From the
Annual Report of the Board of Regents of The Smithsonian
Institution - 1944
The
Universal Microscope
It is only a reasonable supposition,
but already, in one instance, a very successful and highly
commendable achievement on the part of Dr. Royal Raymond
Rife of San Diego, California, who, for many years, has
built and worked with light microscopes which far surpass
the theoretical limitations of the ordinary variety of
instrument, all the Rife scopes possessing superior ability
to attain high magnification with accompanying high
resolution.
The largest and most powerful of
these, the Universal Microscope, developed in 1933, consists
of 5,682 parts and is so called because of its adaptability
in all fields of microscopical work, being fully equipped
with separate substage condenser units for transmitted and
monochromatic beam dark-field, polarized, and slit-ultra
illumination, including also a special device for
crystallography. The entire optical system of lenses and
prisms as well as the illuminating units are made of
block-crystal quartz, quartz being especially transparent to
ultraviolet radiations.
This illuminating unit used for
examining the filterable forms of disease organisms contains
14 lenses and prisms, 3 of which are in the high-intensity
incandescent lamp, 4 in the Risley prism, and 7 in the
achromatic condenser which, incidentally, has a numerical
aperture of 1.40. Between the source of light and the
specimen are subtended two circular, wedge-shaped,
block-crystal quartz prisms for the purpose of polarizing
the light passing through the specimen, polarization being
the practical application of the theory that light waves
vibrate in all planes perpendicular to the direction in
which they are propagated.
Therefore, when light comes into
contact with a polarizing prism, it is divided or split into
two beams, one of which is refracted to such an extent that
it is reflected to the side of the prism without, of course,
passing through the prism while the second ray, bent
considerably less, is thus enabled to pass through the prism
to illuminate the specimen.
When the quartz prisms on the
universal microscope, which may be rotated with vernier
control through 360 degrees, are rotated in opposite
directions, they serve to bend the transmitted beams of
light at variable angles of incidence while, at the same
time, a spectrum is projected up into the axis of the
microscope, or rather a small portion of the spectrum to the
other, going all the way from the infrared to the
ultraviolet.
NOW, WHEN THAT PORTION OF THE SPECTRUM
IS REACHED IN WHICH BOTH THE ORGANISM AND THE COLOR BAND
VIBRATE IN EXACT ACCORD, ONE WITH THE OTHER, A DEFINITE
CHARACTERISTIC SPECTRUM IS EMITTED BY THE ORGANISM.
In the case of the filter-passing form
of the BACILLUS TYPHOSUS, for instance, A BLUE SPECTRUM IS
EMITTED AND THE PLANE OF POLARIZATION DEVIATED PLUS (+) 4.8
DEGREES.
The predominating chemical
constituents of the organism are next ascertained after
which the quartz prisms are adjusted or set, by means of
vernier control, to minus (-) 4.8 degrees (again in the case
of the filter-passing form of the BACILLUS TYPHOSUS) so that
the opposite angle of refraction may be obtained.
A MONOCHROMATIC BEAM OF LIGHT,
CORRESPONDING **EXACTLY** TO THE FREQUENCY OF THE ORGANISM
(for Dr.Rife has found that EACH DISEASE ORGANISM RESPONDS
TO AND HAS A DEFINITE AND DISTINCT WAVE LENGTH, a fact
confirmed by British medical research workers) IS THEN SENT
UP THROUGH THE SPECIMEN AND THE DIRECT TRANSMITTED LIGHT,
THUS ENABLING THE OBSERVER TO VIEW THE ORGANISM STAINED IN
ITS TRUE CHEMICAL COLOR and revealing ITS OWN INDIVIDUAL
STRUCTURE IN A FIELD WHICH IS BRILLIANT WITH LIGHT.
The objectives used on the universal
microscope are a 1.12 dry lens, a 1.16 water immersion, a
1.18 oil immersion, and a 1.25 oil immersion. The rays of
light refracted by the specimen enter the objective and are
then carried up the tube IN PARALLEL RAYS through 21 light
bends to the ocular, A TOLERANCE OF LESS THAN ONE WAVE
LENGTH OF VISIBLE LIGHT ONLY BEING PERMITTED IN THE CORE
BEAM, OR CHIEF RAY, OF ILLUMINATION.
Now, instead of the light rays
starting up the tube in a parallel fashion, TENDING TO
CONVERGE AS THEY RISE HIGHER AND FINALLY CROSSING EACH
OTHER,arriving at the ocular SEPARATED BY CONSIDERABLE
DISTANCE as would be the case with an ordinary microscope,
IN THE UNIVERSAL TUBE THE RAYS ALSO START THEIR RISE
PARALLEL TO EACH OTHER BUT, JUST AS THEY ARE ABOUT TO PULL
THEM OUT PARALLEL AGAIN, ANOTHER PRISM BEING INSERTED EACH
TIME THE RAYS ARE ABOUT READY TO CROSS.
These prisms, inserted in the tube,
which are adjusted and held in alignment by micrometer
screws of 100 threads to the inch in special tracks made of
magnelium (magnelium having the closest coefficient of
expansion of any metal to quartz), are separated by a
distance OF ONLY 30 MILLIMETERS.
Thus, THE GREATEST DISTANCE THAT THE
IMAGE in the universal microscope IS PROJECTED THROUGH ANY
ONE MEDIA, EITHER QUARTZ OR AIR, IS 30 MILLIMETERS INSTEAD
OF THE 160, 180, OR 190 MILLIMETERS as in the empty or
air-filled tubes of an ordinary microscope, the total
distance which the light rays travel ZIGZAG FASHION through
the universal tube being 449 MILLIMETERS, although the
physical length of the tube itself is 229 millimeters.
It will be recalled that if one
pierces a black strip of paper or cardboard with the point
of a needle and then brings the card up close to the eye so
that the hole is in the optic axis, a small brilliantly
lighted object will appear LARGER AND CLEARER, REVEALING
MORE FINE DETAIL, than if it were viewed from the same
distance without the assistance of the card.
This is explained by the fact that the
beam of light passing through the card is very narrow, the
rays entering the eye, therefore, being practically
parallel, whereas without the card the beam of light is much
wider and the DIFFUSION CIRCLES MUCH LARGER. It is this
principle of parallel rays in the universal microscope and
the resultant shortening of projection distance between any
two blocks or prisms plus the fact that objectives can thus
be substituted for oculars, these "oculars" being THREE
MATCHED PAIRS OF 10-MILLIMETER, 7-MILLIMETER, AND
4-MILLIMETER OBJECTIVES IN SHORT MOUNTS, which would make
possible not only the unusually high magnification and
resolution but which SERVE TO ELIMINATE ALL DISTORTION AS
WELL AS ALL CHROMATIC AND SPHERICAL ABERRATION.
Quartz slides with especially thin
quartz cover glasses are used when a tissue section or
culture slant is examined, the tissue section itself also
being very thin. An additional observational tube and ocular
which yield a magnification of 1,800 diameters are provided
so that that portion of the specimen which is desired to be
examined may be located so that the observer can adjust
himself more readily when viewing a section at a high
magnification.
The universal stage is a double
rotating stage graduated through 360 degrees in
quarter-minute arc divisions, the upper segment carrying the
mechanical stage having a movement of 40 degrees, plus or
minus. Heavily constructed joints and screw adjustments
maintain rigidity of the microscope which weighs 200 pounds
and stands 24 inches high, the bases of the scope being
nickel cast-steel plates, accurately surfaced, and equipped
with three leveling screws and two spirit levels set at
angles of 90 degrees. The coarse adjustment, a block thread
screw with 40 threads to the inch, slides in a 1 1/2
dovetail which gibes directly onto the pillar post.The
weight of the quadruple nosepiece and the objective system
is taken care of by the intermediate adjustment at the top
of the body tube. The stage, in conjunction with a hydraulic
lift, acts as a lever in operating the fine adjustment. A
6-gauge screw having 100 threads to the inch is worked
through a gland into a hollow, glycerine-filled post, the
glycerine being displaced and replaced at will as the screw
is turned clockwise or anticlockwise, allowing a 5-to-1
ratio on the lead screw. This, accordingly, assures complete
absence of drag and inertia. The fine adjustment being 700
times more sensitive then that of ordinary microscopes, the
length of time required to focus the universal ranges up to
1 1/2 hours which, while on first consideration, may seem a
disadvantage, is after all but a slight inconvenience when
compared with the many years of research and the hundreds of
thousands of dollars spent and being spent in an effort to
isolate and to look upon disease-causing organisms in their
true form.
Working together back in 1931 and
using one of the smaller Rife microscope having a
magnification and resolution of 17,000 diameters, Dr. Rife
and Dr. Arthur Isaac Kendall, of the department of
bacteriology of Northwestern University Medical School, were
able to observe and demonstrate the presence of the
filter-passing forms of BACILLUS TYPHOSUS. An agar slant
culture of the Rawlings strain of BACILLUS TYPHOSUS was
first prepared by Dr. Kendall and inoculated into 6 cc of
"Kendall" K Medium, a medium rich in protein but poor in
peptone and consisting of 100 mg.of dried hog intestine and
6 cc of tyrode solution (containing neither glucose nor
glycerine) which mixture is shaken well so as to moisten the
dried intestine powder and then sterilized in the autoclave,
15 pounds for 15 minutes, alterations of the medium being
frequently necessary depending upon the requirements for
different organisms. Now, after a period of 18 hours in this
K Medium, the culture was passed through a Berkefeld "N"
filter, a drop of the filtrate being added to another 6 cc.
of K Medium and incubated at 37 degrees C. Forty-eight hours
later this same process was repeated, the "N" filter again
being used, after which it was noted that the culture no
longer responded to peptone medium, growing now only in the
protein medium. When again, within 24 hours, the culture was
passed through a filter-the finest Berkefeld "W" filter, a
drop of the filtrate was once more added to 6 cc.of K Medium
and incubated at 37 degrees c., a period of 3 days elapsing
before a new culture was transferred to K Medium and yet
another 3 days before a new culture was prepared. Then,
viewed under an ordinary microscope, these cultures were
observed to be turbid and to reveal no bacilli whatsoever.
When viewed by means of dark-field illumination and
oil-immersion lens, however, the presence of small, actively
motile granules was established, although nothing at all of
their individual structure could be ascertained. Another
period of 4 days was allowed to elapse before these cultures
were transferred to K Medium and incubated at 37 degrees C
for 24 hours when they were then examined under the Rife
microscope where, as was mentioned earlier, the filterable
typhoid bacilli, emitting a blue spectrum, caused the plane
of polarization to be deviated plus 4.8 degrees. Then when
the opposite angle of refraction was obtained by means of
adjusting the polarizing prisms to minus 4.8 degrees and the
cultures illuminated by a monochromatic beam coordinated in
frequency with the chemical constituents of the typhoid
bacillus, small oval actively motile, bright turquoise-blue
bodies were observed at a magnificatinn of 5,000 diameters,
in high contrast to the colorless and motionless debris of
the medium. These observations were repeated eight times,
the complete absence of these bodies in uninoculated control
K Media also being noted.
To further confirm their findings,
Drs. Rife and Kendall next examined 18-hour-old cultures
which had been inoculated into K Medium and incubated at 37
degrees C., since it is just at this stage of growth in this
medium and at this temperature that the cultures become
filterable. And, just as had been anticipated, ordinary
dark-field examination revealed unchanged, long, actively
motile bacilli; bacilli having granules within their
substance; and free-swimming, actively motile granules;
while under the Rife microscope were demonstrated the same
long, unchanged, almost colorless bacilli; bacilli,
practically colorless, inside and at one end of which was a
turquoise-blue granule resembling the filterable forms of
the typhoid bacillus; and free-swimming, small, oval,
actively motile,turquoise-blue granules. By transplanting
the cultures of the filter-passing organisms or virus into a
broth, they were seen to change over again into their
original rodlike forms.
At the same time that these findings
of Drs. Rife and Kendall were confirmed by Dr. Edward C.
Rosenow, of the Mayo Foundation, the magnification with
accompanying resolution of 8,000 diameters of the Rife
microscope, operated by Dr. Rife, was checked against a
dark-field oil-immersion scope operated by Dr. Kendall and
an ordinary 2-mm. oil-immersion objective, x 10 ocular,
Zeiss scope operated by Dr.Rosenow at a magnification of 900
diameters. Examinations of gram and safranin-stained films
of culture of Bacillus typhosus, gram and safranin-stained
films of blood and of the sediment of the spinal fluid from
a case of acute poliomyelitis were made with the result that
bacilli, streptococci, erythrocytes, polymorphonuclear
leukocytes, and lymphocytes measuring nine times the
diameter of the same specimens observed under the Zeiss
scope at a magnification and resolution of 900 diameters,
were revealed with unusual clarity. Seen under the
dark-field microscope were moving bodies presumed to be the
filterable turquois-blue bodies of the typhoid bacillus
which, as Dr.Rosenow has declared in his report
(Observations on filter-passing forms of
Eberthella-typhi-Bacillus typhosus - and of the
streptococcus from poliomyelitis, Proc.Staff Meeting Mayo
Clinic, July 13, 1932), were so "unmistakably demonstrated"
with Rife microscope, while under the Zeiss scope stained
and hanging-drop preparations of clouded filtrate culture
were found to be uniformly negative. With the Rife
microscope also were demonstrated brownish-gray cocci and
diplococci in hanging-drop preparations of the filtrates of
streptococcus from poliomyelitis. These cocci and
diplococci, similar in size and shape to those seen in the
culture although of more uniform intensity, and
characteristic of the medium in which they had been
cultivated, were surrounded by a clear halo about twice the
width of that at the margins of the debris and of the
Bacillus typhosus.Stained films of filtrates and filtrate
sediments examined under the Zeiss microscope, and
hanging-drop, dark-field preparations revealed no organisms,
however. Brownish-gray cocci and diplococci of the exact
same size and density as those observed in the filtrates of
the streptococcus cultures were also revealed in
hanging-drop preparations of the virus of poliomyelitis
underthe Rife microscope, while no organisms at all could be
seen in either the stained films of filtrates and filtrate
sediments examined with the Zeiss scope or in hanging-drop
preparations examined by means of the dark-field. Again
using the Rife microscope at a magnification of 8,000
diameters, numerous nonmotile cocci and diplococci of a
bright-to-pale pink in color were seen in hanging-drop
preparations of filtrates of Herpes encephalitic virus.
Although these were observed to be comparatively smaller
then the cocci and diplococci of the streptococcus and
poliomyelitis viruses, they were shown to be of fairly even
density, size and form and surrounded by a halo. Again, both
the dark-field and Zeiss scopes failed to reveal any
organisms, and none of the three microscopes disclosed the
presence of such diplococci in hanging-drop preparation of
the filtrate of a normal rabbit brain. Dr. Rosenow has since
revealed these organisms with the ordinary microscope at a
magnification of 1,000 diameters by means of his special
staining method and with the electron microscope at a
magnification of 12,000 diameters. Dr. Rosenow has expressed
the opinion that the inability to see these and other
similarly revealed organisms is due, not necessarily to the
minuteness of the organisms, but rather to the fact that
they are of a nonstaining, hyaline structure. Results with
the Rife microscopes, he thinks, are due to the "ingenious
methods employed rather than to excessively high
magnification." He has declared also, in the report
mentioned previously, that "Examination under the Rife
microscope of specimens containing objects visible with the
ordinary microscope, leaves no doubt of the accurate
visualization of objects or particulate matter by direct
observation at the extremely high magnification obtained
with this instrument."
Exceedingly high powers of
magnification with accompanying high powers of resolution
may be realized with all of the Rife microscopes, one of
which, having magnification and resolution up to 18,000
diameters, is now being used at the British School of
Tropical Medicine in England. In a recent demonstration of
another of the smaller Rife scopes (May 16, 1942) before a
group of doctors including Dr. J.H. Renner, of Santa
Barbara, Calif.; Dr. Roger A. Schmidt, of San Francisco,
Calif.; Dr. Lois Bronson Slade, of Alameda, Calif.; Dr.
Lucile B. Larkin, of Bellingham, Wash.; Dr. E. F. Larkin, of
Bellingham, Wash.; and Dr. W. J. Gier, of San Diego, Calif.,
a Zeiss ruled grading was examined, first under an ordinary
commercial microscope equipped with a 1.8 high dry lens and
X 10 ocular, and then under the Rife microscope. Whereas 50
lines were revealed with the commercial instrument and
considerable aberration, both chromatic and spherical noted,
only 5 lines were seen with the Rife scope, these 5 lines
being so highly magnified that they occupied the entire
field, without any aberration whatsoever being apparent. Dr.
Renner, in a discussion of his observations, stated that
"The entire field to its very edges and across the center
had a uniform clearness that was not true on the
conventional instrument." Following the examination of the
grading, an ordinary unstained blood film was observed under
the same two microscopes. In this instance, 100 cells were
seen to spread throughout the field of the commercial
instrument while but 10 cells filled the field of the Rife
scope.
The universal microscope, of course,
is the most powerful Rife scope, possessing a resolution of
31,000 diameters and magnification of 60,000 diameters. With
this it is possible to view the interior of the 'pin-point'
cells, those cells situated between the normal tissue cells
and just visible under the ordinary microscope, and to
observe the smaller cells which compose the interior of
these pin-point cells. When one of these smaller cells in
magnified, still smaller cells are seen within its
structure. And when one of the still smaller cells, in its
turn, is magnified, it, too, is seen to be composed of
smaller cells. Each of the 16 times this process of
magnification and resolution can be repeated, it is
demonstrated that there are smaller cells within the smaller
cells, a fact which amply testifies as to the magnification
and resolving power obtainable with the universal
microscope.
More then 20,000 laboratory cultures
of carcinoma were grown and studied over a period of 7 years
by Dr. Rife and his assistants in what, at the time,
appeared to be a fruitless effort to isolate the
filter-passing form, or virus, which Dr. Rife believed to be
present in this condition. Then, in 1932, the reactions in
growth of bacterial cultures to light from the rare gasses
was observed, indicating a new approach to the problem.
Accordingly, blocks of tissue one-half centimeter square,
taken from an unulcerated breast carcinoma, were placed in a
circular glass loop filled with argon gas to a pressure of
14 millimeters, and a current of 5,000 volts applied for 24
hours, after which the tubes were placed in a 2-inch water
vacuum and incubated at 37 degrees C. for 24 hours. Using a
specially designed 1.12 dry lens, equal in amplitude of
magnification to the 2-mm. apochromatic oil-immersion lens,
the cultures were then examined under the universal
microscope, at a magnification of 10,000 diameters, where
very much animated, purplish-red, filterable forms,
measuring less then one-twentieth of a micron in dimension,
were observed. Carried through 14 transplants from K Medium
to K Medium, this B. X. virus remained constant; inoculated
into 426 Albino rats, tumors "with all the true pathology of
neoplastic tissue" were developed. Experiments conducted in
the Rife Laboratories have established the fact that these
characteristic diplococci are found in the blood monocytes
in 92 percent of all cases of neoplastic diseases. It has
also been demonstrated that the virus of cancer, like the
viruses of other diseases, can be easily changed from one
form to another by means of altering the media upon which it
is grown. With the first change in media, the B. X. virus
becomes considerably enlarged although its purplish-red
color remains unchanged.
Observation of the organism with an
ordinary microscope is made possible by a second alteration
of the media. A third change is undergone upon asparagus
base media where the B. X. virus is transformed from its
filterable state into cryptomyces pleomorphia fungi, these
fungi being identical morphologically both microscopically
to that of the orchid and of the mushroom. And yet a fourth
change may be said to take place when this cryptomyces
pleomorphia, permitted to stand as a stock culture for the
period of metastasis, becomes the well-known
mahogany-colored Bacillus coli.
It is Dr. Rife's belief that all
micro-organisms fall into 1 of not more then 10 individual
groups (Dr. Rosenow has stated that some of the viruses
belong to the group of the streptococcus), and that any
alteration of artificial media of slight metabolic variation
in tissues will induce an organism of one group to change
over into any other organism included in that same group, it
being possible, incidentally, to carry such changes in media
or tissues to the point where the organisms fail to respond
to standard laboratory methods of diagnosis. These changes
can be made to take place in as short a period of time as 48
hours. For instance, by altering the media - 4 parts per
million per volume - the pure culture of mahogany-colored
Bacillus coli becomes the turquoise-blue Bacillus typhosus.
Viruses of primordial cells of organisms which would
ordinarily require an 8-week incubation period to attain
their filterable state, have been shown to produce disease
within 3 days' time, proving Dr. Rife's contention that the
incubation period of a micro-organism is really only a cycle
of reversion.
He states:
"In reality, it is not the
bacteria themselves that produce the disease, but we
believe it is the chemical constituents of these
micro-organisms enacting upon the unbalanced cell
metabolism of the human body that in actuality produce
the disease. We also believe if the metabolism of the
human body is perfectly balanced or poised, it is
susceptible to no disease."
In other words, the human body itself
is chemical in nature, being comprised of many chemical
elements which provide the media upon which the wealth of
bacteria normally present in the human system feed. These
bacteria are able to reproduce. They, too, are composed of
chemicals. Therefore, if the media upon which they feed, in
this instance the chemicals or some portion of the chemicals
of the human body, become changed from the normal, it stands
to reason that these same bacteria, or at least certain
numbers of them, will also undergo a change chemically since
they are now feeding upon media which are not normal to
them, perhaps being supplied with too much or too little of
what they need to maintain a normal existence. They change,
passing usually through several stages of growth, emerging
finally as some entirely new entity - as different
morphologically as are the caterpillar and the butterfly (to
use an illustration given us). The majority of the viruses
have been definitely revealed as living organisms, foreign
organisms it is true, but which once were normal inhabitants
of the human body -living entities of a chemical nature of
composition.
Under the universal microscope disease
organisms such as those of tuberculosis, cancer, sarcoma,
streptococcus, typhoid, staphylococcus, leprosy, hoof and
mouth disease, and others may be observed to succumb when
exposed to certain lethal frequencies, coordinated with the
particular frequencies peculiar to each individual organism,
and directed upon them by rays covering a wide range of
waves. By means of a camera attachment and a motion-picture
camera not built into the instrument, many "still"
micrographs as well as hundreds of feet of motion-picture
film bear witness to the complete life cycles of numerous
organisms. It should be emphasized, perhaps, that invariably
the same organisms refract the same colors. when stained by
means of the monochromatic beam of illumination of the
universal microscope, regardless of the media upon which
they re grown. The virus of the Bacillus typhosus is always
a turquoise blue, the Bacillus coli always mahogany colored,
the Mycobacterium leprae always a ruby shade, the
filter-passing form of virus of tuberculosis always an
emerald green, the virus of cancer always a purplish red,
and so on. Thus, with the aid of this microscope, it is
possible to reveal the typhoid organism, for instance, in
the blood of a suspected typhoid patient 4 and 5 days before
a Widal is positive. When it is desired to observe the
flagella of the typhoid-organism, Hg salts are used as the
medium to see at a magnification of 10,000 diameters.
In the light of the amazing results
obtainable with this universal microscope and its smaller
brother scopes, there can be no doubt of the ability of
these instruments to actually reveal any and all
microorganisms according to their individual structure and
chemical constituents.
With the aid of its new eyes - the new
microscopes, all of which are continually being improved -
science has at last penetrated beyond the boundary of
accepted theory and into the world of the viruses with the
result that we can look forward to discovering new
treatments and methods of combating the deadly organisms -
for science dose not rest.
To Dr. Karl K. Darrow, Dr. John A.
Kolmer, Dr. William P. Lang, Dr. L. Marton, Dr. J. H.
Renner, Dr. Royal R. Rife, Dr. Edward C. Rosenow, Dr. Arthur
W. Yale, and Dr. V. K. Zworykin, we wish to express our
appreciation for the help and information so kindly given us
and to express our gratitude, also, for the interest shown
in this effort of bringing to the attention of more of the
medical profession the possibilities offered by the new
microscopes.
DISCUSSION:
OBSERVATIONS WITH THE RIFE MICROSCOPE
OF FILTER-PASSING FORMS OF MICROORGANISMS
by Edward C.
Rosenow, Rochester, Minnesota
Recently, I reported to the staff of
the Mayo Clinic the more important observation made during
three days, July 5, 6 and 7, 1932, spent in Dr.Kendall's
laboratory at Northwestern University Medical School,
Chicago. I went there at the invitation of Drs. Kendall and
Rife, to share with them their observations in a restudy of
the filter-passing forms of Eberthella typhi as seen with an
improved model of the Rife microscope. They asked me also to
bring with me my cultures of the streptococcus from
poliomyelitis.
I would like to repeat here that
portion of my report which had to do specifically with the
Rife microscope.
Owing to the novel and important
character of the work, each of us verified at every step the
results obtained. Microscopic examinations of suitable
specimens was made as a routine by Dr. Rife with his
high-power microscope, by Dr.Kendall with the oil immersion
dark field, and by myself with the ordinary Zeiss microscope
equipped with a 2 mm apochromatic oil immersion lens and 100
X 10 ocular giving a magnification of about 90 diameters.
Most observations with the Rife microscope were made at
8,000 diameters. In order to check the magnification, gram
and safranin stained films of cultures of Eberthella typhi,
of the streptococcus from poliomyelitis, and stained films
of blood, and of the sediment of the spinal fluid from a
case of acute poliomyelitis were examined. Bacilli,
streptococci, erythrocytes, polymorphonuclear leukoeytes and
lymphocytes were clearly seen, and in each instance were, as
nearly as could be examined with the 2 mm oil immersion at
about 900 diameters.
The following principles and methods
were stated by Dr. Rife as being essential in order to
visualize clearly the objects at this and higher
magnifications by direct observation. Spherical aberration
is reduced to the minimum and magnification greatly
increased by using objectives in place of oculars. Proper
visualization, especially of unstained objects, is obtained
by the use of an intense beam of monochromatic polarized
light created by rotating wedge-shaped quartz prisms placed
between the source of light and the substage quartz
condenser. Dispersion of the transmitted rays of light, as
they pass upward to the eye, is prevented by passing them
through a series of quartz erecting (90 degrees) prisms.
Projection of the rays of light through air is not greater
tan 30 mm at any point.
In my original report I summarized as
follows:
"There can be no question of
the existence of the filterable turquoise blue bodies of
Eberthella-typhi described by Kendall. They are not
visible by ordinary methods of illumination and
magnification, not because they are too small. but
rather, it appears, because of their peculiar
non-staining hyalin structure. Their visualization under
the Rife microscope is due to the ingenious methods
employed rather than to excessively high magnification.
Examination under the Rife microscope of specimens,
containing objects visible with the ordinary microscope,
leaves no doubt of the accurate visualization of objects
or particulate matter by direct observation at the
extremely high magnification (calculated to be 8,000
diameters) obtained with this instrument."
The findings under the Rife microscope
of cocci and diplococci in filtrated cultures of the
streptococcus from poliomyelitis, and in filtrates of the
streptococcus from poliomyelitis, and herpes encephalitis,
not detectable by the ordinary methods of examination, and
which resembled in form and size those found in the
respective cultures, and the absence of minute forms,
suggest that the filterable, inciting agent of these
diseases is not necessarily extremely small, as is
universally believed. Indeed, the filterable, inciting agent
may be the non-staining, highly plastic, hyaline stage of
the visible, stainable, cultivable organism, the
streptococcus.
It is, of course, possible that these
unstained, invisible forms revealed by ordinary methods of
examination are not the inciting agents or 'viruses' of
these diseases and that they represent merely the filterable
or other state of the streptococcus. A consideration of the
great difficulty one has in isolating the streptococcus and
demonstrating diplococci in lesions in these diseases and
the ease with which the bodies are found in the filtrate
indicate clearly that the `invisible' forms of the
streptococcus, if such they be, are present in large numbers
in the host, as in positive cultures of the streptococcus.
Their form, size and color are too characteristic and true
to type to permit considering the masarti factsor as being
expressive of etiologically unrelated, contaminating
streptococci. Non-infectivity of the filter-passing forms,
except in the cases of virus diseases, their presence in
large numbers in filtrates, both of cultures and of infected
tissues, and the great difficulty in obtaining the visible
forms in cultures of filtrates indicate that "invisible,"
filter-passing forms represent a certain stage in the
development of microorganisms.