Advantages and disadvantages

Dark field microscopy produces an image with a dark background.Dark field microscopy
is a very simple yet effective technique and well suited for uses involving live and
unstained biological samples, such as a smear from a tissue culture or individual water-
borne single-celled organisms. Considering the simplicity of the setup, the quality of
images obtained from this technique is impressive.The main limitation of dark field
microscopy is the low light levels seen in the final image. This means the sample must be
very strongly illuminated, which can cause damage to the sample.Dark field microscopy
techniques are almost entirely free of artifacts, due to the nature of the process. However
the interpretation of dark field images must be done with great care as common dark
features of bright field microscopy images may be invisible, and vice versa.While the
dark field image may first appear to be a negative of the bright field image, different
effects are visible in each. In bright field microscopy, features are visible where either a
shadow is cast on the surface by the incident light, or a part of the surface is less
reflective, possibly by the presence of pits or scratches. Raised features that are too
smooth to cast shadows will not appear in bright field images, but the light that reflects
off the sides of the feature will be visible in the dark field images.

Dark Field Viewing

Dark field optics are a low cost alternative to phase contrast optics. The contrast and
resolution obtained with inexpensive dark field equipment may be superior to what you
have with student grade phase contrast equipment. It is surprising that few manufacturers
and vendors promote the use of dark field optics.A dilute suspension of yeast cells makes
a good practice specimen for dark field optics, particularly when cultured with living
Paramecium.

Principle

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 (figure 2).
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 field as
opposed to bright field viewing.
You don't need sophisticated equipment to get a dark field effect, although the effect is
most dramatic when the occulting disk is built into the condenser itself. 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.

Dark Field Viewing

Dark field optics are a low cost alternative to phase contrast optics. The contrast and
resolution obtained with inexpensive dark field equipment may be superior to what you
have with student grade phase contrast equipment. It is surprising that few manufacturers
and vendors promote the use of dark field optics.

A dilute suspension of yeast cells makes a good practice specimen for dark field optics,
particularly when cultured with living Paramecium.

Principle

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 (figure 2).
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 field as
opposed to bright field viewing.
You don't need sophisticated equipment to get a dark field effect, although the effect is
most dramatic when the occulting disk is built into the condenser itself. 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.

Limitation of darkfield microscopy

The advantage of darkfield microscopy also becomes its disadvantage: not only the
specimen, but dust and other particles scatter the light and are easily observed. For
example, not only the cheek cells but the bacteria in saliva are evident in Figure 2D.
More care in sample preparation needs to be exercised in darkfield application. Glass
slides need to be thoroughly cleaned of extraneous dust and dirt. It may be necessary to
filter sample media (agar, water, saline) to exclude confusing contaminants. Sample
materials need to be spread thinly; too much material on the slide creates many
overlapping layers and edges making it difficult to interpret structures.To create the
image, this technique relies on scattered light from specimens. Color is lacking or
minimal; this can be disappointing to the viewer. The actual size of specimens is also
impacted; the width of objects becomes exaggerated

Darkfield Microscopy or Live Blood Analysis is a way of studying

live whole blood cells under a specially adapted microscope
that projects the dynamic image onto a video screen. This
allows you to view your inner terrain. Digestive, eliminative and
immune functions can be assessed as well as the presence of
bacteria and other micro-organisms.
The darkfield microscopic examination of the freshly taken live
blood is one of the most important examinations of the holistic
medicine applied at the Tara Centre. It enables us to view the
inner terrain (milieu) and to examine the functions of the red
blood cells. It also shows the evolutionary stages of the
smallest proteins (endobionts) which are found in every human
body. We are also able to see any developed structures such as
bacteria, virus and fungus. The darkfield examination shows
the state of the blood cells, endobionts and the plasma in a
functional and structural way, making bacterial processes and
fungal pre-stages in the blood clearly visible.

The darkfield examination is most suitable for the evaluation of

clients with chronic diseases; for children who are prone to
infections; for recurrent bacterial problems; for candida and
other fungal problems and also to answer questions concerning
chronic problems of toxicity (e.g. amalgam disturbances).

The darkfield microscope is a vital instrument for the

supervision of biological therapies. For example, therapy tests
can be made by adding blood directly to the medication and by
controlling its reaction. The examination is very motivating for
the client because the results can be discussed and
demonstrated live on the screen. It cannot be replaced by any
other blood examination, neither by normal microscope
examination, nor by blood tests sent to laboratories. The blood
rapidly changes its function with changes of the inner terrain
after taking a sample.