microscopes are used to magnify objects. Through magnification, an image is made to appear larger than the original object. The magnification of an object can be calculated roughly by multiplying the magnification of the objective lens times the magnification of the ocular lens. Objects are magnified to be able to see small details. There is no limit to the magnification that can be achieved; however, there is a magnification beyond which detail does not become clearer. The result is called empty magnification when objects are made bigger but their details do not become clearer. Therefore, not only magnification but resolution is important to the quality of the information in an image.

The resolving power of the microscope is defined as the ability to distinguish two points apart from each other. The resolution of a microscope is dependent on a number of factors in its construction. There is also an inherent theoretical limit to resolution imposed by the wavelength of visible light (400-600nm). The theoretical limit of resolution (the smallest distance able to be seen between two points) is calculated as:

Resolution = 0.61 l/N.A.

where l represents the wavelength of light used and N.A.is the numerical aperture. The student-grade microscopes generally have much lower resolution than the theoretical limit because of lower quality lenses and illumination systems.

Standard brightfield microscopy relies upon light from the lamp source being gathered by the substage condenser and shaped into a cone whose apex is focused at the plane of the specimen. Specimens are seen because of their ability to change the speed and the path of the light passing through them. This ability is dependent upon the refractive index and the opacity of the specimen. To see a specimen in a brightfield microscope, the light rays passing through it must be changed sufficiently to be able to interfere with each other which produces contrast (differences in light intensities) and, thereby, build an image. If the specimen has a refractive index too similar to the surrounding medium between the microscope stage and the objective lens, it will not be seen. To visualize biological materials well, the materials must have this inherent contrast caused by the proper refractive indices or be artificially stained. These limitations require instructors to find naturally high contrast materials or to enhance contrast by staining them which often requires killing them. Adequately visualizing transparent living materials or thin unstained specimens is not possible with a brightfield microscope.

dark field microscopy relies on a different illumination system. Rather than illuminating the sample with a filled cone of light, the condenser is designed to form a hollow cone of light. The light at the apex of the cone is focused at the plane of the specimen; as this light moves past the specimen plane it spreads again into a hollow cone. The objective lens sits in the dark hollow of this cone; although the light travels around and past the objective lens, no rays enter it (Fig. 1). The entire field appears dark when there is no sample on the microscope stage; thus the name dark field microscopy. When a sample is on the stage, the light at the apex of the cone strikes it. The image is made only by those rays scattered by the sample and captured in the objective lens (note the rays scattered by the specimen in Figure 1). The image appears bright against the dark background. This situation can be compared to the glittery appearance of dust particles in a dark room illuminated by strong shafts of light coming in through a side window. The dust particles are very small, but are easily seen when they scatter the light rays. This is the working principle of dark field microscopy and explains how the image of low contrast material is created: an object will be seen against a dark background if it scatters light which is captured with the proper device such as an objective lens.

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

The highest quality darkfield microscopes are equipped with specialized costly condensers constructed only for darkfield application. This darkfield effect can be achieved in a brightfield microscope, however, by the addition of a simple “stop”. The stop is a piece of opaque material placed below the substage condenser; it blocks out the center of the beam of light coming from the base of the microscope and forms the hollow cone of light needed for darkfield illumination.

 

dark field microscopy Options Accessories

 

Metallurigcal reflected light brightfield/dark field microscopy.
Metallurgical reflected and transmitted light brightfield/dark field microscopy.
Stereo microscope 420 with darkfield attachment.
Stereo Zoom SMZ-168 microscope with darkfield attachment.
Biological laboratory phase contrast microscope with darkfield for up to 40x.
Biological laboratory microscope BA210 with darkfield slider.
Biological student microscope 162 with darkfield attachment.
Already have a microscope, but your microscope manufacturer does not make a darkfield stop? If there is a filter holder below your condenser, a darkfield stop we carry may work. Or you can mount a coin or circle of another opaque material in the center of a clear disk and put it in the filter holder.

 

dark field microscopy APPLICATIONS

 

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

 

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

What is dark field microscopy Blood Analysis?

You may find it difficult to locate many medical doctors that use this technique. The FDA does not approve of dark field microscopy blood analysis, therefore many doctor’s hands are tied. Viewing a fresh, natural blood sample (a sample not altered with any stains, etc., needed for normal microscopic exams), under the technology of a dark field microscope, will reveal conditions of your blood not normally even considered during the diagnosis of a normal blood test performed in doctor’s office or a lab.

However, an increasing number of health professionals have found that the use of this technique allows inspection of cellular dynamics which as noted above normally escape analysis or diagnosis using orthodox medical tests.

A dark field microscope is a microscope designed to permit diversion of light rays and illumination, from the side, so that details appear light against a dark background; as opposed to light passing straight through the specimen. If bright lights from the microscope pass directly through the specimen, the heat from the light source will kill the red blood cells (RBC)s faster. Also, by diverting the light rays, a greater amount of depth and details can be viewed. (Almost like a three-dimension view).

 

Dark Field Microscopy thus allows a health professional to evaluate the shapes and other properties of individual blood cells, indicating nutritional conditions which can be adversely affecting a person’s health. The advantage of this analysis over standard blood tests, which detect chemical changes in the blood, is the ability of dark field microscopy to detect nutritional disorders sooner, when the problem is in its infancy stages. By monitoring the blood’s condition, a health professional can assist in “balancing” the blood by giving dietary and lifestyle recommendations which can enhance health.

 

This microscopic photograph of healthy, powerful blood shows the red blood cells to be round, evenly shaped and freely floating in plasma. The plasma itself is clear with a few fat globules. There are no signs of clotting, bacteria, fungus, disease or stress. This is the kind of blood a healthy person should have flowing through their circulatory system

 

In darkfield microscopy, one is therefore able to observe “live blood.” Unlike the techniques of electron microscopy, no fixative is used so the picture is one of mobility rather than fixity. With stains and fixatives, the picture reveals a moment in time rather than a continuum.

What one sees in the mobile situation are the usual red blood cells, white blood cells, plasma—and what is floating in the plasma. Microbial activity, undigested food, fungi, and crystals are all apparent as is the capacity of the red blood cells to circulate and the white blood cells to devour morbid matter.

 

As we know, red blood cells transport oxygen to the tissues of the body. Without oxygen, we are devitalized, and according to some theories, the tissues go into a morbid state in which they can survive on fermentation rather than oxygenation. This is what is referred to as anaerobic and it is believed, by such persons as Nobel laureate Prof. Otto Warburg, that cancer thrives in such oxygen deficient conditions.

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

dark field microscopy blood

With darkfield microscopy, one often sees sees a condition called “rouleau” in which the red blood cells are stacked together as shown below. Some people believe it is because of the stress on the body of poor metabolism and others believe it is due to this as well as pH (acid-alkaline balance), wrong dietary choices or the presence of excessively high levels of free radicals. In any event, it is usually correctable.

Another condition that is often revealed in these tests is one in which the activity of red blood cells is compromised because of infection, bacterial or viral. In some cases, the red blood cells are misshapen or debilitated by parasitic invasion.

In the photograph above, the “rouleau” effect shows that the red blood cells are clumped together and stacked like coins. Rouleau affects proper oxygenation because the red blood cells do not circulate well enough to deliver oxygen where it is needed.

The condition also favors the growth of unhealthy organisms that can survive in a milieu that is less oxygen rich. Fungi, bacteria, and viruses require less oxygen than healthy tissue.

In the case of rouleau, since oxygenation is really critical to well being, the right diet and herbs may alleviate one of the underlying factors that contributes to cancer. However, enzymes, avoidance of the wrong foods, and protocols that address the specific issues of the patient would be expected to be more effective than more random efforts to ward off ill health.

For instance, one may or may not be iron deficient, but one may have room for improvement in diet and digestion as well as perhaps liver and immune function. Detoxification and decongestion can also be helpful.

 

Typically, a detoxifying herb will also be decongesting and sometimes also somewhat anti-parasitic, but not all herbal alkaloids are the same and not all formulae have the same actions. Therefore consultation with a practitioner who is knowledgeable in the areas that are pertinent is practical and, more importantly, often wise!

 

If the real problem is infection—and devitalization or cancer are secondary to infection—it is important to address the infection so that the red blood cells can “get back to their primary task,” which, of course, is to deliver oxygen to the tissues.

 

The idea that cancer is a disease of degeneration has had its fashionable phases and its days of rejection. The issue of whether an abnormal condition could perpetuate itself in a healthy internal environment, what is called “biological terrain” in the literature, is also debated but not resolved.

 

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.

dark field microscopy The Major Manufacturers

The major microscope manufacturers all have devices capable of dark field illumination. Depending on the make/model, the microscope may come with attachments or have the options for dark field accessories. The major companies are:

MAIKONG
MONKON
Nikon
Olympus
Ziess
Leica
Meiji

In addition, lesser known smaller companies that produce dark field microscopes are Quanfa Scientific Instrument Company, Proway Optics & Electronics and Dewinter Optical Inc.

The leading innovators in microscopes are Nikon and Olympus, who both offer stereo and compound microscopes with dark field capability and/or accessories.

 

 

How to Make a dark field microscopy?

You don’t need to buy a huge expensive set-up to experiment with dark field illumination.

To create a dark field, an opaque circle called a patchstop is placed in the condenser of the microscope. The patchstop prevents direct light from reaching the objective lens, and the only light that does reach the lens is reflected or refracted by the specimen. Easy enough, right?

If you want to make a dark field microscopy you’ll first need a regular light microscope. Below is your full list of “ingredients”:

dark field microscopy Microscope
Hole punch
Black construction paper
Transparency film
Glue
Scissors
Pen

Now use the following steps to make your patchstop:

Set up your microscope and choose the lowest-power objective lens.

Set the eyepiece aside somewhere safe.

Open the diaphragm as wide as possible. Then slowly close it until is just encroaches on the circle of visible light.

Now bend over and take a look at the diaphragm from below. See that opening? It’s only slightly smaller than the finished patchstop you’ll create.

Punch a few circles in the black construction paper with the hole punch. Measure one against the diaphragm opening. If it’s more than 10% larger, cut it down to about that size (10% larger than the diaphragm opening). If it’s smaller, cut out a larger circle.

Cut a 5 cm square of transparency paper.

Glue the black circle onto the transparency film, about 2 cm from the corner of the square. In that free 2 cm of paper, write the correct magnification power of your objective.

Mark the patchstop with the correct magnification power.

Repeat the above steps for all the objective powers except the oil immersion lenses.

Now use your patchstop to turn a light field unit into a dark field microscopy:

Select the correct patchstop for the objective power to be used.

Slip the patchstop between the filter holder and condenser. If your microscope has no filter, hold it manually below the condenser.

Remove the eyepiece.

Open the diaphragm and move the patchstop until the light is blocked entirely. Use tape to secure it if there is no condenser on your microscope.

Replace the eyepiece and examine the sample.

As you can see, a dark field microscopy can let users see specimens in a whole new way, bringing those into focus that don’t stand out under intense light. Using dark field illumination can open up a whole new view of microscopy.

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