JPH0724598B2 - Rotary magnetic shaking culture measurement method and device - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and apparatus for culturing various microorganisms in a culture test tube under magnetic shaking while measuring the growth rate of the microorganisms in parallel in a time-division manner and recording them.

[0002]

2. Description of the Related Art For example, Japanese Patent Publication No.
There are 3-43588 publication "Continuous automatic shaking culture apparatus and method" and JP-B-56-38195 "Rotation shaking culture measuring method and apparatus". Shaking means in the device of this kind from the past, is to shake the operation by directly applying a striking force to the culture tube itself, resulting in damage to the culture tube,
Moreover, the mechanism for shaking the culture tube separately was considerably complicated.

The invention of JP-B-53-43588 is
By arranging a photoelectric colorimeter at an appropriate place, the growth rate of the bacteria in all the culture tubes is measured at regular intervals. For that purpose, it is an essential condition that the culture tube is circulated along a guide rail forming a zigzag closed loop, and at the same time, a reciprocating shaking rod provided at the lower part of the guide rail is installed in the culture tube. The lower part is tapped, and the culture tube is shaken by the circular motion caused by the upper part as a fulcrum. Therefore, the device itself must be large-scale and complicated. Moreover, no improvement is seen in the drawback that the conventional culture tube is directly hit with force and damages the culture tube.

[0004]

[Problems to be Solved by the Invention] Further, Japanese Patent Publication No. 56-3
In the invention of Japanese Patent No. 8195, since an L-shaped culture tube is adopted instead of the usual straight rod-shaped culture test tube in order to avoid the above-mentioned drawbacks, it is very difficult to perform maintenance such as installation, removal and cleaning. Become. Further, when culturing anaerobic bacteria, it is necessary to avoid contact with oxygen in the air as much as possible, but in the case of the invention of Japanese Patent Publication No. 56-38195, during the culture of anaerobic bacteria. Even if it is possible to avoid contact with oxygen by leaving it in a stationary state, during the measurement and recording, the rotation of the inclined rotating disk causes the exchange of air between the horizontal part and the vertical part of the L-shaped culture tube. There is a drawback that the area is inevitably increased, which adversely affects the culture of anaerobic bacteria. Here, in view of the above-mentioned conventional drawbacks, the present invention is effective and suitable for stirring a culture containing anaerobic bacteria, and further, a rotary magnetic shaking culture measuring method and device which do not directly hit the culture tube. It is intended to be provided.

[0005]

The solution to the above-mentioned problems can be achieved by adopting the novel characteristic construction means listed below by the present invention. That is, the first feature of the method of the present invention is
While continuously orbiting the straight rod type culture test tube group standing along an inclined circular orbit surrounded by a continuous or intermittent magnetic field on the outer periphery, the culture test tube group with a magnetic stirrer inserted is revolved. While stirring the culture in the culture test tube by, while measuring the proliferation rate in the culture in each culture test tube approaching the appropriate position of the inclined circular orbit, the time-dependent measurement processing by the measurement value signal obtained at regular intervals. This is a rotary shaking culture measuring method.

The second feature of the method of the present invention is that the revolving circulation in the first feature is carried out by rotating the inclined turntable on which the culture test tube group is inserted and stopped in a continuous or intermittent indexing manner. This is a rotary magnetic shaking culture measurement method.

The third feature of the method of the present invention is that the concentration of each measured value in the culture in the first or second feature is obtained at a constant cycle as the tilting turntable turns. This is a rotary magnetic shaking culture measurement method using a synchronization signal.

A fourth characteristic of the method of the present invention is that the magnetic stirrer according to the first, second or third characteristic is changed in frequency of vibration applied thereto to control the strength of vibration of the stirrer. This is a rotary magnetic shaking culture method.

The first feature of the device of the present invention is that a tilting rotary disk, in which a group of straight rod-shaped culture test tubes are vertically arranged at desired intervals in the circumferential direction, is pivotally supported, and the tilting rotary disk is swung by a drive mechanism. While the shaking mechanism to be freely installed in a constant temperature bath, insert a magnetic stirrer in the culture test tube, on the other hand, arrange a continuous or intermittent magnetic body along the vicinity of the outer periphery of the inclined turntable,
While being configured to stir the inside of the culture test tube by rotation of the tilting turntable, the tilting turntable is equipped with a growth degree measuring device capable of time-divisionally measuring the growth rate of the culture in the culture test tube. It is a rotary shaking culture measuring device arranged in close proximity.

A second feature of the device of the present invention is a rotary magnetic shaking culture measuring device in which the intermittent magnetic substance groups in the first feature are arranged in an ascending or descending stepped ridge-valley shape.

A third feature of the device of the present invention is a rotary magnetic shaking culture measuring device in which the continuous magnetic bodies in the first feature are arranged in an arc shape, a wave shape, and a mountain valley shape.

A fourth feature of the device of the present invention is that the tilting rotary disc in the first, second or third feature is disc-shaped,
On the turntable, culture test tube groups stand on the same circle at equal intervals.
Is a rotary magnetic shaking culture measuring device reconnected hole groups corresponding conform to the seat tube group formed by penetrating through the inclined rotary disc with disposed at equal intervals seat tube to allow transmural stopped by Te .

The fifth feature of the device of the present invention is that, in the first, second, third, or fourth feature, the lower part of the culture test tube group is set up in the insertion hole group of the inclined rotary disk and stopped. While the light source and the photometer are held upside down in a radial direction with an appropriate position on the circular orbit, a sync signal pin is placed on the radius line connecting the center of the lower end of each insertion hole and the center of rotation of the lower surface of the inclined turntable. Rotation that is caused by the light source for synchronization and the photometer for synchronization signal relative to each other by sandwiching the circular orbit of the pin group for synchronization signal in the same radial direction as the relative radius direction of the light source and photometer on the other hand. It is a magnetic shaking culture measuring device.

[0014]

【Example】

(Example of Device) The device of the present invention will be described in detail with reference to the drawings. FIG. 1 is an explanatory view of the rotary magnetic shaking culture device of this embodiment, and FIG. 2 is FIG.
Fig. 3 is a plan view taken along line II-II with the culture test tube group inside removed.
FIG. 4 is another example of the arrangement of magnetic bodies along the vicinity of the outer periphery of the inclined turntable of the rotary magnetic shake culture device. In the figure, A is a rotary shaking culture measuring apparatus of the present embodiment, 1 is a tilting rotary disk, 2 is a drive mechanism for driving the tilting rotary disk 1, 3 is a shaking mechanism, 4 is a proliferation degree measuring unit, and 5 is a synchronization signal. Generation part, 6 is a fixed shaft ring, 7
Is a driver for revolving the inclined rotary disk 1 through the drive shaft 8 and the fixed shaft 6; 9 is a pin for synchronizing signal for obtaining a synchronizing signal; 10 is a light source for measuring the proliferation degree; 11 is a light intensity for measuring the proliferation degree. Reference numeral 12 is a light source for synchronizing signal, 13 is a photometer for synchronizing signal, 14 is a synchronizing signal generator, 15 is a recorder, 16 is a receiving tube for inserting and fixing the straight rod-shaped culture test tube a, and 17 is an inserting A hole, 18 is a proliferation degree measuring instrument, a is a culture test tube, b is a magnetic stirrer, c to c ″ are magnetic substances arranged along the vicinity of the outer circumference of the inclined rotary disk 1, s1 is a concentration measurement value signal, and s2 is Is a synchronization signal.

[0015] Specifically, rotary shaking culture measuring device A of this embodiment, upright every regular intervals culture tube a pivoted inclined rotating disk 1 capable array, the inclined turntable 1 drive mechanism The shaking mechanism 3 which can be swung by 2 is installed in a thermostatic chamber (not shown), and a proliferation degree measuring device 18 which measures the recording degree of the proliferation degree in the culture tube a which changes moment by moment in proximity to the inclined turntable 1 is provided. It is arranged.

As shown in FIG. 1, the inclined turntable 1 has a disk shape, and a shaft fixing ring 6 is fixed to the central portion of the lower surface thereof, while the culture test tubes a are arranged at equal intervals in the outer peripheral area of the upper surface. The receiving cylinders 16 are arranged on the same circle so as to be possible. In addition, the group of culture test tubes a standing upright and stopped on the tilting rotary disc 1 appropriately projects the culture test tube lower part a1 from the tilting rotary disc 1 to provide the light source 10 for proliferation degree measurement and the photometer 11 for measuring proliferation degree. Of the synchronizing signal pins 9 having the same circular shape projecting on each radial line connecting the center of the lower end of the insertion hole 17 of the tilting rotary disk 1 and the center of rotation of the lower surface of the tilting rotary disk 1. It will be arranged.

Incidentally, the number of the culture test tubes a group arranged and fixed on the inclined turntable 1 can be arbitrarily set on the condition that they are arranged on the same circle and at equal intervals. It is sufficient to provide a number of receiving cylinders 16, insertion holes 17, and groups of synchronization signal pins 9.

The drive mechanism 2 has only to be constructed so as to give the tilting turntable 1 a turning motion. For example, a motor is adopted as the driving body 7, and a belt is connected between the pulleys fixed to the motor shaft and the driven shaft 8 respectively.
The driven shaft 8 of the motor shaft may be directly connected. Accordingly, if the rotation speed of the tilting rotary disk 1 is set to a desired value, the driving body 7 can give the tilting rotary disk 1 a continuous rotary motion. If necessary, if a stepping motor is adopted as the driving body 7 and intermittent operation is performed by a step control signal, the tilting rotary disk 1 can be given intermittent indexing rotary motion.

Next, the proliferation degree measuring device 18 comprises a proliferation degree measuring section 4 comprising a light source 10 and a photometer 11 for obtaining a concentration measurement value signal s1, a light source 12 and a photometer 13 for obtaining a synchronizing signal s2, and A synchronizing signal generator 5 including a synchronizing signal generator 14, and a recorder 15 for time-divisionally recording the growth rate of the cultures in all the culture tubes a at regular intervals by both signals s1 and s2. Composed of.

The proliferation degree measuring unit 4 is fixed so that the lower part a1 of the culture test tube group a passes through the middle of the light source 10 and the photometer 11. Then, after the light rays emitted from the light source 10 pass through the lower part a1 of the culture tube a, they are arranged on the same optical axis so as to be detected by the photometer 11. Also,
The transmitted light detected by the photometer 11 is photoelectrically converted and supplied to the recorder 15 as a density measurement value signal s1.

The sync signal generator 5 comprises a sync signal light source 12, a photometer 13 and a sync signal generator 14, which are arranged opposite to each other on the same optical axis.
When the light beam from is passed through the synchronizing signal pin 9 and received by the photometer 13 in accordance with the turning operation of the rotary tilting plate 1, the electrical signal photoelectrically converted by the light receiving section is supplied to the synchronizing signal generator 14. , As a result, the sync signal s from the sync signal generator 14
2 is sent to the recorder 15 and used as a time division control signal.

The recorder 15 records the degree of growth of the culture in the culture tube a, which is a detection target. The recorder 15 measures the concentration measurement value signal s1 detected at regular intervals and the synchronization signal s2.
It is time-division controlled and is displayed / recorded over time.
The shape of the magnetic stirring bar b inserted into the culture tube a may be any shape that does not hinder the stirring of the culture in order to make the culture concentration uniform.

The magnetic body c arranged on the outer circumference of the tilting rotary disk 1 may be a permanent magnet or an electromagnet. When an electromagnet is used, it is possible to stir the culture by exciting the stirrer with vibration by changing the frequency of the current. Further, the shape thereof can be an embodiment such as an intermittent rectangle as shown in FIG. 3 or an arc-shaped continuous body as shown in FIG. Further, in FIG. 1, the position where the magnetic material c faces the culture test tube a is at a height position substantially equidistant from the opening of the culture test tube a. It is also possible to desire further agitation by arranging it in a vertical direction such as a mountain valley shape.

(Example of Method) An example of the method applied to this example of the apparatus will be described with reference to FIGS. First, a sample, a medium, a microorganism and a magnetic stirrer b are put into the culture test tube a by appropriate means, and then the catch cylinder 16 of the tilting rotary disk 1 and the insertion hole 17 are arranged in a stop arrangement. Then, when the driving body 7 is started, the tilting turntable 1 is caused to perform a turning motion along the tilted circular orbit to revolve the culture test tube group a. With this swirling movement, the stirring bar b in the culture test tube a momentarily changes its position and posture in the culture test tube a due to the magnetic force of the magnetic body c that forms a magnetic field arranged on the outer circumference of the inclined rotary disk 1. Since it is changed, the culture is agitated without directly shaking the culture test tube a, and the magnetic shake culture is carried out.

By the way, in this method example, on the inclined turntable 1.
Distribute 60 culture test tubes a group at equal intervals in the circumferential direction of
Line up and rotate the tilt turntable 1 at a rate of 1 turn per minute
If so, the recorder 15 includes a photometer 13 for each culture test.
Whether the concentration measurement value signal s1 of the test tube a is the synchronization signal generator 14
Then, the synchronization signal s2 is transmitted every one second. That
Then, the recorder 15 outputs the signals s1 and s sent one after another.
Based on 2, at a cycle of once a minute for each culture test tube a
The density measurement value signal s1 is sampled and appropriately processed.
To go. One rotation of the inclined turntable 1 is in descending order of high and low under the action of centrifugal force.
Inducing the seesaw state of the line, and
At the same time as imparting levitation and sinking forces to living things etc. at the same time,
In cooperation with agitation near the liquid surface due to fluctuations accompanying the rotation of the liquid surface
Quickly promote uniform densification of the whole.

Therefore, the concentration measurement value signal s1 obtained at regular intervals makes it possible to observe the growth rate of microorganisms in all the culture test tubes a in the magnetic shaking culture over time. In addition, in the culture of anaerobic bacteria, leave it stationary during the culture, and only at the time of measurement recording, the inclined rotary disk
By performing the intermittent indexing motion that causes the swirling motion, the culture is stirred and the contents are homogenized, so that accurate growth rate data of the culture over time can be obtained.

[0027]

As described above, according to the method and apparatus of the present invention, the shaking mechanism 3 is simplified and the production is facilitated as compared with the conventional one, and the culture test tube can be used without direct impact. Since it is possible to stir and the modified culture test tube is not used, it is very easy to attach and detach the culture test tube, so the work efficiency is improved, and moreover, the anaerobic bacteria It exhibits excellent usefulness in that it can also exhibit excellent effects in the culture and recording measurement.

[Brief description of drawings]

FIG. 1 is an explanatory view of a rotary magnetic shaking culture device showing an embodiment of the present invention.

FIG. 2 is a plan view taken along line II-II with the culture test tube group in FIG. 1 removed.

FIG. 3 is another example of the arrangement of magnetic materials along the vicinity of the outer periphery of the inclined rotary disk of the rotary magnetic shake culture device showing the embodiment of the present invention.

FIG. 4 is still another example of the arrangement of magnetic bodies along the vicinity of the outer periphery of the inclined rotary disk of the rotary magnetic shaking culture device of the present invention.

[Explanation of symbols]

 A ... Rotary shaking culture measuring device 1 ... Inclined rotary disk 2 ... Drive mechanism 3 ... Shaking mechanism 4 ... Proliferation degree measurement unit 5 ... Synchronization signal generation unit 6 ... Axis fixed ring 7 ... Driving body 9 ... Synchronization signal pin 10 ... Light source for proliferation degree measurement 11 ... Photometer for proliferation degree measurement 12 ... Light source for synchronization signal 13 ... Photometer for synchronization signal 14 ... Synchronization signal generator 15 ... Recorder 16 ... Receiving cylinder 17 ... Insert hole 18 ... Proliferation degree measurement Container a ... Culture test tube b ... Magnetic stirrer c-c "... Magnetic material s1 ... Concentration measurement value signal s2 ... Synchronous signal

Claims (9)

[Claims] 1. A culture test tube in which a magnetic stirrer is inserted while sequentially revolving a straight rod-shaped culture test tube group standing along an inclined circular orbit surrounded by a continuous or intermittent magnetic field. While stirring the culture in the culture test tube by the orbital movement of the group, the degree of proliferation in the culture in each culture test tube approaching the appropriate position of the inclined circular orbit, a measurement value signal obtained at regular intervals The method for measuring culture by rotary magnetic shaking, which is characterized by being subjected to time-dependent measurement processing by. 2. The rotating magnetic shaking culture according to claim 1, wherein the revolving circulation is carried out by rotating an inclined turntable in which a test tube group is inserted and stopped in a continuous or intermittent indexing manner. Measuring method 3. The concentration of the measured value signal and the synchronization signal, which are obtained at a constant cycle along with the turning of the inclined turntable, are used for the different degrees of growth in the culture, and the degree of proliferation is utilized. Rotary magnetic shaking culture measurement method. 4. The rotary magnetic shaking culture method according to claim 1, wherein the magnetic stir bar controls the vibration intensity of the magnetic stir bar by changing the frequency of the applied vibration. 5. A shaking mechanism in which a tilting rotary disk having vertical rod-shaped culture test tube groups arranged upright at desired intervals in the circumferential direction is rotatably supported, and each tilting rotary disk is rotatable by a drive mechanism in a constant temperature bath. On the other hand, while installing the magnetic stirrer in the culture test tube ,
On the other hand, a continuous or intermittent magnetic material is arranged along the vicinity of the outer periphery of the inclined rotary disc, and the rotation of the inclined rotary disc is configured to agitate the culture test tube while the inclined rotary disc is provided. A rotary magnetic shaking culture measuring apparatus, in which a growth degree measuring device capable of time-divisionally measuring the growth degree of the culture in the culture test tube is placed in close proximity. 6. The rotary magnetic shaking culture measuring apparatus according to claim 5, wherein the intermittent magnetic substance groups are arranged in a step-like ridge-valley shape that rises or falls. 7. The rotary magnetic shaking culture measuring apparatus according to claim 5, wherein the continuous magnetic bodies are arranged in an arc shape, a wave shape, and a mountain valley shape. 8. The tilting rotary disk has a disk shape, and the receiving cylinders are arranged at equal intervals on the rotary disk so that the culture test tube groups can stand up on the same circle at even intervals. The rotary magnetic shaking culture measuring device according to claim 5, 6 or 7, characterized in that a group of insertion holes corresponding to the group of receiving cylinders are provided through the inclined turntable. 9. A light source and a photometer are placed relative to each other in a radial direction with a proper position on a circular orbit of a lower portion of a culture test tube group which is erected and stopped by a group of insertion holes of an inclined turntable, while said A synchronizing signal pin is projected on a radial line connecting the center of the lower end of each insertion hole and the center of rotation of the lower surface of the inclined turntable, and on the other hand, the group of synchronizing signal pins in the same radial direction as the relative radius direction of the light source and the photometer. 9. The light source for synchronization and the photometer for synchronization signal are relatively placed with the circular orbit of 5 in between.
The rotating magnetic shaking culture measuring device described.