JPS58155078A - Device for transporting culture container - Google Patents

Abstract

PURPOSE:To transport a culture container to a given operating position without touching it with hands from the outside, by providing the culture chamber of an automatic culture device with a rotary member, a member for placing plural containers, and an elastic means. CONSTITUTION:The transporting device 6 consists of the rotary disc 35 set on a base plate fixed in a culture chamber, and rotated and driven by a motor. The shaking member 41 is set by the supporting shaft 39 in a shakable way in such a way that the tip part of the supporting member 38 is engaged with an inverted U-shaped dent part, and is exerted by force caused by the elastic force of the leaf spring member 43 in such a way that its tip part is rotated upwards. Consequently, the placing member 42 is made shakable, and a specimen in a laboratory dish can be prevented from being damaged or destroyed by the damping action of the leaf spring member 43 even if somewhat impact may be produced when the laboratory dish is placed on the placing member 42.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is used in a culture container, more specifically, in a culture silver type, which automatically performs subculturing of cells in a culture chamber maintained at a normal atmosphere. The present invention relates to a culture container transfer device.

As is well known, living tissue IIk and cell culture techniques are
In all fields such as medicine, biology, pharmacy, agriculture, etc.
This is an essential basic experimental technique for conducting research at the cellular level. However, obtaining stable cell lines by subculturing living tissues and microorganisms is technically difficult.

First, single, nine-layered cells are placed in a culture container such as a petri dish or an inner culture bottle, and diluted by injecting a culture solution into the cells. Then, the cells to be cultured will float in the culture medium and be in good condition, so this culture container! Culture cells are grown in a predetermined atmosphere at a temperature of 37° C., humidity of 100° C., and an acidic gas concentration of 611). After a predetermined period of time has elapsed, the culture container is removed from the culture chamber, and the state of multicellular culture is observed using an observation means such as a hazy microscope. Once it is confirmed that the required number of cells have proliferated, the next generation culture system can be created without using clean pliers or other tools.

In the work of creating this culture system, first, the culture solution in the culture container is aspirated with a pipette and discarded.

Next, the cells remaining in the culture vessel are washed by injecting a washing solution consisting of diluted phosphate, etc., and then the washing solution is discarded by suctioning with a pipette or the like.

This washing step is performed to flush out the old culture solution tube and enable the #cumulant to function effectively in the enzymatic treatment and salting step described below. Next, in order to release the 9 cells that have implanted and proliferated on the bottom surface of the culture container from the bottom surface, which is the growth surface, an enzyme overlaying a proteolytic enzyme such as trypsin is injected into the liquid tube culture container. Let the enzyme act on the cells for one hour.

Next, after aspirating and discarding the enzyme solution with a pipette, a new culture solution is injected into the culture container, and this is stirred by repeating suction and evacuation multiple times with a pipette, etc.
The released cells are singulated and resuspended in culture medium. Then, a predetermined amount of culture solution containing a large number of individualized and resuspended cells is divided and injected into multiple new culture containers, and the insufficient culture solution is added to create the next generation culture system. The meeting is complete.

Next, the above-mentioned culture vessel containing the cultured cells to be cultured for the next generation is transferred again to the culture chamber maintained in a predetermined atmosphere, and the culture is continued by multiplying the cells.

Conventionally, biological tissues and cells have been subcultured in this way, but the subculture method is manual and has various drawbacks. The shrimp are subjected to the sequential operations at -
They had to be taken out of the culture room, and the p% cells were affected by changes in the environmental conditions, resulting in changes in the proliferative layer, lifespan, etc. In addition, if the cells are contaminated by contamination with bacteria, they can become contaminated, die, or undergo metamorphosis. As the cell ages, the proliferative layer, lifespan, morphology, etc. of the cells change, and the disadvantage is that it is not possible to perform standardized culture under certain conditions. Furthermore, if the cultured cells are pathogenic bacteria, etc.
There is a risk of biocontamination (biological contamination).
Safety fiK also had its drawbacks.

Therefore, in order to eliminate these various drawbacks of the conventional methods, it is necessary to automatically create a next-generation culture system in a 9-culture chamber maintained in a subliminal atmosphere, and to carry out subculture of cells [ It is desirable to use an automatic cell culture device that can perform cell culture continuously under certain conditions. However, such an automatic culture device has not been provided so far, and in order to manufacture it In order to create a next-generation culture system indoors, it is essential to have a transfer device that can move a culture container stored in a culture chamber to a predetermined operating position without touching it from the outside.

The object of the present invention is to solve the above-mentioned problems by including a rotating member driven by a motor in the culture chamber of an automatic culture device, and a plurality of rotating members attached to the outer circumference of the rotating member so as to be swingable. A container mounting member and an elastic means for biasing the container STM material to assume a horizontal position are provided, respectively, so that the culture container 11i can be transferred to the operation position without touching it from the outside. to provide transfer equipment for the device.

Hereinafter, the present invention will be explained based on an illustrated embodiment.

FIG. 1 shows an automatic cell culture device equipped with a culture container transfer device according to an embodiment of the present invention. This automatic culture device 1 has a constant atmosphere (for example, a temperature of 37° C.
In order to perform various operations on the petri dish 4 stored in this culture chamber 3 and kept at a humidity of Zoo * and carbon dioxide concentration of 5-), the culture chamber 3 is kept at a temperature of 5. It is configured with various 1 & positions attached. That is, the automatic culture device 1 includes the culture chamber 3, a loading/unloading device 5 for automatically loading and unloading the petri dishes 4 into and out of the cultivation chamber 3, and a loading/unloading device 5 for automatically loading and unloading the petri dishes 4 into and out of the cultivation chamber 3, and a device 5 for automatically loading and unloading the petri dishes 4 into and out of the cultivation chamber 3; A transfer device 6 for transferring the books to MlliK, an Is liquid device 7 for suctioning and removing the liquid that has become annual bonds from the Petri dish 4 yen, and a liquid supply device 8 for supplying the liquid necessary for culture into the Kuyare 4. , a peeling step 11jt9 in which the cultured cells are peeled off from the growth surface by applying vibration to the Petri dish 4, and a dispensing step for stirring the liquid in the Petri dish 4 or dispensing it into another new Petri dish 4. A device 11, a Petri dish supply device 12 for supplying a new Petri dish 4, a control needle aill that automatically controls the operations of the various devices mentioned above, and a rounding device for observing the culture *am' in the Petri dish 4 from the outside. observation device 1
4 constitutes the main expense department.

As shown in FIGS. 1 and 2, the loading/unloading device 5 is located near the center of the right 1111Il of the housing 3.
As shown in FIG. 3, the housing 21 of the device 5 is provided so as to communicate with the culture chamber 3 and the outside.
3, an inner chamber, an intermediate chamber, and an outer chamber are separated by an intermediate wall 22 formed in the right jar of the casing 2 and the housing 21.
It is divided into 1W7L. A belt conveyor 2 rotatably supported in the center by a support shaft 23m is installed in the above-mentioned inner chamber.
3, there is also a shutter 24 and a groove in the middle room.
Il @ closing solenoid 25 and belt;
Further, a shutter 27, a solenoid 28 for opening and closing the shutter 27, and a belt conveyor 29 are disposed in the outer room.

The upper ml belt conveyor 23 has an inner end extending through an opening 21a formed in the outer wall of the inner chamber of the housing 21 to reach the culture chamber 3, and connects the inner end of the housing 21 with the housing. 21 and the hook are rotated around the support shaft gas in the anti-saki direction by the tension coil spring 31 that is passed! 1 is given. The rotation of the conveyor 2B due to this behavior is regulated by the shutter 24 connected to the outside 111IlB of the conveyor 2s, which is bored on the right side of the housing 3, closes the nine openings 21, and stops at the nine position. has been done.

In this restricted position, the inner end of the conopia 230 corresponds to the lower part of the mounting member 42 of the transfer device 6 (see FIG. 4) of the present invention, which will be described in detail later. Below, conveyor 23
The position of the mounting member 42 that corresponds to the inner end of will be referred to as the 1 loading/unloading position. When the shutter 24 is turned on, the conveyor 23 is rotated in the direction of the hour hand against the elasticity of the spring 31. 9 was transferred and placed on the transfer device, with the inner end sK provided with nine matching claws! mb K hooks Petri dish 4 onto conveyor 23
Place it on top and carry it out towards the belt conveyor 26 in the intermediate room. It's on. The shutter in the intermediate chamber is slidably disposed so as to open and close the opening in an airtight manner, and the solenoid 3! It is opened and closed by +. Then, the opening 2a
The belt conveyor 2 is disposed at a height corresponding to the height position. In front, the shutter 27 inside the outer room is
Nine holes are drilled in the middle 922 and are slidably arranged to open and close in an airtight manner, and the solenoid 28
It is designed to be opened and closed by.

In this way, the intermediate chamber has both shutters 24. ! 7K, it can be opened and closed in an airtight manner, and serves as a ninth buffer chamber that does not allow direct communication between the culture 11s and the outside. By providing the intermediate layer, it is possible to prevent sudden changes in the environmental conditions within the culture chamber 3, and it is also possible to prevent bacteria and the like from entering the culture chamber 3 from outside.

The above-mentioned peltco/pair 29 is disposed at a height position corresponding to the above-mentioned opening 228K, and the same conveyor 2
The outer end of the housing 81 corresponds to the tray 32 that is exposed to the outside through a hole 21b formed in the outer wall of the outer chamber of the housing 81.

According to the loading/unloading device 2 configured in this manner, each belt pair 23, 26 is
, the 290-drive low two was assisted in the direction of the anti-saki meter, and the solenoids gs and ms were energized at the appropriate time to trigger the shutter! 7. By moving 24t$1w11, the petri dish 4 fed from above the tray 32 is transferred to the conveyor 29,
2@ and 2B, and the petri dish 4 is automatically transferred onto the transfer device 6 by the conveyance force. Also, when transporting petri dish 4, each Peltoco/pair 2B
, 214 , 29 6-drive field-2 is rotated clockwise, and the solenoid 912! s, timely energize 2m and shutter ha, 27tllllf! By doing so, the Petri dish 4 on the transfer device 6 is hooked by the engaging claw 23b and placed on the conveyor 8s, and then the conveyor! B
, 2@, and 29, and the tray 3
Petri dish 4 is taken out on top of 2.

Note that, as will be described in detail later, the mounting member 42 of the transfer device 6
A cutout of 4 mb is provided at the inner end St* of the belt conveyor 23 so that rotation of the belt conveyor 23 is not inhibited by the mounting member 42 (see II in FIG. 4).

As shown in FIG. 1, the transfer device 6 representing one embodiment of the present invention is composed of a rotary table placed on a base @SS provided in the third circle of the culture chamber, and is driven by a motor 34. 〉It is designed to be rotationally driven.

That is, the transfer ate transfers the substrate 3 as shown in FIG.
3, an output gear 36 is meshed with a nine-rotating disc 3s which is planted and rotatably supported by a nine-support shaft aSS, and a nine-toothed gear ssb carved on the outer periphery of this rotary disc 3s. ,
The motor 34 that rotates the circular plate 31st, the 9 plurality of petri dish placement parts 1i537 which are attached to the peripheral edge of the rotating circle @3S and arranged at equal intervals, and the movement of the petri dish placement part 37. Round optical sensor f-44 for position detection, rotational position and initial position.
1,44b, the main lIs is constituted by the petri dish mounting section 37k! , as shown in Figure S.
t has an arm-shaped 5 fixed to the rotating disk 111 s#3
8, a swinging member 41 having an inverted U-shape in plan view, which is swingably attached to the tip of this support *s*tis by a horizontal support shaft 39; A loading member 42 is fixed to the tip end step and has a nine-ring shape with a part cut out, m- is fixed to the lower surface of the support member 38, and another one is fixed to the lower surface of the swinging member 41. A double leaf spring member 48 and a light reflecting member a for detecting the rotational position and initial position attached to the outer peripheral surface of the tip ■O of the primary mounting member 42
ha, 45b. Note that the light reflecting member 48b for initial position detection is
It is provided only in one petri dish mounting section 31 out of 7. The first half vibrating member 41 is pivotally connected to the oscillating member by a support shaft 39 with the tip S of the support member 38 fitted into an inverted U-shaped recess. St- Receives an urging force that causes it to rotate upward. The rotation of the swinging member 41 due to this biasing force is normally kept at a level tec*m by the standard hand restraining hand RK to keep the mounting member 42 in a horizontal state. The reason why the plate member 43 has a double structure is to maintain durability and flexibility so that the elasticity does not deteriorate even after long-term use. The mounting member 48 is provided with a protrusion 42a f having an inner diameter slightly larger than the outer diameter of the bottom surface of the petri dish 4 on the outer part of the upper surface so that the petri dish 4 placed thereon does not fall off during transfer. 9. The partial notch tiII is formed so that the circle gaII of the belt conveyor 23 of the carrying-in/unloading device 5 can fit into the rotating part, and the petri dish 4 can be placed or removed. This is formed with nine notches 42b inclined at a constant angle with respect to the longitudinal direction of the mounting portion 37.

As mentioned above, the placement member 42 is arranged so as to be able to swing freely while being biased by the leaf spring member 4s. This is to prevent the sample in the petri dish from being damaged or broken due to the f-pressure action of the leaf spring member 43 even if the sample is broken. 9. When suctioning the chamber inside the petri dish 4 using the waste liquid device 7 (see FIG. 6) or the dispensing device 11 (see FIG. 9), which will be described in detail later, the tip s8 or the pipette 97
This is to prevent damage to the petri dish 4 by absorbing the impact when the suction end of the plate collides with the bottom surface of the petri dish 4.

Furthermore, in this case, the tip 58 or the suction end of the pipette 97 presses the bottom surface of the Petri dish 4 to perform suction in a good condition, and the bottom surface of the Petri dish 4 is slightly tilted so that the suction end and the bottom surface of the Petri dish 4 are It is rounded to form an opening in between and to bring the liquid gold to the side so that the suction of the liquid is complete and effective.

The optical sensors 44a and 44b are arranged at a height that can face the light reflecting members 45a and 45b, and receive the light emitted by themselves after being reflected by the light reflecting members 45 and 4Sb. The position of the Petri dish W637 is detected. This optical sensor 44a
, 44b are both connected to the control device 13, and the control device 13 connects both optical sensors 44a, 44b.
The rotation of the motor 34 is controlled based on the output of the petri dish mounting section 31 to move it to an appropriate position.

In addition, in FIG. 1 and the fourth button, the reference numeral 46 indicates the rotating disk 3.
The same disk 3 is pressed against each of the three equal parts of the outer peripheral surface of
It shows a 3fi ga 4 troller that smoothly regulates 50 rotations.

As shown in FIG. 1, the waste liquid device 7 has a main body part housed in a machine room 41 provided above the culture chamber 3 in the housing 2, and the main body part is housed in a machine room 41 provided above the culture chamber 3. Through 48,
The bottom part extends 3 circles I/c of the culture medium.

As shown in FIG. 6, this liquid waste device 7 includes a liquid drain pipe 51 whose upper base end is fitted into one end of a liquid drain pipe 49 connected to a suction pump (not shown), and a liquid drain pipe 51 connected to a suction pump (not shown). A support member 52 that holds the intermediate tube 51 by fixing it near the upper base end of the liquid tube 51, a drive belt 53 that fixes the support member s2 to a part of the circular surface, and this drive bell)! The Jt hook is inserted into the pair of upper and lower pulleys 548 and 54b, the motor 55 attached to the output shaft of the lower pulley 54b, and the guide hole drilled in the support member 52 to move the member 12. It is formed of a guide bar S that regulates the water in the vertical direction, a support 41I device 7 that supports the pulleys 6411, i4b, the guide bar 6@, etc., and a short cylindrical tube that serves as a suction nozzle at the tip of the drain pipe s. chip ist
@The chip storage container i9 to supply, this chip storage container S, the storage device support plate 1 to hold the meeting, and the chips stored in the chip storage container 11! a chip supply control device 63 for controlling the supply of chips 68; a chip supply control device 3 connected to the chip storage device s9; a solenoid 64 for roughly driving the chip supply control device/st;
Penetrating the ceiling 114- of the above culture 1 ms, the same 1148
The main lIs is composed of a round guide tube 2 whose upper end has an outwardly projecting edge, and a tip insertion/removal tube 6 attached to the lower end of the guide tube l.

In this m-liquid device I, the chip station delivery device Se, chip supply control device-3, chip insertion/removal tube, etc. are installed so that the lower end S of the drain pipe ji1 can be directly inserted into the petri dish 4. This tip S will enter the KI[II in different dishes 4, which will cause cross contamination between the Petri dishes 4, so a tip is placed at the tip of the drain pipe s1. I installed the 58t and replaced it every time I drained the l-.

The chip accommodating container 59 is, for example, a short cylindrical chip having a diameter of IIII and a length of 1 to 1 lll.
The lower part is formed of a conical cylindrical body, and the upper end opening is provided with an i [III that can be opened and closed freely, and the lower end opening is Chip Gaitochi - Connected to Pro 3. and,
This chip storage s9 is identified and supported at the upper end of the storage support I9 @ 61, and is vibrated by a lifting device (not shown), vibrates together with the thin support plate 61, and the chips 5IItlI11 Chip Gaitochi, -
163, it is sent to the chip supply control device 62.

The chip supply control device 68 is mainly composed of a cylinder member 67 and a piston member 68, and the piston member 6 is designed to prevent it from colliding with the drain pipe 51 when the member @8 moves. Ninth play hole, 8 holes, and tip 511
IH chip storage hole for storing and sending out one chip at a time
b are provided so as to penetrate in the vertical direction, respectively. The piston member 68 is given a rightward sliding habit by an extensible coil spring 69.
At the position moved toward the right, the cylinder member 67
The tip guide attached to - the other end of Pro 3;
One chip 58 is accommodated in the inner hole ssb circle in correspondence with the chip accommodation hole 68b. Further, when the solenoid 64 is energized, the piston member 68 connected to the plunger 6414C of the solenoid 64 is moved toward the left against the elasticity of the spring 6g, and the chip storage hole 68b is closed to the guide tube 5. A PC is provided so as to accommodate the nine tips 58 in correspondence with the nine tip guide holes 65b drilled in the tip and supply the nine tips 58 to the tip insertion/removal tube 68K.

The guide tube 6s is formed of a cylindrical body having an outwardly projecting edge at its upper rim, and has a central hole serving as a drain guide hole SSa through which the drain tube 51 is inserted.

Further, the tip guide hole 6sb is bored parallel to the drain pipe guide hole 651. The tip insertion/removal tube 66 is made of an elastically deformable plastic material. It has a changing structure. Then, from the small diameter part to the middle of the middle diameter part, a cross-shaped sliding part 661 is inserted when viewed from the horizontal side, and furthermore, on the outer periphery of the upper part of the outwardly protruding edge of the f tip 1llllllll. is a contractible ring-shaped coil spring 7
1 is installed. At this ninth point, the sliding part 66m is closed by the elasticity of the spring 71, and the ninth tip 58, which is supplied through the guide hole ssb, is guided by the conical part until it reaches the middle oneills. The position is regulated by the small 1111 in the middle part, so that a waiting state is taken.

Note that the arrangement position of the mil [device 7 is as shown in 4th aI, the lower tip of the drain pipe 51 is one of the mounting members 4 in the transfer device 6! The moth is also spaced apart from the support shaft 39 so as to correspond to the outer peripheral edge of the support shaft. Hereinafter, the position of the corresponding mounting member 48 of the drain pipe s1 will be referred to as the 1 waste liquid position.

The distal end portion is formed so that it is easy to fit the tip s8 in the circumferential direction, and also prevents the tip s8 from fitting into the drain pipe 51 more than necessary. Further, the outer diameter of the drain pipe 51 is selected so that the outer diameter of the tip 58 is also slightly smaller.
When the tip 58 moves back upward, the upper end surface of the tip 58 abuts the lower end surface of the tip insertion/removal tube 66, and the tip 58 also automatically falls off from the drain tube s1.

The thus constructed waste t4i1t7 is placed on the transfer device 6 and moved to the waste liquid position into the nine petri dishes 4.
To remove unnecessary liquid from inside, first remove solenoid 6.
The piston member 68 is slid toward the left by driving the chip storage hole 61b and the tip guide hole 65b.
In correspondence with K, the chips 1s8t- and chip guide hole 6, which were previously stored in the chip storage hole 68b,
When the tip s8 is supplied to the tip insertion/removal tube 66 by its own weight through the 5"bt, the tip s8 is in a state where the slot 416a is closed in normal life, so the inclined step surface between the medium diameter part and the small diameter part. The motor 5S (-) is driven to move the belt 53t, and the belt 53t is moved to -2.
* Lower the member 52 downward. Then, the drain pipe 51 fixed to the member 52 descends together, and the g
ss enters the tip insertion/removal tube 66FF3 from the drain pipe guide hole aSa circle of the guide tube 65. Then, it comes into contact with the tip s8 which is set in the timing state at the middle diameter position, and the small diameter part of the tip of the drain pipe 51 fits into the center hole of the tip 58, so that the drain pipe 51
1, the chip 58 is tightly sealed. When the drain pipe s1 further descends, the tip 58 is forcibly entered into the small diameter portion, and the sliding portion 66a is pushed open against the elasticity of the spring 71, increasing the inner diameter of the small @ portion. Therefore, the drain pipe Jl forcibly enters the small diameter portion, protrudes from the distal end of the tip insertion/removal tube 66 with the tip S8 attached thereto, and continues to descend further. Then, the tip aSt of the chip b8 is brought into contact with the outer peripheral part IFJ) of the bottom wall of the petri dish 4 which is in the state of being released at 4at-111 by the container closing device 78 (see 7th @) to be described later, and the tip aSt of the spring 43 is The swing s@41 and the mounting member 421 are rotated against the elastic force, and the petri dish 4t is slightly tilted, and the motor 55 is stopped to stop the drain pipe 51 from descending.

Next, a suction pump (not shown) is driven, and the liquid in the Petri dish 4 is transferred to the chips 58. Drain pipe 51. Drainage tube 49
t-through to a waste tank (not shown) in the W7T leg. At this time, since the Petri dish 4 is pushed by the chip 58 and is tilted, the liquid in the Petri dish 4 gathers near the opening at the tip of the chip s89, and the liquid flows into the Petri dish. 4- It is all suctioned and discharged without leaving any residue inside.

When the suction pump is operated for a certain period of time and the liquid in the Petri dish 4 is completely discharged, the motor 55 is then rotated in the opposite direction to the previous direction, and the liquid drain pipe 61 starts to move upward. Then, first, the petri dish 4. which was being pushed through the chip 5B. Placement member 42 + **WA material 4
1, N' positive pressure is released 1t, and the spring 43 returns to the horizontal position. Subsequently, until the lower end surface of the chip 58 abuts the lower end surface of the chip insertion/removal tube 66, the drain pipe s1
When the tip 58 rises, the tip 58 cannot rise any further due to the collision, and enters only the drain pipe 51 or the tip insertion/removal tube 66, and the tip 58 and the drain pipe 51 are separated. When the fitting is removed, the chip 58 falls off due to its own weight and is automatically stored in a chip storage tank (not shown) outside the culture chamber 3. Needless to say, the used chips 58 that fall off during plating are prevented from falling into the middle of the petri dish 4 by means of suitable guide means.

The liquid supply device 8 supplies cleaning liquid into the petri dish 4.

As shown in FIGS. 1 and 2, there are storage containers 72a and 72b for storing the cleaning solution, enzyme solution, and culture solution, respectively, as shown in FIGS. 1 and 2. , ? +ac, and a cooling storage tank 7 in which these storage containers 72m, 72b, 72Ct- are stored and stored at a low temperature of, for example, 1°C to 4°C.
3, and a ninth roller pump 74m that supplies each liquid in the storage containers 72a, 72b, and 72c to the petri dish 4.
, 74b.

74C and this roller pump 74a? 4b,
The liquid sent out by 74c is heated to the same temperature as the atmospheric temperature of culture chamber 3 (for example, 37°C) by heating a 7J+1, 7Sb, '76c, and the liquid position of each of the above liquids is guided. Liquid supply chip 741m, 7
6b and 76c.

Like a normal refrigerator, the cooling storage tank 7s cools the ellipse by utilizing the heat absorption/heat generation phenomenon during expansion and compression of the refrigerant, and the temperature inside the tank is detected by a temperature sensor (not shown). Based on this output, the compressor (@l
(not shown) is operated to maintain a constant temperature. This cooling storage tank 7s is used to store the culture solution in enzyme solution for a long period of time under high temperature conditions.
Enzymes in the solution (e.g. trypsin) are inactivated99, and the culture
Since the vitamins and amino acids in the liquid may become deactivated, in order to prevent this, each of the above-mentioned tubes are provided to be cooled and stored.

The above roller pump '14m, 74b, ? 4C
A rotating roller (all not shown) and a liquid conveying device.

The main body is composed of an arm (not shown) that presses against the roller.
The liquid in the liquid feeding tube is fed by 1 g of 1 transfer. This roller pump 74a.

? 4b, 74C depends on the rotation speed of the above roller,
This has the advantage that the amount of liquid supplied can be accurately controlled. Mayu, the above warmer 10.75b? The IC includes a heater (not shown) and a temperature sensor (not shown).
It has a built-in temperature sensor and controls the power supply to the heater based on the output of the temperature sensor. y
sa, yib, 7Sc function to raise the temperature of the t411 body to a temperature similar to that of the culture chamber 8. This warmer 7! I11, 71b.

If 75C is cooled and stored in the cooling storage tank 7s and supplied to the petri dish 4 in the direct culture chamber 3, the cultured cells in the petri dish 4 will die or metamorphose due to a drastic change in temperature conditions. , rounding is provided to prevent this.

In addition, as shown in Figure 1161 and Figure 114, the supply tm of the above liquid supply tubes tea, t6b, and ysc.
is inserted into the culture 113 through the ceiling 1i48Q, and corresponds to the center of one mounting member 4 in the transfer device 6. Below, this chip-7
Placement member 4 directly below 41a, 76b, 741c
Position 2 will be referred to as the liquid supply position. In addition, the above storage containers 72g, 7Sb, ? 2c includes the 21st C
As shown in I, a ventilation filter 77 of 02μli degree
a, 7fu b * 7? C is attached to the container 9, 72m and 72b respectively.

The storage and feeding of the culture solution etc. is carried out in a safe manner by preventing the contamination of various germs etc. when air flows into the 72C.

In addition, as shown in FIG. Liquid silver amount, liquid supply and waste 111 IK petri dish 40 @ 4 m
Round-shaped lid opening WAil positions 7I for opening and closing are respectively provided. Jin's lid-1! As shown in No. 711, the fJlj position 78 includes a pair of rounded left and right arm members 79 @1 that hold the Petri dish 401141 from both sides, and a right arm member 850 St that are integrally fixed and driven by 9. m wheel 82 and an output wheel 5at meshed with this drive gear 82
- One end S is fixed to the upper surface of the motor 84 attached to the output shaft and the base of the right arm member 81, and the other end S is fixed to the left side perpendicular to the right arm member 81.
is implanted in the extended connecting member 85 and the bulge end of this connecting member 8s, and inside the left arm member 7.
and the arm member 7G.
, 'The hook is passed near the tip of @1, and the left arm member 79 is pivoted so that the tip of the left arm member 79 returns to the tip of the right arm member 81. a tightening coil spring 87 that biases the coil spring 86 to rotate in the circumferential direction of the arm member 86, and an adsorption piece 88 made of a strong material fixed to the right side surface of the rear end of the left arm member 790. , the electromagnet 8 disposed so as to face the adsorption piece 88 in a state in which it has been rotated to a position (the position shown in FIG. 7) where it can clamp both the earth alloy materials 711 and 81'll5114m.
It is composed of 9. Note that both arm members 79
, 111 have partially arcuate notched surfaces 791, 81 on the inner side of the tip of the lid 41t'' to make it easier to hold the lid 41t''.
m are formed respectively.

Further, the rotation of the arm member 79 due to the elasticity of the spring 87 is
member 79, 81F) l14at- at the tip l1lIiS
It is regulated at a position that is sufficient for clamping and where the attracting magnetic force by the electromagnet 119 can effectively act on the attracting piece 88.

When opening the petri dish 4 with the lid opening/closing device 1s configured as described above, first, the motor 84 is energized and the motor 84 is rotated counterclockwise.
The drive gear 82t and the support shaft 82M are rotated in the hour hand direction through the output gear 83. Then, the arm portion $ill, which is normally placed in the upright position, is rotated clockwise together with the drive gear 82. Finally, the arm member 79 is also moved from the upright position to the horizontal position together with the arm member 81 via the connecting member 86. And both arm members 7
At the stage where 9 and 81 are in the horizontal position, electricity is started to be applied to the electromagnet 89. Then, the adsorption piece 88, which had moved to a position facing the electromagnet 89,
The arm member 79 is pulled by 9 Klk, and the arm member 79 is rotated around the support shaft 86 in the seven-turn direction against the elasticity of the spring 87, and the tip *S of the arm member 79 is at the position where it collides with the upward direction of i4a. Then, as the motor 84 continues to rotate, the shear members 79 and 81 move to the horizontal position, the power to the motor 84 is cut off, and the arm member 71
1*81 is stopped in a horizontal state, and the electromagnet 8
Power to 9 is also cut off. Then, the restraint due to the attractive magnetic force of the electromagnet 89 is released, and the nine arm member 79 is moved by the spring 87.
0 elasticity rotates clockwise around the support shaft 86, and the arm members 79, 81 are rotated clockwise on both sides 79a, 811.
Attach a 4m long ryojo mask and hold the same amount of 4a and f.

At this time, since there is a slight barrier between the ridge 4 of the mounting member 42 of the transfer device 6 and the petri dish 4, the plate 44m moves slightly together with the petri dish 4, and both arm members 79,
It is firmly held by at. After the lid 4m is held between the arms S members 79 and 81, the motor 84 is energized again, and this time it is rotated clockwise in the opposite direction.

Then, the drive gear 82 is rotated counterclockwise via the output gear 8st, and both the arm members 79 and 81 are moved to l1Aa.
Rotate counterclockwise while holding it in place.

When both arm members 79 and 81 are moved back to the upright position,
% -1-84 is de-energized, the motor 84 stops, and the opening operation of the bag 4a is completed.

Next, when opening 1 and closing the 9 f14af by placing it over the Petri dish 4 again, first, in the same way as in the opening operation, energize the motor 84 to rotate the motor 4 counterclockwise once, 1 in the clockwise direction of the drive gear 82 via the output gear Is.
Turn it around. Then, both the arm members 79 and 81 move toward the horizontal position, so that they reach the horizontal position and at the 9th point, the basket is turned off.
motor 84 [- When stopped, both arm members 79, a
ll is covered with the lid 4at to be clamped on the petri dish 4, and the movement is stopped for 1Ibt in good condition.0Next, when the electromagnet 89 is excited, the adsorption piece 88 located at the position opposite to the electromagnet 1i9 is attracted, and the arm member 79 is Against the elasticity of spring 87,
The circumference 9 of the support shaft $6 is rotated counterclockwise, and the tip of the arm member 79 moves away from the side Kli from the position where it contacts the gA direction of the lid 4a. Next, rotate the motor a4f in the clockwise direction opposite to the front fence, both arms $179, @
1#ii4 Begin moving towards the standing position without holding for 1m.

At 70, when both arm members 7G and ill move to a position where they cannot be held again, the electromagnet @e
When demagnetizing, the arm member 79 is rotated in the hourly direction around the support shaft 86 by the elastic force of the spring s7, and is stopped at a predetermined position by a regulating means (not shown).

Then, when both arm members 79 and 81 return to the upright position, the rotation of the motor 84 is stopped and the closing operation of the chisel lid 4m is completed.

The peeling device 9 is disposed on one side of the transfer device 6 in the culture medium 11m3, as shown in the 111th & s4gec, and the first one is intermittently connected to the power supply device (not shown) as shown. A solenoid 91 that generates m- by being energized with current, and a support member 9 that supports this solenoid 91 [-
8, a nine-stroke member 9s attached to the tip of the plunger 91a of the solenoid 91, and a petri dish holding member fixed to the support member 92 and located directly above the petri dish 4 placed on the transfer device 6. It consists of 94. This peeling device 9 cannot sufficiently peel the enzyme II and cultured cells after passage from the growth surface of the petri dish 4 only by the stirring operation using the dispensing device 11 (see Figure 1) detailed in *. , is provided in order to apply lateral vibration to the petri dish 4 and ensure that it is detached from the cell culture growth surface.

The support member 92 is bent into a crank shape when viewed from the side, and its base end is fixed to the substrate 11. The solenoid 91 is attached to the intermediate portion of the support member 92 in the -vertical direction.

This solenoid 91 is attached to the supporting member so that the plunger 9111 is passed through the opening 92a formed in the support S member 92 and in good condition, the tip of the plunger 91M faces the -II1 surface of the petri dish 4 on the transfer device 6. At the tip of the plunger 911 fixed to the plate 92, the above-mentioned hitting member 9s made of a material such as plastic or rubber that will not damage the Petri dish 4 even if it hits the Petri dish 4 is attached. Further, the distal end of the support portion #92 extends horizontally to the upper side of the petri dish 4, and the petri dish holding member 94 made of a material such as plastic or rubber is fixed to the lower surface thereof. The solenoid 91 is provided with an adjustment screw 91b, and by rotating and adjusting this screw 91b, the vibration force tl applied to the petri dish 4 can be adjusted.
It is now possible to ill.

With this structure, the detachment device 9 applies vibration to the petri dish 4, and first, an electric current is intermittently applied to the solenoid 91 during the rounding process to detach the cells.

Then, the plunger 918 reciprocates in the left-right direction with the current energization cycle, moves leftward and hits the side of the petri dish 4 or the side of hs@4m at the 9th position with the tip of the hitting member 93. The Petri dish 4 is moved rapidly within the range allowed by the protruding edge 411 of the mounting member 411, and
vibrates periodically. Due to this vibration, the cultured cells in a free state are gradually peeled off from the bottom surface which is the growth surface of the petri dish, and if the vibration is continued for about 1 minute, the cultured cells are completely peeled off from the bottom surface. During this peeling operation, 1141m may receive a force in the direction of coming off from Kuyare 4 due to vibration, but since there is a holding member 94 directly above %114a, there is no risk of the lid coming off.Therefore, the peeled cultured cells , there is no risk of it flying out of the petri dish 4.

As shown in No. tg, the dispensing device 11 has a main body disposed inside the machine 1141, and the culture chamber 30 ceiling 4.
48 (-The dispensing operation part is drilled out into the culture chamber 3 by penetrating it. This dispensing device 11
A pipette 97 (see FIG. 9) was used to aspirate and eject a plurality of round bamboo-shaped petri dishes 4 supplied from a petri dish supply device ls (see FIG. 13), which will be described in detail later. The pipette supply device 195 (10th! Ii1 reference bank) and the pipette removal device 196 (1st
(See Figure 1) is attached. In this way, the reason why the pipette supply device 95 and the pipette removal device 96 are attached is that when one pipette 97 is used to stir and dispense the culture solution in a plurality of petri dishes 4, it is possible to Because there is a risk of cross-contamination, pipette 9 should be used for stirring and dispensing the culture solution in one Petri dish 4.
It is rounded so that 7ft is exchanged.

The above dispensing [1111 is as shown in the ninth fat, suction 49
Pipette on 8a) 97ti 1st Perose Bomp 98
and a rotating arm 99 with a bellows pump 98 mounted on the free end i1.
A rotary and sliding shaft 101 fixed in the direction of the base tm, a bearing member 102 that rotatably and slidably supports the rotary and sliding shaft 101, and a bearing member 102 that is attached near the upper end of the rotary and sliding shaft 101. A driving gear 101, an output gear 104 meshed with the driving gear 103, a rotary driving motor 105 fixed to the output shaft of the output gear 104, and a plunger TOSS 1 connected to the upper end of the rotating sliding shaft 101. The nine sliding solenoid 1018 and the plunger 10
611, and the above-mentioned (b) rolling and sliding shaft 101
Its main portion is composed of an extensible coil spring 107 that biases downward.

The bellows pump 98 is already well known, and includes
It has a short cylindrical outer shape, and a tubular suction 4981 projects downward from the center of its lower end surface. A collar @98C is provided in the middle of the suction end g81, and an extensible coil spring 108 is wound between the collar 98C and the pump 98. As shown in FIG. 11, this coil spring 10B is configured such that a bellows pump 98 is provided at the tip gIAs of the rotary arm 99 and is detachably attached to the rotary arm 99 by fitting the suction end into the notch 991. Therefore, the proximal end s' of the suction end 91151 is fitted to the axis of the cut 9, and the bellows pump 9- is attached to the tip of the rotary arm 99. It serves to firmly identify the bellows pump 9811ii1 rotation arm 99 by biasing it in the t-phase direction away from the trout@esc. Also, when the pipette 970 is connected to the suction end 90, the suction end 9811
It also serves to keep the fitting force between the suction end 981 and the pipette 97 constant by allowing it to move against the elasticity of the spring 108. In this way, the bellows pump 98 is removably arranged, and the bellows pump 981J can be removed by itself! ! - It is a mourning that has been made possible. Note that the tip of the suction end 98M is tapered to have a diameter #i1m toward the tip so that it can be easily fitted into the pipette 91.

Returning to FIG. 9, the bearing member lO2 is formed of a cylindrical body containing a collar S in the upper RAs, and is located at the ceiling 1 of the culture chamber 3.
It is fitted into a nine through hole drilled in i48, and is fixed to the ceiling 1148 by being prevented from being pulled out by the flange. The third bearing member 10m is equipped with a ball bearing machine near the upper and lower ports on the inner peripheral surface, and is fitted with the above-mentioned rotating sliding ring 101 made of a cylindrical body, so that the tube can rotate and slide freely. I support it. Nine, the output gear 104 has a wider meshing surface than the drive gear 103, so that even if the rotational sliding shaft 101 moves in the vertical direction, the connection between the output gear 1G4 and the drive gear 103 is always maintained. The engaged state is maintained. Further, the solenoid 11M1 and the motor 10s are housed in a housing 111, and the housing 111 or a board 11 integrated therewith are housed in a housing 111.
It is fixed at 11.

Note that, although not particularly shown, this dispensing device 11 includes:
Several position sensors are provided to automatically control the rotational position of the rotary arm 990 to a proper predetermined position. Further, as shown in number 41, at the position where the rotary arm 99 has rotated most clockwise,
The suction end of the bellows pump 9$ is 98m long, and the number of transfer wheels is 601.
It corresponds to the outer peripheral edge of the two mounting members 42. Hereinafter, the position of the mounting member 42 to which the suction end 98 of the bellows pump 98 corresponds, or the position of the mounting member 42
The position of the bellows pump 98 corresponding to this will be referred to as the dispensing position. 9. Transfer point 1iII at this dispensing position
At the outer position in the radial direction corresponding to the mounting member 42 of No. 6, there is disposed a lid-open milk position 78, which is arranged so as to correspond to the liquid supply position and the liquid waste position, and is the same as the companion one.

Needless to say, this lid opening/closing device 7B has the role of opening 114m from the Petri dish 4 during stirring or dispensing operations, and re-closing rJ*4m to the Petri dish 4 when these operations are completed.

As shown in FIGS. 1 and 2, the pipette supply wheel number 9s has its main body disposed near the rear right end of the machine room 47,
97 t It is designed to be supplied one by one.

As shown in the 101st KI, this pipette supply device 9i has a center S fixed to a rotation shaft 112, a plurality of pipette storage notches 1131 are formed at equal intervals on the outer periphery, and a nine-rotation disk 113. The above pipette storage notch 111
11 and engaged with nine pipettes 97,
A pipette supply lever 114 for dropping the pipette from the notch 1131, a solenoid 115 for retracting the pipette supply lever 114 to a non-supply position, and a screw for biasing the pipette supply lever 1141 toward the supply position. Shiba spring 116, and the rotating disk 113. It houses the pipette supply lever 114, etc., and a through hole 117m for pipette passage is formed in a part of the bottom wall so as to correspond to an opening (not shown) made in the ceiling 1i411 of the culture chamber 3. A housing 117 is formed in the housing 117, and an opening 118m corresponding to the through hole 117J1 is disposed so as to be slidable in an airtight manner on the lower surface of the bottom surface of the housing 117 by means of a guide means (not shown). A shutter plate 11g with a hole formed therein, a shutter drive lever 119 whose one arm end is connected to this shutter plate 118, and this Vya,
/ (-119 plunger 1 snow 1m at the other arm end)
The shutter drive solenoid 121 is connected to the shutter driving solenoid 121, and the provided companion pipe and toe receiver are the member 12 snow (see No. 111Q) and -
This pipette holder includes a shock absorbing coil spring 123 (see No. 1111Q) placed further below the member 122, and its main l! The department is made up of:

The pipette storage notch 113a is the pipette 97
Keyway-shaped vertical grooves are drilled symmetrically in the inner peripheral surface of the vertical hole, which has an inner diameter slightly larger than the outer diameter of the rotary disk 111, at positions corresponding to the inner and outer directions of the diameter of the rotating disk 111. It has a shape in which vertical grooves in the direction are communicated to the outer periphery of the disc 113, and a part of the upper surface of the disc 113 on the periphery of the notch 113!1 is provided with the upper end of the pipette 97. Shallow recesses 113b are formed at symmetrical positions to protrude and engage with the nine locking protrusions 90. This recess 15111b is symmetrically provided on the right side with respect to the longitudinal groove in the outer radial direction and on the left hook side with respect to the longitudinal groove in the inner radial direction, when viewed from the center of the rotating disk 113. . This recess @ 113b is the above-mentioned notch 1111K
The pipette 97 inserted from above and held in the storage position by engaging the protrusion 971 with the recess 113b serves to prevent the pipette 97 from moving inadvertently.

The pipette supply lever 114 is formed of an L-shaped plate, is swingably supported by a support shaft 124, and is attached to the support shaft 124, with one end attached to the lever 11.
4 and the other end to the stopper bin 125, respectively, by a torsion spring 116 having a leg-closing habit.
It has a habit of rotating clockwise around the support shaft 124. The rotation of the lever 1140 due to this habit is normally regulated by the other arm end of the lever 114 coming into contact with the tip of the plunger 115a of the solenoid 115. In this regulating position, the pipette engagement 111114m protruding from one side of the one arm end of the lever 114 is
It has been retracted from the locking position with the pipette 97 housed in the notch 113m. Furthermore, when the solenoid 115 is energized, the plunger 111>1 is pulled into the solenoid 115, so the lever 114 rotates clockwise around the support shaft 1240 due to the elasticity of the spring 116, and the side of the arm is turned into a stopper. The pipette retaining portion 114a is attached to the bottle 125 so as to move to a retaining position where it can engage with the protrusion 97a of the pipette 97.

The shutter drive lever 119 is swingably supported on a support shaft 126, and has nine elongated holes 119 bored at the end of the arm.
1 f Bin 12 planted on shutter plate 118? The shutter plate 1111 is connected to the shutter plate 1111 by being fitted into the shutter plate 1111. Further, it is connected to the solenoid 111 by having nine elongated holes 119b and 128 drilled at the end of the other arm, and fitted into nine pins 128 implanted in the plunger 121m. When the solenoid 121 is energized, the shutter drive lever 119 pulls the plunger 121M into the solenoid 121.
Rotate clockwise around the support shaft IH to release the shutter plate 1.
1g, the opening 110 is the housing 1170 through hole 11
It is designed to be slid to a position that coincides with 71.

And Ryokan 117 Otori.

118M match, the pipette 9 that had already been dropped into the notch 11al by the lever 114
7, inner hole 117m, 118m and ceiling 1148
It passes through the 0 through hole and falls into the culture chamber 8 yen. Thereafter, when the energization to the solenoid 121 is reduced, the plunger 1zta protrudes from the solenoid 121, and the shutter plate 118 automatically returns to the closed position.

The member 12g of the pipette holder has a gate IIvr that is medium or larger than the outer diameter of the pipette 91, as shown in jllllllQ.
It has a cylindrical body that is divided into two in the vertical direction, and the above dispensing position is half ill 1! ml is
The support shaft 122b is arranged so that it can be opened and closed freely (WIL) 9, and the support shaft 122b is arranged so that it can be opened and closed freely.
The circumference of 32b is rotated counterclockwise and stopped in contact with the other cow 1!1t. This pipette receiver 1 member 112 is arranged at a position where the pipette 91 falling from the housing 117 to the culture medium 8 can be guided through or through a resonant guide means. Insert it into the center hole without any
The upper end of the projection 9719 of the pipette 97 is held at its open peripheral edge Kfi&.

(9) The coil spring 1111 is wound to be thinner than the outer diameter of the pipette 91, and while falling, the pipette holder fits the lower end S of the pipette 97 into the member 128, and at this time The pipette holder absorbs the impact caused by the pipette 970 falling by increasing the winding slightly by 1 it, and the pipette 91t is engaged with the member 122 in a cushioning manner, thereby serving to prevent the pipette 97 from rebounding. The position of the pipette holder where the member 12m is placed is $
111! wAK As shown, the above bellows pump 98
When the dispensing device 11 rotates to the perosp/pipette holder position and lowers the sliding shaft 101, the suction end 911 of the pump 98
1, the pipette holder is fitted into the upper end opening of the pipette 97 which is received and positioned by the member 122, and the suction end sea
A pipette 97 is attached to the pipette. Then, after moving the rotary sliding shaft 101 upward, the same shaft 101 is rotated counterclockwise, and the bellows pump 9
8 toward the dispensing position, it is fitted into the suction end 18a, and the pipette holder 111 is moved by the pipette 97.
The half 91121 of the t is pushed and rotated around the support shaft 122b, and the pot and pipette 91 attached to the bellows pump 98 are taken out to the dispensing position.

Then, the half portion 1118 moves nt(9) around the support shaft 1!2b by the elasticity of an elastic means (not shown), and returns to half the ring 15Klil! The pipette receiver will return to its original state.

On the other hand, as shown in FIG. 1, the pipette detachment device 96 is provided in the culture i13, and as shown in 11g, the pipette detachment device 96 is fixed to the rotating shaft 111 and the rotating wheel 1m1, and is connected to the nine cam lever 13s. , one end of which is engaged with the cam lever 112, and a compressible coil spring 133 that gives the lever 132 the habit of rotating clockwise about the rotation axis 131; and the cam lever 132 due to the elasticity of the coil spring 133. A stopper bottle 134 (see FIG. 12) restricts the rotation of the pipette to a predetermined pipette removal position.

The cam lever 132 has a flange 598 of the bellows pump 98 at the end of the nine arms that extend out from the rotating shaft 131.
A pair of 11111 forces are applied between C and pipette 91 to separate them.
After this separation is completed, the bellows boning 19 is attached to the inner ridge.
A round notch 132c is formed through which the suction end SSa of 0 passes. The above lI cam portions 13i1, 132
b is formed so as not to obstruct the movement of the suction end 981 and to extend in the radial direction again after being raised 9, and one cam @1sea has the stopper bin 134 A ninth l111112d that abuts is integrally formed. Further, the notch IHc is formed in the vertical direction of the double-edged arm portions 1328 and 132b with 1% IK, and when the cam lever 132 engages with the stopper pin 184 and is in the nine-sided position, the bellows pump 98's suction @ SSa's rotation locus corresponds to KN. The rotation shaft 111 is configured to rotate counterclockwise at a certain angle against the elasticity of a spring 1330 by a rotary reel maid or the like (not shown).
It is designed to evacuate the bellows pump from the path of movement of $.

The pipette separation device I6 thus llllt is
When the dispensing device 11 is rounding up the used pipette (97), move the pipette holder from the dispensing position to the o-x position.
When the Honda 98 is moved around, the cam lever 132 is in the pipette release position where it engages the stopper bottle 184K. Therefore, the bellows pump 98 is the same as the pipette 97tfi.
When the cam lever 112 moves to the tip position (9) in the attached state, 1lffi force 5Is 1328. INZb
The suction end 98 of the pump 98 is inserted between them, the bell @ 91C comes into contact with the upper slope of the cam @ 132m, 189, and the upper end surface of the pipette 97 is brought into contact with the cam 91321, 131.
It comes into contact with the lower body of 1b. Then, as the bellows pump 98 rotates further based on this foreshadowing, Ill @
It is pushed up by the upper slopes of the esc five cams 11111 and 131b, and the space between the collar 980 and the pipette 97 is pushed wider.

For this reason, the pipette 97 attached to the suction end e-1
The pipette 97 comes off from the suction end 11811 and falls downward due to its own weight, and the pipette 97 is removed.

This fallen pipette 97 passes through a pipette guide member (not shown) disposed below and culture! [3] The pipettes are introduced into a 9-pipette storage tank (not shown) and stored. The pipette 97 stored in this way is
It was later collected and cleaned.

After sterilization, it is reused. In addition, between the pipette storage tank and the culture ils, there is a dustproof and antibacterial filter made of a round, silicone/rubber thin film filter, etc. to prevent the culture chamber from being contaminated by ventilation between the two. It is provided. Please note that the bellows pump 98 is a used pipette)
97t - After detaching, the pipette holder continues to rotate using the lever and moves once to the position, and then, by reversing the direction of rotation of the rotating and sliding shaft 101, it moves back to the dispensing position again and carries out the next pipette. The field remains in a waiting state until the installation begins.

Next, in order to move the dispensing device 11 back to the bellows pump 98 dispensing position with the new pipette 7. The bellows pump 98 is rotated, and the cam lever 132 is rotated against the elasticity of the spring 133, so that the bellows pump 98 is retracted from the rotation locus. This round, bellows pump 98
without hitting the cam lever 132) 9
7 With the tfi attached, the cam lever 132 moves back to the dispensing position, and then the rotational force of the rotating wheel 131 is removed (by the elasticity of the spring 13B), and the cam lever 132 collides with the stopper via 13.4. Return to pipette-arm position.

The petri dish supply device 12 has M I II and a second
As shown in the figure, a main body part is disposed in the rear part 11 of the machine room 47, and a chute 131 for guiding a dish dish is formed so as to project out from this main body part into three circles of the culture chamber. There is. The main body of the petri dish feeding device 1 is a first
．． As shown in 11E, a round petri dish storage part 114 in which sterilized petri dishes 4 are stacked diagonally and stored;
This petri dish storage 51311 has a lower end sK,
A housing that surrounds the circular transfer @ 137 that takes out petri dishes 4 one by one from the same 5136 and sends them out to the shooter 1s5, and this circular transfer @ 1ift handle, and whose bottom surface also serves as the bottom surface of the circular transfer @ 187. 18111
and a round wheel 13G provided at the center of the circular transmission @ 187, st-m determined based on this rotating shaft 18,
A rotating arm 141 whose free end hits the petri dish 4 and pushes it; and a rotary arm 141 whose free end hits the petri dish 4 to push it; The hole 117 and the bottom surface of the housing 13 are arranged so as to be able to swing in an airtight manner by a guide means (not shown), and the hole 142m corresponding to the hole 187 is bored. A shutter plate 142 is provided, one arm is connected to this shutter plate 142 to drive the nine shutters, and a shutter driving lens 144 is connected to the shutter driving lever 1480 and a plunger 144 is connected to the other arm end. , its main parts are comprised.

The petri dish guide tray 131S has a horizontally long rectangular cross-sectional shape that allows the petri dish 4 to pass through, and is composed of nine tubes. through machine 1ii
The petri dish feed opening 18i1 at the upper end of the opening 18i1 is bored into the bottom surface of the housing 1811, and the shutter plate 14m slides into the nine opening 1371. They are facing each other via mt for as long as possible. As shown in FIG. 4, the petri dish delivery opening 13Sb at the lower end corresponds to the upper part of one loading member 42 in the transfer device 6, and the opening 13Sb is connected to the opening 13Sb through the shooter X5St. The petri dish 4 falls onto the mounting member 42 due to its own weight, and the same member 411
It is set to be placed on. Below, this narrow mouth 135
The position of the mounting member 42 directly below b will be referred to as the petri dish supply position.

The petri dish storage 5136 is designed to store petri dishes 4 diagonally stacked as described above.
13@Yoshi circular transfer@137 When the petri dish 4 sent to 9 is pushed and moved by the rotating arm 141 as shown in FIG. 13, the petri dish 4 stacked on top Due to its own weight, Petri dish 4 at the bottom is i.
It is configured to be dynamically sent to the circular transfer section 137 yen. The rotating arm 141 has a circular free end.
A rounded claw portion 141a is formed on one side of the peripheral wall of the container 187 in the direction of rotation for catching the petri dish 4 sent out from the storage 5136. This rotating arm 141 is fed out from a rotating shaft 136 rotated counterclockwise by a motor or the like (not shown), hooks the nine Petri dishes 4, and circular transfers 1ii.
While pushing along the circumferential wall of 1137, Petri dish 4 is once moved to the waiting position in front of the open hole 137 field and stopped. Then, when it is time to feed the petri dish, the rotation starts again in the counterclockwise direction, and the opening 14211 of the shutter plate 14g moves to correspond to the opening 1!71 and the same hole 1ava in synchronization with the rotation. familiar with, shar
The power is supplied to the -V4t and -tar 13I.

The shutter drive lever 14m has a support shaft 14g1K.
It is pivotally supported so that it can swing freely, and has nine elongated holes 1 bored at the end of the arm.
431 f Bin 180 planted on shutter plate 141
It is connected to the same plate 142 by being fitted into it.

In addition, a round pin 146tIIIC is inserted into the nine long holes 143b and the plunger 144m is drilled in the other arm,
It is connected to the solenoid 144. When the solenoid 144 is energized, the shutter drive lever 148 moves from the solenoid 144 to the plunger 14411.
protrudes and rotates counterclockwise around the support shaft 141 from this k to slide the shutter plate opening 2 to a position where the seventh opening 141m matches the opening 137m of the housing 188. It has become. Both holes 117m, 141m
The petri dish 4, which has been pushed and moved by the rotating arm 141, is in good condition and matches the sM hole 137m.

Needless to say, the shutter plate 142 falls through the shutter 131s circle through 1428f.
When the energization to the solenoid 144 is released, the same solenoid 1
The plunger 1441 is retracted into the shutter 44 and automatically moves back to the shutter closed position.

According to the nine-petri dish supply device 12 configured like a petri dish,
Every time the rotating arm 141 rotates 1, one Petri dish 4 stacked in the storage section 136 is transferred to the circular transfer section 1!
7, and the openings 137m and 1421
The petri dishes 4 are delivered one by one to the shooter 135 through the feeder. Therefore, the petri dishes 4 are automatically supplied one by one onto the mounting member 42 of the transfer unit 11t6.

As shown in FIGS. 1 and 2, the observation device 14 has an eyepiece portion protruding outward from the front S* of the right side wall of the housing 2, and an objective portion, f, and a source portion for incubation. It is housed in a circle and is made of a mirror, and is placed above the optical aSt objective section, and is formed in a so-called inverted shape. The eyepiece section is formed in the same way as a normal microscope, whereas the objective section requires moving the objective lens 147 from the outside to a rounded focusing point. The drive mechanism is electrically driven. That is, as shown in detail in FIG. 1s, the driving mechanism for the objective lens 147 is attached to the upper circumferential surface of the objective lens 147, and a helicoid is attached to the outer circumferential surface of the outward flange formed on the upper l11i1sIF. A base shaft sliding tube 148 in which a male thread 148a is carved and a guiding elongated hole 148b in the optical axis direction is drilled in a part of the lower outer circumferential direction, and this base shaft sliding tube 148
1, the lower end mt
51, and the outer circumferential surface of this bearing cylinder 151 is generally housed in a nine-concave hole, and the same cylinder tslK11 is formed, and its tip S is inserted into the guide slot 148b of the original shaft sliding cylinder 148.
Fitted into a circle, the friend guide bin 15m and 1 above-mentioned bearing tube 151
A lower circumferential surface is rotatably fitted to the outer periphery of the optical axis sliding tube 148 via a ball bearing mechanism 1531, and a helicoid plow screw 154a threaded on the upper inner circumferential surface is connected to the optical axis sliding tube 148.
A rotary cylinder 154 that is screwed onto the helicoid male thread 148a and has a gear 154b engraved on the outer circumferential surface of the outward facing flange of the upper end, and an output gear 1s1 meshed with the gear 154b of the rotary cylinder 154. The main part of the output gear 155i is composed of a nine-pin gear 156 attached to the output shaft.

In addition, on one side circumferential surface of the lens barrel of the objective lens 147, there is an index 1 made of a light absorber that indicates the operating range of the lens 147.
4G is coated, and an optical sensor 9-150 is provided so as to face the indicator 140.

This optical sensor 150 completely detects whether or not there is an index 14G at a position facing the sensor 150, and
In pigeon houses where the objective lens 147 is not detected, the objective lens 147 is outside the operating range, so a buzzer (not shown) generates a warning sound and the motor 1 for driving the objective lens is activated. ! 6
It is designed to stop the rotation of the machine. First, the motor 156 is connected to a switch member (
(not shown), the objective lens 147 can be rotated in either the forward or reverse direction.
The optical axis is arranged so as to be located directly below one of the mounting members 42, and as shown in FIG. 4, the optical axis almost coincides with the central axis of the seven mounting members 42. . Hereinafter, the position of the mounting member 42 directly above the objective lens 147 will be referred to as the am position. Therefore, the culture cells in the Petri dish 4 that are placed on the mounting member 48 at the observation position,
It is designed so that the lower surface can be clearly observed through the objective lens 147.

In addition, in FIGS. 1 and 2, the symbol 1srht*@vnx1(.

Also, in Fig. 14, the symbol 18g is the objective lens! 47 and a relay optical system optically connected to the eyepiece lens 157. Furthermore, although not particularly shown in the drawings, since the culture medium is kept under pressure at a high temperature, the objective part of the observation device opening is formed into a moisture-proof structure that makes extensive use of bonding of parts, backing, etc. There is.

With the objective section of the companion observation device 14 configured in this way,
To perform the focusing of the objective lens 147, external f)
Operate x switch iB# to rotate motor 1t@1 clockwise and counterclockwise K11. Then, output gear 1
55, the rotation tube 154 is rotated counterclockwise or clockwise, and the bottle 152 and the elongated hole 1411b move forward and backward in the optical axis direction. Therefore, the eyepiece 157 color transparent 11
Body 4F) While observing the culture m1lilot in Petri dish 4, just operate the switch member to stop the rotation of the objective lens 156 when it is in focus. 147
If the optical sensor 15 is out of the operating tolerance range, the optical sensor 15
Since the index 140 is no longer detected, no warning is given or the motor 156 is automatically stopped. Therefore, the objective surface of the objective lens 1470 may collide with the petri dish, etc., and the petri dish may be damaged or the lens may be scratched.
．． ．． There is no risk of it happening.

On the other hand, as shown in FIG. 1, the illumination light source section of the observation device 14 consists of a light source lamp 161 and a condenser lens 162 housed in a moisture-proof outer tube ls9.
811iltxao, and the ceiling 114
The outer mantle cylinder 18G is fitted into the through hole installed in
It is fixed to and placed so as to hang down within the culture ii3. It goes without saying that the optical axis of this illumination device 1600 coincides with the optical axis of the objective lens 1470.
A moisture-proof glass plate 164 is fitted through an annular holding member 166 and an O-ring-shaped backing member IH, so that there is no problem in emitting illumination light. Furthermore, in this illumination device 160, filters, etc., which are not particularly shown in the drawings, can be replaced without the machine 1147, and the illumination light can be adjusted without having to clean the inside of the culture chamber 3. It's starting to get cloudy.

The observation device 14 has a KIR Schilling gold in the illumination device 1, and a position S*t-1 in the observation optical system, so that it can also be used as a phase contrast mirror. In addition, li@ilK is equipped with an optical system for photographing, and it is possible to take photographs of cultured cells in the petri dish 4 without taking the shear 174 out of the culture chamber 3. It looks like this.

By the way, although not shown in particular in the culture room 3,
A heating and humidifying device that heats and humidifies the inside of the culture room 3, a round preliminary heat-insulating device that keeps the cultured tame circle at a predetermined temperature, and a l1lll of the culture room 3! I medium pH 7 appropriate value (approximately 1.
2) A carbon dioxide gas/air supply device to maintain the temperature, an ultraviolet sterilization device to sterilize the culture room 3, and an air blower device to purify the air in the culture room 3 are attached.

The above heating/humidifying device consists of a water tank that stores water for humidification, a heater that heats the water tubes in the water tank, a temperature sensor that detects the temperature sound of the water in the water tank, and a temperature sensor based on the output of the temperature layer sensor. A temperature control device that controls the power supply to the heater to keep the water temperature constant (for example, 37 degrees Celsius), a blower that blows warm air from the aquarium into the restaurant, and a heating
It consists of a silver volume and a ventilation pipe that connects the culture chamber 3.

This is heated by a heater to keep it at exactly 37°C (within ±01°C), and then cultured using a humid warm air pipe blower above the tank. At the same time, air from the culture chamber #13 is circulated into the water tank to warm and humidify the culture chamber.

The preliminary heat retention device is composed of nine heaters that are evenly distributed around the circumference or outer circumferential surface of the culture@S, and a temperature regulator that detects the temperature of the heater and controls it. , the culture medium is heated to a temperature [(e.g., age) that is closer to the target temperature (e.g., 87° C.) than the surroundings and lower than this. By pre-warming the culture 1iat-i in this way, even if the air flow inside the culture chamber 3 due to the heating/heating device described above is uneven, the temperature will vary greatly depending on the location. Troubleshooting is the key to troubleshooting.

The carbon dioxide gas/air supply device is used to keep the carbon dioxide concentration in the gas atmosphere in the culture chamber 3 constant and maintain the pHt of the culture solution in the petri dish 4, which is a single cell culture medium, at an appropriate value (approximately 7.2). It consists of a box containing carbon dioxide, an air pump that takes in air from the outside, a gas flow needle that controls the gas flow rate, and an electromagnetic valve that controls the gas flow rate. It consists of a connecting air pipe.

'This device combines the carbon dioxide gas taken out from the bottle/vehicle at an appropriate pressure and the air taken from the outside by an air pump through the air intake port in an appropriate ratio (carbon dioxide gas 591I). , air (90°C) and fed into the culture chamber.

The above-mentioned sterilization device 1iI is designed to turn on an ultraviolet lamp and sterilize germs with the irradiated ultraviolet light when the inside of the culture chamber 3 is contaminated or becomes contaminated. To9 consists of an ultraviolet lamp and a ninth operation switch that controls the blinking of the lamp. By the way, the method of sterilization using ultraviolet rays has the disadvantage that areas that are not irradiated with ultraviolet rays are not irradiated.
, In the case of this automatic culture 111tl, the culture room is 30 yen I1
The surfaces of the equipment installed in the 3rd circle are made of extremely polished stainless steel, and due to their mirror effect, the ultraviolet rays are reflected and irradiated to the back of the equipment, allowing for a wider range of coverage. It has been devised so that it can be easily destroyed.

The above-mentioned clean air blowing device is composed of high-pressure blowing powder and a high-performance filter.
This device, which dries and cleans the inside of the culture gS by blowing in 9 air (for example, 11$Oj/Ith or more), is operated in an oscillating manner when cells are not being cultured. It is used for caulking and purifying the inside.

Furthermore, the front jar of the 3 yen culture room is made of glass gkp so that the inside of the culture room 3 m3 can be directly observed from the outside. However, an outer door that can be opened and closed is provided on the dew condensation t-ti to cover the front side. This outer door is equipped with a heater and a temperature sensor.
The same layer is 37℃~40'CI which is slightly higher than culture 113 yen!
It is heated to a certain degree. As a result of this heating, the temperature of the glass chamber on the circular side decreases.

As shown in No. 111, the control device 18 includes an arithmetic processing device such as a large mital combination, an arithmetic processing device, etc., which is incorporated into a display and operation panel 167 provided on the front left side of the housing 2. It consists of input/output devices and a power supply.

The display/operation panel 167 has a liquid crystal display panel on its front surface 11 for displaying the temperature inside the culture chamber 3. A display 118 is provided, and a temperature adjustment knob 169 for adjusting the temperature of the culture 11H is provided in front and below it. Furthermore, below it, a plurality of rounded various operating members 171 for controlling the operating tube of the automatic culture device 10 are arranged in a row.

The arithmetic unit mi device has a petri dish 4 loading/unloading program, a waste liquid program, and a liquid supply program I-gram.

The stirring program and the dispensing program are programmed with information such as grams, and the automatic culture device 1 is configured to control the one-shot operations required for automatic culture based on these programs. There is. That is, after reading the outputs of the solenoid in the motor built into the nine various types of silver as described above, the automatic culture device 1 was activated. IIjA automatically
It is designed to carry out a series of operations required in subculture of IIK.

As described above, the culture container transfer device 6 of the present invention and the automatic culture device 1 installed therein are constructed.

Next, regarding the operation of the culture container transfer device of the present invention,
The operation of the automatic culture device 1 will be explained with reference to the flowchart shown in 1st @IEI.

First, operate the power switch (!@I not shown) of the automatic culture device 1 to put the device 1 into operation.
The humidifier, pre-warming device, carbon dioxide gas/air supply device, etc. are activated, and the inside of the culture chamber 3 is suitable for the cell culture 111 m1k-ji
INOIIIIL (Il&sy℃, im degree tools member acid gas concentration! 11) K is set automatically.

Next, when the culture command switch (not shown) of the automatic culture device 1 is operated, the nine mold loading/unloading devices shown in No. 1 m11 are activated, and each belt conveyor 2s, 2.

39 operates in the direction of carrying in the petri dish 4, and the process of carrying in the petri dish 4 is started. That is, if nine petri dishes containing Sat cells to be cultured (4t) are placed on the sheath 32 and fed into the loading/unloading device 5, the same petri dishes 4 are transferred to the belt conveyor 2G.
, ! 11 and 23, and transferred to culture room 3. During this transportation, Peltoco/Pair 3
s is linked to the opening of the shutter 24, and the supporting shaft = 3! The l horse 9 is rotated by a certain angle in a clockwise direction and is fitted into the notch 42b of the mounting material 42 at the loading/unloading position KTo. Therefore, the petri dish 4 is automatically set on the device member 43 by the conveying force of the belt conveyor 33. The setting of the petri dish 4 on the mounting member 42 is detected by a carry-in/unload detection sensor (not shown), and the operation of the carry-in/unload device S is stopped based on the output of this sensor. Then, in conjunction with the closing of the shutter 240, the belt conveyor 23 rotates counterclockwise around the circumference 9t of the support shaft 2I, returns to the flat position tilted from the horizontal position, is withdrawn from the notch 4sb of the mounting member 42.

Following the stoppage of the loading/unloading device S, the transfer device 6 of the present invention shown in FIG. 4 is rotated clockwise (in FIG. Here, the operation unit uses the observation device port to confirm that the nine Petri dishes 4 containing the cells to be cultured are set in the predetermined position of the culture i13.Then, after this is confirmed, the culture Operate the continuation command switch (not shown).Then, the process of draining the culture liquid starts.In this process, first, the transfer device automatically rotates clockwise, and the Petri dish 4 is the lid opening mi device 7 corresponding to the zero-order Kli liquid position moved from the observation position to the waste liquid position.
0 is activated and 4 m of petri dish 4 is released. Subsequently, the solenoid 64 of the device 7 (see Figure 1) is actuated for the waste 11, and the tip s8 is supplied to the tip insertion/removal tube 66. Then, the gauntlet hs is activated and the main body is moved) 1
18, the drain pipe 51 is gradually lowered.

During the descent of the drain pipe s1, the chip insertion/removal 1
Fit the 66 yen tip 58t into the tip m, pass through the tip insertion/removal tube 66, enter the tip of the tip 5m into the petri dish 4 yen in the waste liquid position, and push and move the petri dish 4t to tilt it slightly. stops at the 9th position. Next, a suction pump (not shown) is operated for a sufficient period of time to suction and discharge the culture solution from the petri dish (4 yen) through the P liquid pipe 51 yen to the lower end opening of the chip 58.Next, the motor 5s is activated. The drain pipe s1 is rotated in the opposite direction to the first rotation, and as the drive belt 53 moves, the drain pipe s1 is moved upward. Drain pipe 51
When the petri dish number is raised, the petri dish number returns to the horizontal position, and during its rise, the drain pipe 51 abuts the upper surface of the chip 5s against the lower gIAlli of the chip insertion/removal tube 66,
The chip s8 is dropped from its tip. Then, when the motor 5s moves back to the desired position, the rotation of the motor 5s is stopped and the drain pipe 51 returns to the normal position. The used chip 58 that has fallen off from the tip of the drain pipe 51 is taken out of the culture i13 through a guide means (not shown) and placed in the housing 2.
The 9-chip storage cypress (shown in the figure) is disposed near the bottom of the 0 and the lid opening/closing device 18 is operated again, and the rice 4 of the petri dish 4 is closed.

Subsequently, a cleaning liquid injection process is started. In this process, first, the transfer device 6 is rotated in the counterclockwise direction (in FIG. 4), and the petri dish 4 is moved from the liquid waste position to the liquid supply position. Next, the lid opening/closing device 18 corresponding to the liquid supply position is activated, and a petri dish number of 4m is released. Next, the 8th
The nine roller pumps 74b shown in the figure are operated, and the cleaning liquid is supplied from the storage container 72b through the roller pump 14b, the warmer 75b, and the liquid supply tube 76b Yr to a fixed amount (e.g., scc) to 4 yen. be done. Then, the lid opening/closing device 78 is operated again, and the lid 4 of the petri dish 4 is closed. The washing process of kernels
Wash away the old culture solution that adhered to 11, and then remove the remaining 114.
6. In science and engineering equipment, what is done to enable enzymes to function effectively is the same as described above.

Next, a post-cleaning liquid waste process is performed to discharge the cleaning liquid injected into the petri dish 4 in the cleaning liquid injection process to the outside of the petri dish 4. In this step, first, the transfer device [6 shown in FIG. 4] is rotated clockwise, and the Petri dish 4 is moved from the liquid supply position to the liquid waste position. And from now on,
The cleaning liquid is discharged from the petri dish 4 in exactly the same manner as the culture medium** liquid waste process described above.

Then, after the washing layer draining process is completed and the animal is dead, the enzyme solution injection process is started. In this one food, first, the transfer device 6 shown in Fig. 4 is rotated counterclockwise, and the Petri dish 4 is
is moved from the liquid waste position to the liquid supply position, the lid opening/closing device 78 is operated, and the petri dish 40 and 41 are opened.
The nine roller pumps 74G shown in Figure K2 are operated, and a certain amount (for example, 3 cc) of the enzyme solution is pumped into the petri dish 4 from the storage container 72C through the roller pump 74C and the warmer 75C9. Supplied. Subsequently, the lid opening/closing device 78 is operated again to close 44 m of the petri dish 4, and the petri dish 4 is left stationary on the mounting member 42 for about one minute. This standing allows the enzyme (for example, trypsin) in the enzyme solution to sufficiently act on the cultured cells implanted on the surface of the petri dish 4, thereby ensuring that the cultured cells are released from the bottom surface of the petri dish 4, which is the growth surface. rounding is done.

Next, one injected amount of the enzyme solution is injected into four Petri dishes, and a process of draining nine enzyme solutions is carried out. This l111 [process is carried out in the same way as the cleaning liquid I & Tanko Yang and Yu (I will omit the popular theory).After the waste liquid process of this enzyme liquid is completed, Kuyare 4 will be approximately The petri dish 4 is allowed to stand still for 10 minutes on the plate @@4'A.
II4 This is carried out in order to promote the release of the turtle and the growth surface, as well as to weaken the bonds between each cell and promote the monomerization of the cultured cells.

When the petri dish 4 is left still, a cell detachment step is next performed. In this step, first, the transfer device 6 shown in FIG. 4 is rotated clockwise K111, and the petri dish 4 is moved from the waste liquid position to the stripping position. Next, the 81st
The solenoid 91 of the peeling device 90 is intermittently energized for 1 minute, etc., and the hitting member 9s continuously hits the Petri dish 4 or 4tii4a on the surface of the Petri dish 4 to apply lateral vibration to the Petri dish 4. When this vibration is applied, the cells to be cultured which are free from the bottom surface of the petri dish 4 are subjected to a lateral shearing force due to their own inertia, and are reliably peeled off from the bottom surface based on the shearing force.

Next, after the above detachment step is completed, the culture solution injection factory is first destroyed in order to separate the detached cells into individual cells. In this process, first, the transfer device 6 shown in FIG. , 64m of Petri dish 4 was opened, and then No. 2 I
The roller pump 74m shown in 11 is activated. As a result, the roller pump 74a, from the storage capacity of 5721 yen,
Heating 1 ust, lI! 1 liquid chi.

- Pour one volume of culture solution (e.g. 3 c
c) Only 4 yen of petri dish is supplied. Then, the lid opening/closing device 7 is operated, and the Ii4 chamber of the petri dish 4 is closed.

Subsequently, a step of stirring the culture solution is performed. In this step, first, the transfer device 6 shown in 4aA is moved to the temporal position. On the other hand, at the same time, the nine pipette supply silver amounts aS shown in FIGS. 10 to 12 are activated, and the pipette 97 is supplied to the member 13 and held there. Then, the rotary sliding wheel 101 of the dispensing device 11 (see Figures 11.11 and 12) is rotated counterclockwise, and the bellows pump 9
8 is moved from the dispensing position to the pipette receiver position. Subsequently, the rotating sliding shaft 101$ is lowered, and the bellows pump 9
The suction end 98m of the pipette 97 is fitted into one opening of the pipette 97, and the pipette 97 is attached to the bellows pump 98. Then, when the cam lever 132 is rotated counterclockwise about the rotation axis 131t and temporarily evacuated from the locked position with the nine pipettes, the rotation and sliding axis 101 is moved upward. Then, the bellows pump 98 to which the pipette 97 is mounted is rotated in the direction of the hour hand and moved back from the pipette receiver position to the dispensing position.

Next, the lid opening/closing device 7S corresponding to the dispensing position is activated,
Petri dish 4, 948, is opened. Subsequently, the rotating and sliding shaft 101 is lowered again, and the tip of the pipette 97 is fitted into the Petri dish 4tl' at the dispensing position.
The pressure is F, and the tilt is in good condition, and the descent is stopped.Next, the bellows pump 98 is activated to suction, and the pipette 9 pumps a certain amount (for example, $ CC) of the culture solution in the petri dish 4.
7. When the rounded and peeled cultured lipids are separated from each other in the Petri dish 4 circles, when they pass through the thin suction port of the pipette 91, they are separated from each other and further made into individual pieces, and are transferred together with the culture solution into the pipette 97. drawn. After suctioning this culture solution, the sliding wheel 101 is raised again, the bellows pump 9s is operated in the discharge direction with the pipette 97 at a high position KTo, and the culture solution in the pipette 97 yen is sampled *
Together with the S cells, it is discharged again into a 4-circle petri dish. Therefore, the cultured cells are further isolated. Pipette 97 above
The suction and discharge of the culture solution is repeated 10 times 9 times, and the culture solution is sufficiently stirred to completely separate the cultured cells into individual cells.

Following the above stirring process of 1 mK, a culture solution dispensing process is performed. First, the rotary and sliding shaft 101 is lowered, and the culture solution containing the single cultured cells in the petri dish 4 is pumped into a certain amount (
For example, s 3CC) 711! and aspirated into the pipette 97. Subsequently, the lid M@ 78 is operated and 114 m of the petri dish 4 is closed.

At the same time, the petri dish feeding device 12 shown in FIG. 13 is activated, and the mounting member 4 at the petri dish feeding position is
A new Petri dish number 4 is supplied and set on the Petri dish number 2 (hereinafter, this new Petri dish number 4 is referred to as the first Petri dish number).Next, the nine transfer device shown at 11411 places the Petri dish [1701 The Petri dish feeding device 13 is operated again, and another new Petri dish 4 is placed on the cutting member 42 at the Petri dish feeding position.
is supplied and set (hereinafter, this new petri dish 4t).

Next, the transfer device 6 is rotated clockwise. , the first petri dish 4 is moved to the dispensing position. Then, the quantity opening/closing device 11 is operated, and the petri dish 4 of 1I11 is
The lid 4a is opened, and then the bellows pump gS is discharged, and half of the culture solution in the pipette 97 containing the single cultured cellulose III (for example, 1 cc) is discharged.
4 yen is discharged into the first petri dish. Then, the #i opening/closing valve S is operated again, and the amount 4a of the first petri dish 4 is closed. Subsequently, the transfer device 6 is rotated clockwise by one Petri dish placement 19137, and the Petri dish 4 at @2 is moved to the dispensing position. 4 open on 9!
The #1% device 78 is activated and the lid 4 of the petri dish 4 of w&z is activated.
a is opened, the bellows pump 98 performs the discharge operation again, and the remaining half of the culture solution containing the singulated cultured cells in the pipette 97 is discharged into the second petri dish 4. Then, the l1llll'l emplacement 7S is operated again, and the fi4m of the second petri dish 4 is closed.

In this way, the culture solution placed in the first and second Petri dishes 4 is only half of the predetermined amount, and as it is, there is not enough light to culture the cells, so the amount of culture solution is insufficient. Next, a culture solution refilling step for replenishing the first and second petri dishes 4 is performed, and the tray 6 is rotated counterclockwise and moved from the petri dish 4 dispensing position of tic 2 to the liquid supply position. Next, after the lid opening/closing device 78 is activated to open the lid 4a of the second petri dish 4, the row 2 pump 74a shown in FIG.
The culture solution is pumped through the roller pump 74m, the warmer 75, and the liquid supply tube 741a from
C) is supplied into the eighth petri dish 4. and,
The lid opening/closing device 78 is operated again to close the lid 4a of the second petri dish 4. Next, transfer crack 1 shown in Figure 84
lll16 moves counterclockwise again to the petri dish Ikuo 37.
The first Petri dish number is moved to the liquid supply position. After this, the lid N closing device 78 is activated and the first
Petri dish 4 1i411 is opened, and the roller pump shown in FIG. 4 petri dish is activated and the first petri dish 4
4 shortages (for example @cc) of culture medium are supplied in the container. Then, the lid opening/closing device 18 is operated, and 114 m of the first petri dish 4 is closed.

After the compensator IK of the culture solution is completed, the aliquot device 11
(Chapter e, tl, tsllllllll) K@ Karasugata 1m of used pipette 97 left unattended
will be carried out. That is, the rotating sliding wheel 101 is rotated once in the counterclockwise direction, and the bellows pump $ is rotated toward the pipette receiver position. Ninth, the bellows pump 9 moves to the tip position of the cam lever 1811, which has moved back to the pipette release position where it engages with the stopper bin 114K.
Suction end 5sati+ui force A part 13m1, 1811
) In addition to inserting it in between, Taku@e Hatake C and Pipette 7
Between both cams 910 and 1llb, insert the tip of the 910 and insert the 9ft tip of the 90mm.
Therefore, the nine pipettes fall off due to their own weight (p, suction end, and S), and through a mechanism (not shown), the culture i1i
9 taken outside, 30all cases! The 0-rotation sliding shaft 101 stored in a pipette storage tank (not shown) disposed in the pipette 1wn rotates the bep pump 9 to the bivet holder position, then it is reversed and rotated again. The bellows pump 98 is moved back to the dispensing position and then stopped.

Next, after taking out the culture 1Ilt in which the individualized cultured cells meet, a used old petri dish and an automatic culture device 1 are used.
A round Petri dish (4) is placed outside. In this process, first, the round plate feeding device 411 is rotated, and the old petri dish 4 is moved to the loading/unloading position.Next, the loading/unloading device 6 is operated, and each belt conveyor ,8・.

The machine 9 is operated in the unloading direction, and unloading of the petri dish 4 is started. That is, when the shutter 34 is opened, the belt conveyor 23 is rotated in the direction of the hour hand around the support shaft ss in conjunction with this, and the notch 4 of the mounting member 4s is moved from the round end to the loading/unloading position KTo. The old petri dish 4, which has been inserted into the container b and placed on the snow, is moved onto the co/bear zs by the action of the engaging claw Hb and the conveying force of the conveyor 3s. It will be posted. Then, it is conveyed for 1 second on the PeltoCo/Bear 3s, 2, 89, and sent onto the tray ss by the conveying force of the conveyor 29. Therefore, it is better to manually pick up the petri dish 4e) from the top of the race s3 and shake the ξ angle.

Then, as described above, the cells of the nine-generation culture system created in the first and second Petri dishes 4 are started to be cultured. This culture process IM is performed by leaving the first and first petri dishes 4 still for a long period of time (for example, S days) in the culture chamber 3 maintained at a constant pressure. When the petri dish 4 is left still, the cells to be cultured which are singly suspended in the second petri dish 4 settle in the culture solution and settle on the bottom surface 11 of the petri dish 4. Using this as a growth surface, cell division 1 & IK
Since the culture II contains an abundant source of nutrients and is maintained at the optimum temperature and pH for proliferation, the cultured cells will proliferate reliably.

If the specified patent required for proliferation has passed and the period has passed, operate the round plate transfer command switch (II not shown) provided with the automatic culture storage IK to move the transfer device t-a@, and transfer the first petri dish 4t.
Move it to the lll11 position. And the observation device 14
As a result, the proliferation of cells in 4 circles of Petri dish 1 is mtm*. In addition, in the same manner, the growth state of the mesocystis in the petri dish 4 of 1I2 was determined to be 1 mm.

Observation of the cells in the first and second Petri dishes 4 reveals that the proliferation state is not sufficient and if the proliferation is to be continued, the first and second Petri dishes 4 are left as they are. Maku,
When a predetermined growth state is obtained and it is desired to create a next-generation culture system using the proliferated cells as cultured cells and conduct a first-generation culture, the culture continuation command switch (lcl not shown) is operated.

Then, as shown in FIG. 16, the first and third Petri dishes 4 are again subjected to the process from about 5niI of the culture solution to the disposal of the Petri dishes 4, and the next generation am culture system is placed in the trap. A third generation culture of #I cells is created.

Then, by observing the cells in the first and second petri dishes 4, if it is determined that no further culturing is necessary, a step of carrying out the first and second petri dishes 4 is performed. In this unloading process, the first and second petri dishes 4 are transferred to the image loading/unloading position, and the loading/unloading device S This is done by installing two actuating cylinders. The first and second petri dishes 4 that have been carried out are sent out one after another onto the tray 32, so these petri dishes 4 are picked up manually and the purpose of the amt value grown in the petri dish 4 yen, for example, to Just use it.

As described above, according to the present invention, the culture medium circle can be cultured without touching it from the outside! Since the culture vessel can be moved to a predetermined position, there is no risk of damaging the atmosphere inside the culture vehicle compared to the conventional case of manually moving the culture vessel.Therefore, there is no risk of adverse effects on cultured cells due to changes in environmental conditions In addition to being able to remove the particles, accidents such as contamination with various germs can be effectively prevented. Therefore, since standardized culture can be performed under negative conditions, uniform cultured cells can be stably obtained, and the reliability of experiments using cultured cells is significantly improved.

Furthermore, since there is little risk of cultured cells leaking out of the culture room, there is less risk of biohazard, and culture can be performed with a high degree of safety.

Therefore, it is possible to provide a culture container which is extremely convenient to use and which eliminates the drawbacks of the conventional method.

[Brief explanation of drawings]

111th! 1 is a front am cross-sectional view of an automatic cell culture device equipped with a culture container transfer device according to an embodiment of the present invention; 3rd IQ is a machine room of the 9th Pigeon Cultivation Device shown in 11&C above; 11111, Figure 3 shows a section 1L of the loading/unloading device installed in the automatic culture device shown in 11111 above. Culture vessel transfer fct showing an example
flat msa. FIG. 1 is the main part of the Kuhai * device installed in the 9 automatic culture device shown in 1- above, No. 719! l is the 11th above! A perspective view of the main part of the II opening/closing device installed in the automatic culture device shown in IK - 1g5m is a side view of the peeling device installed in the Kyushu culture device shown in No. 111 above. Figure 11) is a partial cross-sectional view of the pipette dispensing device installed in the automatic culture device shown in Figure 1. Figure 11) is a cross-sectional view of the pipette supply device attached to the Kubu-eigin volume shown in Figure 119 above. FIG. 11 is an am#jt view showing the arrangement relationship between the pipette separating device and the pipette separation device shown in the above-mentioned No. 9@. FIG. 12 shows the rotation locus of the base pump in the dispensing device shown in FIG. 9 above, and the above! The pipette holder of the pipette supply device shown in Ito1m is a plan view of the parts shown in FIG. 11 above and the positional relationship with the cam lever of the pipette release device. A plan view of the petri dish feeding device installed in the apparatus, FIG. 14, is shown in FIG. 1 above, and a sectional view of the observation device installed in the automatic culture device, FIG. 11th enlarged perspective view of the objective lens driver S in the shown observation device;
wA is the flowchart showing the sequential process of cell culture in the Tomo's own culture * apparatus shown in No. 111 above.
...Automatic culture device 3...Culture room 4...Petri dish (culture container) 5...
... Carrying in/out device 6... Transfer silver amount 7... Waste liquid device 8... Feeding *i device 9. ......Peeling 1I placement 11...Dispensing device 18...Petri dish feeding device 18...
...Controlled wheel amount 14...Observation device 34...Motor 3b...Rotating disk (rotating member) 38m
......Spin shaft (-direct axis) 38...
・,
...Placement member 43...Plate spring member (elastic element R) 78.
......Lid opening/closing device 95...Pipeat supply device 96...
... Pipette intestinal evacuation device patent applicant Mr. Fujikawa, representative of Olympus Optical Industry Co., Ltd.
Seventh part horse 1 ward 47 62 fig. LI 2 ward Q et al. 614 ward

Claims (1)

[Claims] The motor (34) is arranged so as to be rotationally driven around the center of the straight shaft (llsm) and the nine rotating members (U, U).
), and a horizontal axis (319
) and a plurality of container mounting members (41, 42) which are swingably mounted around the container mounting member (41, 42) and interposed between the container mounting member (41, 42) and the rotating member (38). and the container mounting member (4
1, 42) to assume a horizontal position.
43) A culture vessel transfer device characterized by comprising: