DIACHROMIC MICROSCOPE CONDENSER

CONDENSEUR POUR MICROSCOPE DIACHRONIQUE

Description

CA 02282865 1999-09-20
DIACHItOMIC MICROSCOPE CONDENSER
FIELD OF THE INVENTION
The present invention relates to light microscopy, and more particularly
concerns a
substage condenser allowing to illuminate in a certain fashion the very small
objects observed
(Appendix 1 ).
We recall, the specimen in light microscopy must be illuminated by a visible
light source.
Visibility of the objects (or particles) depends on three factors; the
object's absorption power, its
refraction and its diffraction.
TIZe absorption power defines the spectrum of light that is stopped by the
object, and can
be wavelength sensitive, which can influence the object's apparent color.
Refraction refers to the amount of light that is deviated by refraction from
its trajectory
when going through the object. if this refraction is weak, which is often the
case with very small
objects (or particles), the deviated light is too weak to be immediately
perceptible.
As for diffraction, it generates luminous circles around small objects which
become less
defined. With very small objects, the resulting image becomes a small and
diffuse spot which
does not become any clearer if the microscope's magnifying power is increased
(more and more
magnification gives a blur). In addition, when outside of normal range of the
resolving power of
the light microscopes, (appendix 2) the diffuse spot simply becomes invisible.
(Resolution power
being independent of the wavelength of the light source used, that is visible
light in this case, and
the numerical aperture of the objectives used).
Light sources in light microscopy generally consist of a beam of coherent
white light
projected from under the sample to be observed. Different substage condensers
offer various
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CA 02282865 1999-09-20
techniques of microscopy according to specific needs : (Example : bright field
illumination,
dark field illumination, phase contrast illumination, and differential
interference illumina-
tion). Neither of these previous techniques however allow to maximize the
visibility of the very
small sample (objects or particles).
SUMMARY OF THE INVENTION
The present invention therefore provides a substage condenser for any kind of
light
microscopes, a condenser that maximizes the visibility of the observed sample.
Specifically, this
is realized by generating a two-colored contrast in the light illuminating the
sample : this is the
bichromic contrast.
This new concept can be explained as follow : The incoming white visible light
source
(Figure 1- portion 22) is separated into two different beams.
THE FIRST BEAM : A first white beam (Figure 1 - portion 24) (wavelength X -
Ref.
Figure 4) deflected from its original trajectory to impinge on the sample from
its sides, and
achieved with aluminized surfaces.
A SECOND BEAM : A second straight forward beam travelling in a vertical light
path
(Figure 1- portion 26) (wavelength Y - Ref.: Figure 6) passing through a light
filter that lets only
a portion of the original spectrum through and impinging directly on the
sample, originating from
underneath.
SUMMARY : The concept behind this invention can be summarized by saying that
it is the result
of a certain design of the condenser's lens that permits the overlapping of
two different light
frequencies both emerging from one unique light source.
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BRIEF DESCRIPTION OF THE DRAWINGS
FIGURE 1 : Schematic view of the invention (piece 1-2-3 combined)
FIGURE 2 : Piece #1 (1 out of 3)
FIGURE 3 : Piece #2 (2 out of 3)
FIGURE 4 : Light spectrum of the acrylic material used for piece l and 2
FIGURE 5 : Piece #3
FIGURE 6 : Light spectrum of the Hesalite material used for piece 3
FIGURE 7 : Critical angles of refraction of piece # 1
FIGURE 8 : Schematic design and drawing of the universal support
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CA 02282865 1999-09-20
DESCRIPTION OF PREFERRED EMBODIIVVIENTS OF THE INVENTION
Referring to FIG. 1, there is shown the lens portion #IO of the condenser
(piece no 1,2,3
purposely assembled) according to a preferred embodiment of the invention. It
includes an
acrylic piece #12 (Figure 2) having its inside walls aluminized (14). Inside
the acrylic piece #12
is inserted no #16 (Figure 3), its wider end being inside the acrylic glass
piece #12.
Assembly of the three separate components of the lens (portion # 12, 16, 20)
creates an
inverted flare-shaped channel when the lens is in the upright position. The
outside walls of this
flare-shaped channel are also aluminized. At the widest end of the channel, we
find a light filter
(20), as described in figure 5. The light filter 20 is made of Hesalite H.T.
is transmissive only in
the higher portion of the spectrum of visible light. The other pieces #12, 16
(figure 2, figure 3)
are transmissive to the full spectrum of the visible light.
The present innovation is made possible by the contrast created by the
overlapping of
these two variations of transmissive qualities of the assembled pieces
designed according to the
preferred embodiment.
Said differently, the assembly of piece 12, 16 and 20 (len portion #10 of the
assembled
condenser) because of their respective design, when assembled according to the
referred
embodiment creates an inverted flare-shaped channel, thus permitting the light
path of the second
light beam to arrive directly on the light filter (20) to light up the
specimen at the focal point #28.
In use, the lens (10) is put into the trajectory of a beam of white light (22)
so that the
radius of said beam corresponds to the radius of the wider end of the flare-
shaped channel. In this
manner, an outer ring portion 24 of the beam 22 is reflected on the walls 18.
Whereas the center
portion 26 of the beam 22, enters the flare-shaped channel and goes in a
vertical path through the
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light filter 20. The critical angle of the outside walls 18 of the channel,
and the shape of the inside
walls 14 of the acrylic piece # 12 is calculated so that the outer ring
portion 24 of the beam
impinges on the outside walls 18 at the angle of maximum reflection, being
then also totally
reflected on the inside walls 14 of the acrylic piece #12 and finally focused
on a single point 28,
with which the sample should be aligned. The center portion 26 of the beam 22
following a direct
trajectory, it will also impinge on the point 28. The sample will therefore be
illuminated from
different angles by two beams of two different colors, originating from one
light source.
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Advantageously, for any light microscope, the Diachromic condenser according
to the
present invention, will provide a better contrast of the very small objects or
particles, and will
virtually eliminate the diffraction effects normally blurring the image.
In a second preferred embodiment, the actual light filter (20) is replaced by
another type
created with polarized material rather than the Hesalite (same design
different material). This
modification in the material used for the manufacturing process of the light
filter, when
used in combination with a commercial polarizer on the light source generates
a phase
difference between beams 24 and 26. By simply rotating the polarizer on the
light source, the
phase differences can easily be adjusted to any desired value. Producing the
second embodiment
has the advantage of having various types of illumination, without the need to
remove the
substage condenser. A specimen therefore can be observed in a specific field
permitting either
type of illumination (the darkfield, the phase contrast and the brightfield,
etc.) depending on one's
need. This feature is mostly requested in observation of live biological
material.
THE SUPPORT
The Diachromic condenser is called "universal" because it can be easily
adapted to all
existing brands of light microscopes. The design of the support is found at
Figure 8.
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HANUFA CT URER'S GUIDE AND TECHNICAL DA TA
Object : N~iessens " DIACHROMIC " Condenser
The assembled Naessens "Diachromic" Condenser, in its upright position.
Because the Naessens Condenser can be easily adapted to the major brands of
light microscopes, it is considered a "universal condenser".


CA 02282865 1999-09-20
MANUFACTURER'S GUIDE AND TECHNICAL DATA
Objeet : Naessens "Diachromic "Condenser
1. COMPONENTS : Optical and mechanical parts
~Ihe Nacsscns Uiachromic Condenser is composed of three different individual
parts
Lens holder with
thclens (na:l.4-1.2)
'the universal support
(with its tightening ring)
The adapter ring
(designed specifically for particular
brands of light mic~roscopcs)
2. OPTICAL CONCEPT : BICHROMICAL CONTRAST
'i'he Nacsscns Diachromic condenser is basui upon to the concept of bichronlic
contrast that
expresses in variations of luminance the overlapping of different luminous
frequencies emerging
from one light source. 'this condenser was specifically designed to enhance
the overall performance
of light microscopes used in the observation of live biological material. The
Naessens Diachromic
Condenser can also find many applications in the medical tlelds whenever a
diagnosis is required.
Although the Naessens Uiachromic condenser does not enhance the resolution
power of the light
microscope, it does make very small particles appear clearly as brilliant
objects on a magenta
background. (unique feature of the Naessens design). Morphological details arc
seen with extreme
sharpness, if the condenser is properly adjusted according to the
manufacturer's guidelines.
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CA 02282865 1999-09-20
3. APPLICATIONS
Other than the observation of live material, the Naessens Diachromic Condenser
can be used
~ In various fields of scientific research, mainly in biological sciences,
(ex: bacteriology, virology,
etc...)
~ In clinical microbiology and/or hematology
~ For industrial quality control
~ For all applications (medical or other) requiring the bichromic contrast
4. REQUIREMENTS FOR MAXIMUM PERFORMANCE
A) For optinrrrl perfornrarrce, the Naessens Diachromic Condenser needs to be
used with a light
microscope equipped with the following
i) A 100 Watt halogen lamp as the prime light source (not less)
ii) A 100X oil immersion objective (with an iris)
iii)A 4X scanning objective for the centering of the condenser
B) For optimal performance, the Naessens Diachromic Condenser must be used
with appropriate
microscopic material
i) Immersion oil
Tvpe : Low viscosity, low fluorescence immersion oil
Refractive index : nD = 1.516 at 23° Centigrade (if working with glass
specimen slides and
cover glass)
ii) Specirrren slide
Dimension (standards : 3 inches X 1 inch (75 millimeters X 25 millimeters)
Thickness : .035 inch to .040 inch (< 1 millimeter)
iii)Cover glass
Dimension : .700 inch X .700 inch (no bigger) (18 millimeters X 18
millimeters)
Thickness : .006 inch (0.152 millimeter) (Number 1 )
C) For optimal performance, when using the Naessens Diachromic Condenser, tire
doable oil
immersion teclrniyue is required. This technique can be summarized as follows
i) Prepare the specimen using the recommended specimen slide and cover glass.
ii) Once ready for microscopic observation, take the specimen slide in your
hand then turn the
specimen slide upside down and place a drop of immersion oil in the center
area of the slide
where the cover glass is located. When done, quickly turn the slide right side
up.
iii)Place a drop of immersion oil in the center area of the cover glass. Place
the specimen on the
stage.
Note : The oil on the condenser will make immediate contact with the oil on
the bottom part
of the slide. The oil on the upper part of the slide will make contact with
the 100X
objective when the stage is moved slightly upward. When all contacts occur
simultaneously, the specimen to be observed will be illuminated.


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5. ADJUSTMENT OF THE NAESSENS DIACHROMIC CONDENSER
STEP BY STEP INSTRUCTIONS
The adiustment of the condenser consists of three different steps
A) Inserting the condenser
B) binding the proper height of your condenser
C) Centering the condenser
A) To aroperly insert the condenser
Step 1 r~
Make sure that the adapter ring
provided with the condenser is compatible
with your brand of microscope.
(specify when ordering)
~ Step 2
:emove the condenser from its case
n box. Place the condenser in the
upright position with the lens facing
upward (the N.A. scale indicator
should face the front of the
microscope). 'rhe condenser is
ready to be inserted in its mounting
support.
~7 Step 4
Insert the condenser into the mounting
support just below the stage. It should
slide in easily. (If not, keep unscrewing
the adapter ring until the condenser slides
in without resistance.)
Step S ~
When fully inserted, tighten the adapter ring by turning it clockwise.
'I'hc condenser should be well inserted and the adapter ring tightly
secured in place.
4
Step 3
Slightly loosen the adapter
ring (turn counter-clockwise).


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'~7Step 6
Once the condenser is properly inserted (fully inserted on the horironlal
axis) you can lock the condenser in this position by using the clamping
screw usually located on the right side of the
condenser's support.
Leave the condenser at
the lowest possible
vertical position. 'I'hcn you can proceed to the next
adjustment.
13) To adiust tine height of'the condenser
'Ihe condenser needs to be perfectly located in terms of height in order to
create a perfect focal point
of the light path on the specimen, not below, nor above the specimen,
otherwise the image will be out
of focus (as for example, approximately 61 mm from bottom to top of condenser,
in the vertical
position for the Olympus microscopes).
IJvcn though the condenser has been adjusted in terms of height (according to
the brand of
microscope) prior the packaging, it is recommended to verify this scuing
before using the condenser.
It may have been displaced during shipping procedures, making the height
shorter than reduested.
As a bench mark, the condenser when inserted, raised to its optimal position,
should make very light
contact with the slide placed on the stage.
Here arc the steps to follow in order to adjust properly the height of the
condenser.
Step 7 ~'
Place a clean slide on the stage.
Step 9 ~'
Step 8
Now, raise the condenser to its highest position. At that position,
the lens of the condenser should slightly touch the slide, just
enough to insert a thin paper between the slide and the stage.
If, at the highest position> the condenser does not touch the slide, your
condenser needs to be lengthened. Remove the condenser from the
suplxnt and gently unscrew the tightening ring at the base of the Lens tube;
and adjust accordingly (counter-clockwise). 'I'ighte,~n the condenser at the
selec;tcd height, again using the tightening ring (clockwise). Repeat
procedures from step 4 until the condenser slightly raises the slide at the
highest position. When done, you may proceed with the centering process.


CA 02282865 1999-09-20
C) To center the condenser
'Ihe final adjustment required is to make sure the light path reaching the
lens of the condenser is
centered properly. An uncentered condenser would bring a constant white shadow
on one side of the
slide material. It could impair the optimal optic performance expected. Here
is the prcxedure, step
by step, on how to center the condenser.
Preliminaries
~ 'Turn the microscope ON.
~ 'Turn the light to the maximum intensity.
~ Opcn the light diaphragm to the maximum position.
~ Select the 4X scanning objective on the microscope.
~ Make sure there is no slide on the stage and that the condenser is at the
lowest possible position.
Centering of the condenser
1. Look in the eyepieces making sure they are properly adjusted
to your eyesight.
2. Start raising lhc condenser slowly towards its highest position.
3. Check carefully, a magenta colored disk will appear in lhc
center area of a bright clear field. locus on that colored inner
circle.
4. Bring this inner circle as close as possible to
the perfect center position in the field of view.
'Ibis can be done with the two condenser centering knobs lcxated at the front
part of the
~~ condenser's support (right and left knobs). 'Ihcsc knobs are very sensitive
and require
. only slight adjustments.
5. Once the colored circle is perfectly centered, raise the condenser slowly
to its
upward position.
6. As the condenser progressively rises, the image of the colored circle will
take more
and more space, until it perfectly overlaps the full field of view. You can
still adjust the center
position of the colored circle (centering knobs), all.hough it is easier to do
when the circle is at its
smallest dimension.
The condenser is now perfectly centered and ready to use. If the microscope is
permanently stationed, the
condenser will remain centered. Regular adjusttnenl is needed if the
microscope is moved frequently.
Please note before rtsinQ the evndenser
Once these adjustments arc made, before using the microscope fvr specimen
observation, bring the
condenser at the maximum upright position and put a coating of immersion oil
on the Condenser's lens.
'This is necessary to perform the double oil immersion technique.
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CA 02282865 1999-09-20
6. CARE AND MAINTENANCE
Proper care of the I~'aessens Diachronic Condenser is recommended for optimal
observation of slide
material.
Periodically, the condenser needs to be cleaned thoroughly : either as a
regular maintenance program
or for storage in its wooden box.
To remove excessive oil on the lens surface, always use lens paper since this
material will not scratch
the lens (cotton tissue could leave fibers on the lens). To thoroughly remove
the oil (once or twice a
month) the lens paper can be soaked with a small amount of 94 % alcohol. Use
very sparingly.
Always wipe off the lens very carefully.
Never use xylene, ether, or acetone directly on the condenser : (same
restriction as for any optical
accessory). It could dissolve the mounting cements.
Never leave the condenser's lens (or any other optical accessory) directly
soaking in a cleaning
solution. "
When using the Naessens Diachronic Condenser on a daily basis, you don't need
to remove excess
oil every time you stop using the microscope. All that is needed is to place
an unused specimen slide
on the stage in order to keep a permanent contact with the condenser's lens.
This can be done by
putting a generous coating of immersion oil on the condenser's lens. This
technique will preserve the
life span of your condenser.
7. GUARANTEE
When operation and maintenance of the Naessens Diachronic Condenser are
performed according to
these guidelines, including the use of adequate microscopy material and proper
light source, the
Naessens Diachronic Condenser is fully guaranteed against any defect for a 3
year period calculated
from the date of purchase.
8. MANUFACTURER'S DISCLAIMER
The Naessens Diachronic Condenser is an optical accessory. Owning such a
condenser does not
automatically confer the required skills to perform reliable live blood
testing.
CERBE inc., manufacturer of the Naessens Diachronic Condenser declines any and
all
responsibilities associated with blood interpretation or any diagnosis
performed with the use of the
condenser.
9. MANUFACTURER'S RECOMMENDATIONS
It is recommended that you carefully read the instruction manual of your
microscope.
To explore the numerous possibilities of this condenser, it is recommended to
be familiar with the
basic notions of microscopy*. (see Appendix t) including minimal experience
handling light microscopes.
7