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leptospira dark field microscopy

dark field microscopy is what?

What is dark field microscopy?-Have you ever heard of a dark field microscopy? While such a name may sound like a sci-fi gadget used to measure black holes, in reality it’s just a handy tool used to view certain types of translucent samples. The average microscope user may not know about the concept of dark field microscopy, yet it can shed new light on the old way of viewing specimens.
Most people who have survived a biology class know what a light field microscope is. This type of scope uses bright field illumination, meaning it floods the specimen with white light from the condenser without any interference. Thus the specimen shows up as a dark image on a light background (or white field if you will).

leptospira under dark field microscopy
leptospira dark field microscopy
dark field microscopy for leptospira
leptospira in dark field microscopy
dark field microscopy of leptospira

This type of unit works best with specimens that have natural color pigments. The samples need to be thick enough to absorb the incoming light; so staining is usually paired with this type of microscope.

Plankton illuminated with a dark field microscopyYet what if the specimen is light colored or translucent, like the plankton on the right? It certainly won’t stand out against a strong white background. Additionally, some specimens are just too thin. They cannot absorb any of the light that passes through them, so they appear invisible to the user. This is where the concept of dark field illumination comes in!

Rather than using direct light from the condenser, one uses an opaque disk to block the light into just a few scattered beams. Now the background is dark, and the sample reflects the light of the beams only. This results in a light colored specimen against a dark background (dark field), perfect for viewing clear or translucent details.

On a grand scale, the same thing happens every day when you look up at the sky. Do the stars disappear when it’s light out? Of course not! They’re still there, their brilliance blotted out by the mid-day sun.

If you’re still having a hard time visualizing this concept, think of a dusty room with the light on and the door open. You may feel the dust affecting your breathing, but you probably won’t see it flying through the air.

Now turn off the light and close the door to just a sliver, while leaving the light on in the adjacent room. If you look at that sliver of light coming through the door, you’ll see all sorts of dust motes suspended in it. You’re employing a similar principle when you use dark field illumination!

How to made the dark field microscopy ?

It is very easy to make dark field microscopy yourself. What you have to do is place an opaque round stop in the condenser. An easy way is to cut a piece of black paper and put it on a filter in your filterholder. You can put the stop on a piece of clear acetate sheet. You can even try to draw the stop on it with black paint. The most important thing is to have it big enough to stop all light going directly into the objective. Only the light that is reflected by the objects in the sample reaches the objective then. Stronger objectives are more difficult because their NA is often too high. The NA of your condenser should always be higher then the NA of the objective. If patch-stops of 8, 10, 12 and 15mm are made you can’t go wrong really. For objectives of around x10 the middle sizes prove best.If you like to make the patchstop as precise as possible: The best way is to set up as normal (brightfield), remove the eyepiece and close/open the substage iris until it is *just* visible. Then, either bending your neck over double, or carefully removing the condenser, measure the diameter of the iris diaphragm as it is now set. A pair of calipers is useful here. This diameter is that for the patch stop. Very often, to be on the safe side it is best to add about 10% to this figure, this avoids leakage, especially if you have no means of centering the stop in the filter holder. If you have a phase contrast condenser, the largest phase contrast annuli often make excellent patch stops for darkfield!The real connoisseurs must have recognized the skills of Klaus Kemp in the arranged (cleaned) diatom slide photographed by Mike Samworth.

 

leptospira dark field microscopy images

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

leptospira dark field microscopy

What Advantages and Disadvantages about dark field microscopy?

 

No one system is perfect, and dark field microscopy may or may not appeal to you depending on your needs.

Some advantages of using a dark field microscopy are:

Extremely simple to use
Inexpensive to set up (instructions on how to make your own dark field microscopy are below)
Very effective in showing the details of live and unstained samples
Some of the disadvantages are:

Limited colors (certain colors will appear, but they’re less accurate and most images will be just black and white)
Images can be difficult to interpret to those unfamiliar with dark field microscopy
Although surface details can be very apparent, the internal details of a specimen often don’t stand out as much with a dark field setup.

Below are contrasting examples of dark field (left) versus bright field (right) illumination of lens tissue paper. Note how they both create a different style of image.

Dark field illumination Bright field illumination

Admit it, by now you’re curious to check out your own dark field! You can create one with minimal time and effort. Just read on…

dark field microscopy where need it?

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

In darkfield microscopy, contrast is created by a bright specimen on a dark background. It is ideal for revealing outlines, edges, boundaries, and refractive index gradients but does not provide a great deal of information about internal structure. Ideal subjects include living, unstained cells (where darkfield illumination provides information not visible with other techniques), although fixed stains cells can also be imaged successfully. Darkfield imaging is particularly useful in haematology for the examination of fresh blood. Non-biological specimens include minerals, chemical crystals, colloidal particles, inclusions and porosity in glass, ceramics, and polymer thin sections.

What is Dark Field Microscopic Blood Analysis?

You may find it difficult to locate many medical doctors that use this technique. The FDA does not approve of dark field microscopic 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 microscopy, 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 microscopy 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.

 

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.

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