Note: Descriptions are shown in the official language in which they were submitted.
` `~
sackground of the Invention
The invention pertains to a photometer microscope with
an apparatus for the microphotometer scanning of fine specimen-
structures, which, in the ray-path between the objective and
the photometer-part, exhibits an internal focusing lens
displaceable in a plane vertical to the optical axis.
Photolneter microscopes with an apparatus for the rnicro-
photometer scanning of small specimen-structures are used
for the study of diverse objects. One field of u-tilization,
for instance, is a study of chromosomes, inasmuch as for
some years now, methods e~ist with which it became possible
to produce on chromosornes transverse stripes or different
, color intensity. This coloration makes it possible to
. reliably distinguish the chromosome-pairs from each other,
and to correspondingly classify them.
¦ The differences in the color intensity of the transverse
stripes can be recognized with the naked eye only with gxeat
;` difficulty. For this reason photometer microscopes are
used, which can meas~re and graphically record the different
~7 ~ color intensities. For such an evaluation, it is required
that the object to be studied be moved for microphotometer
scanning, in order to be able to scan point by point the
interesting parts of the object specimen in corresponding
~` time sequence. The measured values are stored in a computer
during scanning at the conclusion of the scanning, the
information a~out the objects are compu-ter evaluated and
reported.
To produce the necessary relative movement, di~Eerent
devices have been used. In general, the procedure`is to
'
- 2 -
, . ~
~ I :
.
,~ ,
4~
secure the object on a motor driven stage. The motors can
be either direct curr~n~ motors which are controlled by a
joy-stick, or steppiny motors can be used. In tha first
case, one gets a con-tinuous, infinitely adjustable movement
of the motor stage, while in -the second case, the step
maynitude is not adjustable. with tasks with steppi~g
motors, a minimal s-tep size of 0.5 um is possible which is
not sufficiently small for the study of chromosomes, because
a step of 0.1 um is required. Consequently, work can only
be done with a movable stage, if realtively coarse objects
are to be studied.
; Despite the use of a movable stage, in order to make
I possible a finer scanning, it is also known to electronically
I increase the number of measurements during one step of the
stage. For example, if four measurements are -taken during
an 0.5 um step, effective distance is reduced to about 0.125
um. This procedure, can only be applied in one direction.
¦ Since the time available for a measurement by this system is
~aturally very short, evaluation of the results is difficult.
To overcome these problems the chromosomes or the like
are first photographed and then evaluated from the photos
.
thus obtained, which are larger than the originals, by the
photometer microscope. It is obvious that this procedure
also is not satisfactory, as it significantly increases time
and errors caa can creep in, for instance, throuyh defects
o~ the photographic emulsion.
- It is likewise known (CS-PS 129 178) how to provide in
the ray-path of the microscope ~etween the objective and the
photometer-part, an internal focusing lens, which moves the
image past the measuring diaphragm ~restrictor). This
:~ .
.' : .
t '
.. . . .. ,, .. . .:
' '
,' ~ ,
..
. . ' ' : ' ,, ,
.
h ~ ~
has the aclvantage that rel~tively sm~ll steps can b~
achieved, inasmuch as the already magniEied image is now
moved, hence the magnification factor of the objective can
remain neglected. A disadvantage of the known arrangement,
however is, above all, that through the additional lens, the
ray-path of the microscope becomes altered, so that the
microscope must be modified to accept the additional lens.
. Consequently, the known arrangemen-t cannot find acceptance
as an attachment for microscopes. Furthermore, it is
proposed to move the lens by hand using a pantograph-like
device. This has the substantial disadvantage that small,
uniform movement cannot be achieved although such movement
; i9 not required because measùrement takes place as a function
of the position of the pantograph. Naturally, the disadvantages
arises from this, that no continuous scanning of the specimen
, is possible, but only discrete areas, separated from each
: other, can be covered in any given case.
Finally, it is already known how to undertake an
imagç-shift by optical means in the image field, for which :
purpose glass wedges or prisms, or rotatable or:displaceable
deflecting mirrors, operating opposite each other, are
.: .
.~ provided. Here also, however, the problem arises that the
~ scanning steps, in general, do not become as fine as they
are required to be. Addi-tionally, devices having several
prisms or deflecting mirrors are of a projecting nature so
that only with difficulty if at all can they be brought into
the microscope tube, and above 211~ they are not suitable
- for utilization as an attachment.
.: . ' .
': j
.,'~ i
-- 4 --
. ~ :
` ,.
` ,'' :
~ !
, ` ~ ' -
: `
'
.
47
Summ~r~_oE the Invention
The present invention is based on a devlce for -the
micxophotometer scanning of fine specimen-structures, which
is constructed, so that the scanning can take place in fine
steps in either direction. In addition, the possibility of
a device constructed for the microphotometer scanning as an
attachment which can ~e used without difficulties on existing
microscopes.
According to the invention, a photometer microscope has
a stationary compe~sating lens in the ray-path (aY~is) directly
in front of or behind the in-tcrnal focusing lens of the same
focal pQwer but opposite that of the internal focusing lens.
With the photometer microscope according to the invention,
therefore, the displacement of the image occurs in front of
the photometer diaphragm over an internal focusing lens
inserted in series in the ray-path to the ob]ective, whereby,
despite relatively large displacement paths, an incremental
displacements can be achieved when compared to the prior
art. In spite of the presence of this internal focusiny
lens, however, the ray-path of the microscope in itselE is
not modified, hecause a compensatiny lens is provided. For
the first time~possible to equip a microscope with a scanning
system for the displace of the image in front of the photometer
diaphragm. Both lenses, that is, the movable internal
focusing lens and the stationary compensating lens, if they
are directly adjacent to each other can be inserted without
;~ difficulty in the ray-path, for instance, in an attachment
~ o an existing xic~oscope.
~ .. ~ "' .
~ ~ _ 5
. . .
. , ~ .
. . ~ .
,.
- ' ' , ." : ' ~' ', - ':
~ -' ' ' ' ' "' '
-' ' , :
.. ' .
: ~ :
~L .A3~ 7
~ ccording to the present invention there is provided a
scanning system ~or a microscope which compxise~, a pair of
` lenses on an optical axis each o~ the pair having equal and
opposite power~ one of thP pair ~eing movahle in a plane normal
to the optical axis.
':
:-.
''
.
. . ~
.
. . .
" -
.
,- .,
. P~
~ - 5a -
'' ' ' ` '
.: ,
. ~
Detaile~ Descri~tion_of the Invention
_
If the internal focusing lens and the compensating lens
are arranged in the ray-path between the objective and a
beam-splitter selectively positionable on the ray-path to
effect a division of the light for the photometer and ocular
which would have to be provided with a marking, for instance,
sS ~al~
a h~i~P~ or a graduated dial (plate), the possibllity
additionally exists o~ observing through the ocular the
displacement of the image in front of the photometer diaphragm,
so that it is possible to move the internal focusing lens
mutually or with a special device, in such a manner that the
movement of the specimen takes place along a preaetermined
track, for instance, a curved line.
A highly ùniEorm movement, which is very desirable for
computer-evaluation, can be obtained if the internal focusing
lens is movable by a motor in a plane normal to the optical
axis (ray-path). It is also advantageous for the drive to
have two motors which move the internal focusing lens in
perpendicular directions.
There are several types of motors which are appropriate
for this purpose. However, it is especially favorable if
the speed of rotation of both motors is continuously variable,
for instance, as with direct-current motors. The motors may
be controlled by a single control unit so that the movement
:
of the internal focusing lens is at a uniform speed in all
directions of movement. Then, the driving of the scanning
beam, for instance on a curved line, is facilitated.
A preferred control for the motors is a control stick
is advantageously used for both motors. If the control
stick is ball mounted in a recess, it can be used for regulating
'' . .
,,,, .
- 6 -
. ~ .
..,.
.. . .. ...
'~ '' ' ' . ' . . ~ :",, .
.: ~ . . ' . ' ' ,, : .' ,
' ~' ~ .- ' , .
- . ;,
- ~
the speed of both motors. Maximal deflection of the control
stick provides the highest speed oE movement and no deflection
of the stick gives no movement so that displacement of the
stick controls both the direction of movement as well as the
speed of movement of both motors.
In order to achieve this, a so-called "joy-stick" is
used, which can be constructed in such a way, that the
control-stick dispiaces two potentiometers or the like
positioned on perpendicular axes.
If a computer is used for the evaluation of the photometric
measured signals it can also be used for the control of the
motor drives for the interior Eocusing lens.
If, as is further provided according to the inven-tion,
the interior focusing lens and the compensa-ting lens are
arranged in an attachment which aan be laterally inserted in
:.
`~` a slot or some similar aperture of the microscope tube, the
` scanning device can be easlly disconnected if no further
movement is supposed to take place. Also, the slot or the
aperture can be empLoyed for the attachment of other attachments.
,, ~
. When the housing of the additional lenses is an attachment,
it is desirable that it contains the motors and other driving
.: :
-~ parts for movement of~the focusing lens, because the driving
`~ ~ - elements can always remain linked with the interior focusing
lens and thus, an ad~ustment once made can remain unchanged.
Finally, the fact lies within the sçope o~ the invention
. . , . ~
that the housing, which takes the motors and other driving
parts for the moving of the interior focusing lens, has on
`:~
the top a dlsplay for showing the position of the interior
focusing lens, for instance, to be able, without difficulties,
... i :
-~ to set the scanning beam before the start of the scanning,
in reference to the specimen.
''; ' ~ . .
.. , :
~, I :
1 7 ~ .
; :
.
.:
Fur~her fea-tures, details and advantages of the
invention are shown from the ~ollowing ~escription of a
preferred embodiment, reference being maae to the drawings.
The Drawings
Fig. 1 is a diagram of a photometer microscope
according to the invention;
Fig. 2 is a top view of a stained chromosome to be
studied;
Fig. 3 is a representative curve of the measured
photometric values;
Fia,. ~ shows an attachment having the interior focusing
lens, compensating lens, and their drive means; and
Fig. 5 shows a device for controlling the movement of
the interior focusing lens, and
Fig. 6 is a sketch illustratin~a, the operation of the
control de~lce of Flg. ~.
Detailed Descripti'on of *he Dr'awing
,~ The photometer microscope shown-'in Fig. 1 exhibits,
as sho~n, a microscope staa,e 1, on which the specimen 2 is held.
Along the ray-path ,(optical axis) 14 startina, from specimen 2 in
sequence, objective 3 of relatively high maynification, focusing
' lens 4, stationary lens 5, which is of the same focal power but
of opposite sign to lens 4. Interior focusina lens 4 is a
' convergina lens and compensatina lens 5 is a divergina, lens. ,
Following compensating lens 5, light-beam passes throuah optional
beam-splitter 6 to the ocular 7 and diverging lens 8, in ~ront of
mirror 9 having aperture 10 acting as a photometer diaphra~m.
Light passing through,the diaphragm 10 then reaches detector 11.
In the present case, a secondary-electron-multiplier is used, the
liaht is def~ec~ed by mirror 12 to detector.
bm:~,
Beam-splitter 6 is normaily located in the posi~ion
indicated by the dotted line in Fig. 1. If, however, no
visual observation is desired through ocular 7, it can be
moved into the position shown in the solid line, in which
case it is not located on optical axis 13. As is indicated
in Fig. 1, interior focusing lens 4 can be moved from the
position shown by the solid line, in which it is centered on
the optical axis 13, into tlle dotted line posi-tion 4a. The
consequence of this is, that for the scanning, the observation
axis 14 lying on optical axis 13 is not used, bu-t rather,
alternate observation axis 14a displaced a small distance in
the arrow-direction lS is used. Consequently, an area of
the specimen 2 is scanned, which is located to the right
from the image-point scanned by the beam 14. In conjunction
with movement of the intornal focusing lens 4 to alternate
location 4a, the sequence of images produced at measuring
diaphragm 10 and detector 11 is a sequence oE areas of
specimen 2, along the direction arrow 15.
The interior focusing lens 4 is moved by means of
.~,
motors ~not shown) in two directionsi one perpendicular to
the other, which directions lie in a plane, which is perpen-
dicular to the optical axis 13. The shifting of the observation
axis 14a for scanning in any direction afforded thereby
For example, chromosome 17 as shown in Fig. 2 may be scanned
along the curved path indicated by the arrow 18, so that
independent of the given curvature of the chromosome 17, a
scanning takes place over its entire length, without observation
beam 14a evex leaving the chromosome.
In conjunction with such scanning and recoxding of the
measured signals of photo multiplier 11 in a recorder, we
.' '; ' ' .
e _ g _
,
obtain on paper 19 curve 20 as shown in ~ig. 3. The curve
2Q gives in~ormation concerning thc differenti~l coloration
of chromosome 17 along the path 18. From curve 20, conclusions
can then be reached concerning the specific features of
specimen 2.
Another possibility is, naturally, to feed signals of
the photo multiplier 11 into a computer, which then expresses
corresponding values after the conclusion of the measuring
procedure, based on a given program. As was already
mentioned above, it is possible`in conjunction with the
photometer microscope in accordance with the invention, to
construct interior focusing lens 4 and compensating lens 5
tcgether with the driving elements for lens 4, as an attachment.
. , .
Fig. 4 shows the appearance of such an attachment. This
device comprises insert 21 which in the embodiment shown is
constructed in a rectangular shape and si~e chose to fit in
a recess in the microscope tube (body). Suh recesses
presently exist for receiving analyzers contract interference
devices, or the like. The insert 21 has two apertures 22
~one shown) in alignment between which lenses 4, 5 are
arranged. When;using attachment shown in Fig. 4 insert 21
is positioned so that apertures 22 and lenses 4, 5 are
located on the optical axis 13 of the microscope.
On distal end 23 o insert 21, a housing 24 is located
. .
to contain driving elements (not shown) for internal focusing
lens 4, for instance, two motors and the corresponding
linkage rods. Inasmuch as the construction of these driving
elements are well known, it does not seem necessary hera to
.
;- further aescribe them. It is only essential to point to the
fact that on the upper side of the housing 24, as Fig. 4
-:`
'
.
. :: - . .:
,
,, ` "' ~ ` .
shows, a display 25 is provided to indicate the position of
internal focusing lens 4. Display 25 encompasses, a scan
pattern plate 26 the position of int~rnal focusing lens 4 is
then indicated, by point 27.
The drive of the internal focusing lens 4 can take
place in the most varied ways. For instance, it would be
possible to move the internal focusing lens 4 by one or more
stepping motors. It is preferred however, if movement oE
:. .
lens 4 is possible in such a way that scanning along line 18
a
in Fig. ~4 is possible. This can be accomplished in a
simple manner by the use o two direct-current motors which
provide movement of internal focusîng lens ~ in X and Y
directions and permit the rotational speed of these motors
to be independently variable.
In Figs. 5 and 6, a device is shown, which can be used
for the controlling tWQ such direct-current motors. This
device comprises a joy-stick 30, moun-ted in circular aperture
28 of cover plate 29 and which can move on all sides. ~he
movements of joy-stick 30, are transmitted, corresponding to
its direction and amount of deflection, to two potentiometer
31, 32 or equivalent control-elements for varying the input
to a respective servo-motor. From Fig. 6 r it can be seen
that the joy-stick 30 is attached to carrier 33, which is
rotatably mounted in pivotabla carrier 34 axis of rotation
for carrier 33 is normal to axis 35 oi carrier 34. Base
plate 36 supports pivotable carrier 34,
Carrier 34 is connected to the tap of potentiometer 32
along axis 35 so that rotation of carrier 34 about axis 35
causes corresponding change in the output of potentiometer
32. In a similar manner, carrier 33, acts on the control
- i1 -
element of potentiometer 31, which is supportecl by carrier
.. . .
34.
In this way, a device provides change of the rotational
speed of either or both of rnotors controlled by potentiometers
31, 32 corresponding to movement of the joy-stick 30.
According to Fig. 5 joy-stick 30 is positioned i.n aperture
28, which is essentially spherical. ~he microscopist
therefore has the ability to vary the speed and direction
of movement of the internal focuslng lens ~, by the direction
and ext~nt of deflection of the joy-stick 30 while:observing
... .
. through the ocular and lens 4 on the display~
.
'
:- .
. 4 . . .
: ~ . ;
:. :
~; . . . :
- , .
;: .
, ~ .
,
.
." ,
- 12 -
.. ; .
.
:~ ' - - :
'' ' . .
