JPS61194418A - Inversion type microscope with manipulator - Google Patents

Abstract

PURPOSE:To improve operability and the operation efficiency of bricking by operating plural hydraulic operation parts associatively through a transmission tube and providing a lower-limit stopper for a needle holder to a hydraulic operation part other than a hydraulic operation part which is associated with a focusing mechanism. CONSTITUTION:A sample placed on a stage 12 is put in focus through a collimation handle 22. Then, a hydraulic operation part 23 is coupled with the fine adjusting handle 22a of the collimation handle 22. A glass needle 16 is set. The rough adjusting handle of a hydraulic drive part 17 and the fine adjusting knob of a hydraulic operation part 26 are used in combination to lower the needle holder 15 and the tip of the glass needle 16 is set slightly above the focus position. Then, the fine adjusting joy stick of the hydraulic operation part 26 is swung slowly to its lower-limit stopper position. Further, the fine adjusting knob is rotated in this state to lower the glass needle 16 until its tip reaches the focus position. Thus, the glass needle 16 is set at the lower-limit position of the tip part, and consequently the glass needle is not broken in bricking and the operation efficiency is improved.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an inverted microscope with a manipulator equipped with a microphone 17 and a manipulator rake for performing various detailed operations on objects to be observed with the microscope.

PRIOR ART A conventional inverted microscope with a manipulator of this kind, as shown in FIG. 13, for example, has a stage 1 and a culture vessel 2 placed on the stage I and containing cells 2a therein.
, a condenser lens 3 that is provided above the stage 1 and includes a ring slit 3a therein, and an objective lens 5 that is attached to a revolver 4 that is provided below the stage 1 and has a phase plate 5a therein. , the glass needle 7a is fixed to the lower surface of the stage 1 via the mounting bracket 6.
The cell 2a in the culture container 2 is equipped with a micromanipulator 7 extending near the focus position of the microscope, and the cell 2a in the culture container 2 is punctured with the glass needle 7a of the manipulator 1 and placed in the culture solution. The device was designed to perform a so-called pre-soking procedure that infiltrates enzymes, etc. However, in the case of this conventional example, since the plurality of operation parts 7b of the manibulator turf were mechanically interlocked,
There was a problem that the structure of the interlocking mechanism became complicated, and the reliability and durability of the device were low. Since the position of each operating section 7b is also restricted, there is also a problem that the operability is poor. In addition, since the bottom of the culture container 2 and the surface of the cultured cells are generally not uniform, when the tip of the glass needle 7a is lowered to the focus position, the tip of the glass needle 7a may accidentally hit the bottom of the container 2, causing the glass t.
There was a problem in that the t7a was often damaged, and as a result, the work efficiency of purinoking was reduced.

Purpose: In view of the above-mentioned problems, the present invention provides a device with high reliability and durability, excellent operability, and eliminates damage to the glass needle during pre-soaking, improving work efficiency of plucking. The present invention aims to provide an inverted microscope with a manipulator that is dramatically improved.

An inverted microscope with a manipulator according to the present invention includes a needle holder that holds a glass needle, a hydraulic drive unit that supports and drives the needle holder, and a hydraulic drive unit that is connected to the hydraulic drive unit via a hydraulic transmission tube. A micromanipulator is constructed from a plurality of hydraulic operation sections, one of the hydraulic operation sections is linked with a focusing mechanism, and the plurality of operation sections can be linked together simply by connecting them with a hydraulic transmission tube. The structure of the interlocking mechanism is simplified, and the reliability and durability of the fruiting device are increased. Furthermore, since the hydraulic operating section is connected to the hydraulic driving section only through the hydraulic pressure transmission tube, the hydraulic operating section can be placed at any position relative to the microscope body, resulting in excellent operability. . In addition, if a lower limit stopper of the needle holder is provided on at least one of the hydraulic operating parts other than the hydraulic operating part linked with the focusing mechanism, and the lower limit position of the tip of the glass hook is set to the focus position, the lower limit position of the needle holder can be set when the lowering operation is performed. The tip always stops at the focus position, and the glass needle also moves up and down in conjunction with focusing, so that the lower limit position of the tip is always set at the focus position, even if you refocus. The tip of the lens always remains in focus.

Hereinafter, the present invention will be described in detail based on an embodiment shown in FIGS. 1 to 9. FIGS. 1 and 2 are a side view and a front view, respectively, of an inverted microscope with a manipulator according to the present invention. In the figure, 10 is the microscope main body, 11 is the illumination system support, and 12 is the microscope main body 10.
As described in the specification of No. 55, a stage 13 is detachably fixed by a female dot/male dot structure, and 13 is a stage 12.
Above the lighting system support 11. 14 is a condenser lens supported by the condenser holder 13; 15 is disposed between the stage 12 and the condenser lens 14, and has a centering mechanism and holds the glass needle 16 in a hanging state. needle holder, 1
Reference numeral 7 denotes a hydraulic drive unit which is screwed and fixed to the upper surface of the stage 12, supports the needle holder 15 in a removable manner, and drives the needle holder 15 in the vertical direction, for example, as described in Japanese Patent Publication No. 57-28122; and is connected to the glass needle 16 via a hydraulic pressure transmission tube 19. An operation in which pressure is applied to the chemical liquid, etc. in the glass needle 16 by filling the hydraulic pressure transmission tube 19 with silicone oil, etc., and pushing out the medical liquid from the needle tip. This is an injector that does this.

Reference numeral 20 is mounted on the microscope body 10 below the stage 12 so as to be movable up and down. A revolver 121 is an objective lens fixed to the revolver 20. Reference numeral 22 is an objective lens fixed to the microscope body 10. A pair of focusing handles 23 are connected to the revolver 20 through a coarse and fine movement device as described in the above publication, and are detachably fixed to the microscope body 10 by means of mounting screws 24, and their operating portions are connected to the focusing handles 22. For example, it is connected to one fine movement handle 22a and is driven by the fine movement handle 22a.
A hydraulic operating section as described in Japanese Patent No. 7-28122, 25 is a hydraulic transmission tube connecting between the hydraulic operating section 23 and the hydraulic drive section 17, and 26 is connected to the hydraulic operating section 23 via a hydraulic transmission tube 27. This is a hydraulic operation part.

Furthermore, each part will be explained in detail.

3 and 4 are a partially cutaway plan view and a vertical sectional view of the needle holder 15, respectively, and 28 is a male dovetail 28 thereof.
a is an annular support frame that fits into a female dovetail of the hydraulic drive unit 17 and is fixed to the female dovetail by tightening a clamp screw 29; The centering knob 32.3 is pressed toward the center by the spring set 31, 3 screwed onto the support frame 28 and the centering knob 32.32.
An annular centering frame in which centering is performed by advancing and retracting 2, 33
is fitted into the centering frame 30, passes through the support frame 28, and is fixed to the centering frame 30 by tightening the fixing screw 34, which is screwed into the centering frame 30, and is fixed to the centering frame 30 by loosening it. An annular holder frame 35 that can be detached from the holder frame 35 has a base penetrating and held in the holder frame 33 in a horizontal state, a chiseled tip bent downward at the center of the holder frame 33, and a hydraulic transmission tube 19 attached to the base. It is an L-shaped holder to which a glass needle 16 is connected via a clamp screw 36 to the tip thereof.

5 and 6 are a partially cutaway plan view and a longitudinal sectional view, respectively, of the hydraulic drive unit 17, in which 37 is fixed to the upper surface of the stage 12 with screws and has a vertical female dovetail 37a. The mounting base 38 is pivotally connected to the mounting base 37 through the mounting base 37, and a head 38 for preventing removal having a groove is formed at once, and a spring 39 is fitted to the other end, and then a coarse adjustment handle 40 is fitted.
is the pinion shaft to which it is screwed. Note that when the pinion shaft 38 is fixed and the coarse movement handle 40 is rotated in the direction of screwing in, the rough spring 39 is compressed and a large repulsive force is generated, which is used to integrally move the pinion shaft 38 and the coarse movement handle 40. You can adjust the operating force when moving. Furthermore, the frictional force between the pinion shaft 38 and the coarse moving handle 40 is set to be sufficiently large due to the frictional force generated by the countersunk spring 39, so that only the coarse moving handle 40 rotates with respect to the pinion shaft 38 and loosens. Nothing happens. 41 is a female ant 37
A cylinder 44 is supported within the cylinder 41 so as to be movable downwardly on the mounting base 37 via a male dovetail 42 that fits with the a, and is moved up and down by a coarse movement handle 40 via a rack 43 that engages with the pinion shaft 38. 45 is a piston rod whose one end is fixed to the rolling diaphragm 44, and 46 is mounted on the cylinder 41 via a roller guide 47 so as to be movable in the vertical direction. The lower end is the piston rod 4
5, and has a female dovetail 46a formed on its back portion to fit with the male dovetail 28a of the support frame 28 of the needle holder 15.

FIG. 7 is a longitudinal cross-sectional view of the hydraulic operation unit 23, in which 48 is a cylindrical tube that is fixed to the microscope body 10 coaxially with the focusing handle 22 and is detachably fixed to a ring 49 with a mounting screw 24. A frame, 50 is a head fixed to the end of the frame 49, 51 is a rubber member etc. which is pivotally attached to the frame 48 coaxially with the focusing handle 22 and is press-fitted with the fine adjustment handle 22a of the focusing handle 22 at one end. A transmission shaft, to which a transmission drum 52 made of an elastic material is fixed, and a transmission drum 51b with a keyway 51a parallel to the rotating shaft is formed at the other end;
54 is a cylinder screwed onto the head 50, and 55 is a pin 5 screwed onto the inner end of the cylinder 54 so as to be coaxial with the transmission shaft 51 and movable in the axial direction.
Reference numeral 6 denotes a screw shaft that is slidably fitted with the key 'a51a, 57 is a rolling diaphragm that divides the inside of the cylinder 54 into an oil chamber 54a and a vacant space 54b, and 58 is fixed to the rolling diaphragm 57 at one end and screwed to the screw shaft 55 at the other end. It is a piston rod that is inserted into the center hole of the piston and is brought into contact with the inner bottom of the piston.

Here, with one rotation of the fine adjustment handle 22a, the objective lens 21
is moved by 200 μm, while the rotation of the fine movement handle 22a is transferred to the transmission drum 52. Transmission shaft 51. The rolling diaphragm 57 is moved by the axial movement of the piston rod 58 via the screw shaft 55, and as a result, an amount of oil corresponding to the amount of movement of the objective lens 21 is transferred to the hydraulic pressure via the hydraulic pressure transmission tube 25. It is assumed that the setting is such that the needle holder 17 is moved by the same distance and in the same direction as the objective lens 21 by being applied to the drive unit 17 .

If there is air between the objective lens 21 and the glass needle 16, "1"
As explained above, the cells to be manipulated are culture medium,
In many cases, it is in a liquid such as water, and the front optical path length that moves the focus position in the liquid changes, resulting in a change in the working distance of the objective lens. becomes out of focus. If the change in optical path length in the liquid is Δl, then Δ7! /n (n is the refractive index of the liquid) is the change in the working distance of the objective lens 21. In order to compensate for this error, the thread pitch of the threaded shaft 55 is appropriately selected,
The ratio of the amount of movement of the objective lens 21 by the fine movement handle 22a to the amount of movement of the needle holder 15 is expressed as the refractive index n of the liquid.
This problem can be solved by selecting a nearby location.

FIG. 8 is a longitudinal cross-sectional view of the hydraulic operation unit 26, in which reference numeral 59 is a base, 60 is a hand rest fixed on the base 59 via a column 61, and has a long groove 60a formed in the center;
63 is a cylinder fixed to the side wall of the frame 62; 64 is a rolling diaphragm that divides the inside of the cylinder 63 into an oil chamber 63a and a vacant chamber 63b; The piston rod 66 fixed to the rolling diaphragm 64 is fixed on the base 59 and is slidable in the horizontal direction (in the direction of arrow C in FIG. 8), and is connected to the other end of the piston rod 65. A slider having a lever 66a whose head is partially spherical;
67 is a support frame screwed to the top wall of the frame 62 so as to be movable in the vertical direction; 68 is a recess 68a rotatably supported by the support frame 67 and fitted with the head of the lever 66a at the bottom.
69 is a cylinder whose lower end is fixed to the upper end of the ball 68, and 70 is a fine movement joystick whose lower end is fixed to the upper end of the cylinder 69 and whose middle part is guided in the direction of arrow B by a long groove 60a. .

The vibration of the joystick 70 causes the ball 68 to rotate, causing the slider 66 to slide. 71 is an oil chamber 69a and an empty chamber 69b inside the cylinder 69.
Rolling diaphragm divided into 72 and oil chamber 63a
and 69a, 73 is a piston rod whose lower end is fixed to the rolling tire flam 71, and 74 is a piston rod connected to one end of the joystick 70 around the central axis of the joystick 70 (arrow C in FIG. 8). This coarse movement knob is mounted so as to be rotatable in the direction (direction) and is connected to the upper end of the piston rod 73 via a screw shaft (not shown), and moves the piston rod 73 in the axial direction by its rotation.

Note that both ends of the hand rest 6o with a length of 1s6oa play a role as stoppers that determine the swing range of the joystick 70.

FIG. 9 is a schematic diagram showing the overall configuration of the hydraulic drive system described in item 1, in which the hydraulic drive unit 17 that drives the needle holder 15
has a cylinder capacity of 12 strokes, so
A stopper is provided on each hydraulic operating section 23 and 26 so that the total stroke of each hydraulic operating section 23 and 26 does not exceed 12 m to prevent damage to each rolling diaphragm. In the case of this embodiment, each hydraulic operation section 23 and 26
are connected in series to the hydraulic drive unit 17, but they may be connected in parallel. Further, as long as the total stroke of each hydraulic operating section can be set within a range that does not exceed the stroke of the hydraulic drive section 17, any number of hydraulic operating sections may be set. In addition, in this embodiment, the hydraulic operation unit 23 that is linked to the alignment of the pin 1 is connected to the fine adjustment handle 22a of the focusing handle 22.
It may be directly connected to the objective lens 21 and the vertical drive mechanism of the revolver 20.

Since the inverted microscope with a manipulator according to the present invention is constructed as described above, first, focusing is performed on the specimen placed on the stage 12 using the focusing handle 22. Next, the hydraulic operating section 23 is connected to the fine adjustment handle 22a' of the focusing handle 22.

Next, the glass needle 16 will be set. At first, the condenser lens 14 will get in the way, so please push it away or remove it.
to raise the support frame 28 of the needle holder 15 sufficiently upward. After attaching the glass needle 16 of an appropriate length to the holder 35 of the holder frame 33, the holder frame 33 is fitted into the centering frame 30 and fixed with the fixing screw 34.

Next, the coarse movement handle 40 and the coarse movement knob 74 of the hydraulic operating section 26 are used together to lower the needle holder 15 and bring the glass needle 16 slightly to one side of the focus position. At this time, if a 4x to 10x objective lens 2 is used, the tip of the glass needle 16 will appear as a white dot in the field of view. Here, by moving the centering knobs 32 and 32 back and forth to adjust the position of the centering frame 30, the glass needle 16
Bring it to the center of your field of vision. It would be even better if you could use the cross scale on the eyepiece to center it accurately. Next, the follower joystick 70 of the hydraulic operating section 26 is slowly swung to the lower limit stop position.

Further, in this state, rotate the screw knob 74 to lower the glass needle 16 and align its tip with the focus position.
If so, the entire setting is complete.

Next, regarding blinking of cultured cells, by swinging the fine joystick 70, it is possible to move the glass needle 16 downward by about 100 to 500 μm with the focus position as the lower limit. Furthermore, when the fine adjustment handle 22a is rotated to correct the focus position, the movement is caused by the hydraulic operation section 23.
The lower limit position of the needle holder 15 is also interlocked with the lower limit position of the needle holder 15 . Therefore, when performing blinking, even if the bottom of the culture vessel or the surface of the cell culture cell is not a uniform plane, the tip of the glass needle 1G will always descend to the position where the cells are in focus. Therefore, it is automatically prevented that the glass needle 16 is damaged by accidentally hitting the bottom of the container with the tip of the needle, and as a result, blinking can be performed one after another. Further, in this case, since the glass needle 16 is held by the annular needle holder 15 via the annular holder 35, the phase contrast microscope is not impaired between the condenser lens 14 and the specimen.

Figures 10 and 11 are a partially cutaway plan view and a vertical sectional view of another example of the needle holder 15, respectively, in which a transparent disk 75 is used as a centering frame instead of the holder frame 33 of the above example. 3
0, and the base of the boulder 35 is held in the central part of this transparent disc 75, and the length from the held part (base) of the boulder 35 to the needle tip of the glass T116 is becomes shorter. Also in this case, since the holder 35 is held by the transparent disk 75, the phase contrast microscope is not damaged between the condenser lens 4 and the specimen.

FIGS. 12A and 12B are a sectional view and a side view of a fine movement handle of another example of the hydraulic operating section 26, respectively, and these are the fine movement joystick 70 and the coarse movement knob 74 of the example shown in FIG.
Instead, a fine adjustment handle 76° screw shaft 77 and a coarse adjustment handle 78. The screw shaft 79 is used to drive the cylinders 63 and 69, and the fine movement handle 76 is connected to the cylinder 63.
A long groove 76a into which a pin 80 fixed to the side slides is provided, and both ends of the groove 76a are configured as stoppers.

- 4 forces □Results As mentioned above, the inverted microscope with a manipulator according to the present invention can be linked by simply connecting the plurality of hydraulic operating units 23 and 26 with the hydraulic transmission tube 27, so the structure of the interlocking mechanism is It is extremely simplified, and the reliability and durability of the fruiting device are increased. Further, the hydraulic operation section 26 is connected to a hydraulic transmission tube 27.
．． Since it is connected to the hydraulic drive section 17 only through the hydraulic control section 25, the hydraulic operation section 26 can be set at any position with respect to the microscope main body 10, resulting in excellent operability.

Furthermore, even if the bottom of the culture container or the surface of the cultured cells is not a uniform plane, if the lower limit position of the tip of the glass needle 16 is set to the focus position, the tip will always remain in the focus position when the lowering operation is performed. At the same time, even if the focus is refocused, the tip always remains at the focus position. Therefore, the glass needle 16 is not damaged during blinking, and the blinking work efficiency is dramatically improved.

[Brief explanation of drawings]

1 and 2 are a side view and a front view, respectively, of an embodiment of an inverted microscope with a manipulator according to the present invention, and FIGS. 3 and 4 are respectively partially cutaway plan views of the needle holder 15 of the above embodiment. 5 and 6 are a partially cutaway plan view and a longitudinal sectional view, respectively, of the hydraulic drive unit 17 of the above embodiment, and FIG. 7 is a longitudinal sectional view of the hydraulic operating unit 23 of the above embodiment, FIG. 8 is a longitudinal sectional view of the hydraulic operating section 26 of the above embodiment, FIG. 9 is a schematic diagram showing the overall configuration of the hydraulic drive system of the above embodiment, and FIGS. FIGS. 12A and 12B are a sectional view and a side view of another example of the hydraulic operating section 26 and a side view of the fine adjustment handle 76, respectively, and FIG. 13 is a conventional example. FIG. 3 is a partially cutaway side view of the example. 10...Microscope main body, 11...Illumination system support, 1
2... Stage, 13... Capacitor holder,
14...Condenser lens, 15...Needle holder, 16...Glass needle, 17...Hydraulic drive unit, 18...Injector, 19,25.27.
...Hydraulic transmission tube, 20...Revolver-12
1...Objective lens, 22...Focusing handle, 2
3.26...Hydraulic operation unit, 24...Mounting screw. 2nd Figure 22a l'+ Figure 3 Section 4 Utsubo Seven Fukuro - 6 Figure 12 (A)

Claims (2)

[Claims] (1) An inverted microscope with a manipulator equipped with a micromanipulator for performing various fine operations on the object to be observed, which includes a needle holder that holds a glass needle and a hydraulic drive that supports and drives the needle holder. and a plurality of hydraulic operation sections connected to the hydraulic drive section via hydraulic transmission tubes, forming a micromanipulator, and one of the hydraulic operation sections is interlocked with a focusing mechanism. An inverted microscope with a manipulator. (2) An inverted microscope with a manipulator according to claim (1), characterized in that a lower limit stopper of the needle holder is provided on at least one of the hydraulic operating parts other than the hydraulic operating part that interlocks with the focusing mechanism. .