JPH0140649B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to simultaneous, controlled, non-intrusive mixing of a plurality of substances in individual containers, and in particular automatic mixing of the above-mentioned substances and the accompanying activation of an automatic analyzer. The present invention relates to a novel method and apparatus.

There are many types of mixing methods and devices known in the art that operate by immersing a mechanical part, such as a paddle or stirring piece, invasively into the substances to be mixed. There is. Such conventional mixing methods and devices have not been very satisfactory for use in current automated sample analyzers, such as those using particulate reagents. In particular, such invasive mixing devices (a) introduce cross-contamination between each sample by introducing into subsequent samples residues from previous samples that remain in the mixing device, and (b) The sample is diluted by introducing into the mixing device the remainder of the washing solution remaining following its washing, as described in U.S. Pat. No. 3,134,263 to Eduard B.M. de-Jong; or (c) interrupting a relatively sensitive chemical reaction by mechanical interference, or paragraphs (a), (b), and (c) above occur together, thus impairing the accuracy of the analytical product. It acts in a way that has a negative impact on the Furthermore, some automatic analyzers require continuous mixing of multiple sample-reagent combinations in individual open containers supported on indexing carousels, thus requiring multiple invasive mixing devices. Operational use is completely impractical.

Furthermore, although some non-intrusive mixing methods and mixing devices are known in the art for mixing substances by producing one or more movements of a container, to the best of the inventor's knowledge no automatic analytical device There is nothing particularly suitable for use. For example, U.S. patent no.
The paint mixing method and mixing device described in specification No. 3542344 is limited in its action to a closed container that must be manually placed on and removed from the mixing device. It is clearly unsuitable for use in the continuous and simultaneous mixing of separate open and unrelated sample containers. Similarly, the sample processing apparatus described in U.S. Pat. It only acts to vibrate the container intermittently, for example at the end of each indexing operation.
The mixing effect obtained in this case is not very satisfactory for use in automatic analyzers using granular reagents, as contemplated in one application of the present invention. Also, US Patent No.
In the liquid testing method described in specification No. 3528544,
A plurality of sealed ampoules are rotatably disposed and an indexable rotary table is used, which is intermittently rotated in different directions and at different speeds based on selected groups by a plurality of belt drive members. Also, the complexity of the multiple belt drive segments and the structure for holding and supporting the ampoules, as well as intermittent rotations, make such devices less than satisfactory for use in automated analysis systems such as those described above.

It is therefore an object of the present invention to provide a new method and apparatus for the continuous, controlled and non-intrusive mixing of particulate material into a liquid medium.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a method and apparatus adapted for use with automated analytical equipment requiring continuous and simultaneous mixing of one or more particulate materials or samples in a liquid medium in controlled mixing proportions. We are trying to provide the following.

It is a further object of the present invention to provide a method and apparatus for simultaneously supporting particle agglutination immunoassays in a plurality of containers supported on an indexable carousel.

It is an object of the present invention to provide a method and apparatus that uses only readily available parts that are relatively simple in form and operation and have proven reliability, minimizing costs and maximizing reliability. It's for a purpose.

The method and apparatus according to the invention allows for continuous mixing of substances contained in a generally cylindrical container having a plurality of open tops arranged in a generally circular array on an indexable circular carousel. Each of these containers is supported on a turntable for rotation about its axis.
This turntable forms part of an automatic sample analyzer which can serve for quantitative analysis of samples reacting in individual containers, for example by the particle agglomeration method described below. To effect the mixing, each container is moved around its axis in alternating opposite directions by driving a drive disk in frictional contact with each container in successive alternating directions, i.e., clockwise and counterclockwise. simultaneously rotate at a predetermined rotational speed for a predetermined time period. Reversal of the rotation of the vessel is continuous during indexing of the carousel, accelerating the completion of the reaction and thereby maximizing the precision of the analytical composition. Such mixing methods have particular applications, for example, in using particulate reagents to perform particle agglutination immunoassays. It has been found that such mixing very substantially increases the probability of contact of the particulate reagents and accelerates the completion of the reaction.

Embodiments of the mixing method and mixing apparatus according to the present invention will be described in detail below with reference to the accompanying drawings.

As shown in FIGS. 1 and 2, according to a preferred embodiment of the present invention, an indexable sample collection device 1
0 includes a central shaft 12 that is supported so as not to rotate from a fixed support base 14 and a fixed support plate 16.

The indexable circular turntable 18 can be rotated intermittently relative to the shaft 12 by an indexing drive 20.
The index drive device 20 includes a drive boss 2 supported for rotation with respect to the shaft 12 by a thrust bearing 24.
2. The drive sleeve 26 is rotatably mounted on the shaft 12. The lower and upper ends of the sleeve 26 are fixed to the drive boss 22 and the lower side of the rotary table 18. The helical gear 28 is provided around the drive boss 22 and meshes with a drive worm 30 driven by an intermittent electric motor 32. Based on this structure, the rotary table 18 is intermittently rotated or indexed around the support shaft 12 as shown by the arrow α in FIG. 1 by periodically energizing the intermittent electric motor 32.

The roughly circular arrangement of the reaction tube mounting holes 34 corresponds to the rotating table 1.
8 and adjacent thereto as shown in the figure. Within each bore 34 is a generally cylindrical lip bearing 3 made of a suitable low friction material such as nylon.
6 is placed.

The annular support shelf 38 is connected to the turntable 18 by circumferentially spaced support pieces 40 as shown.
It is supported by The support shelving 38 may be mounted to the support piece 40 in such a manner that it can be easily removed for normal disassembly of the sampling device 10.

Each sample container or reaction tube 42 is disposed within a bearing 36 for rotation about its axis either clockwise or counterclockwise as indicated by arrow β in FIG. The bottom of the reaction tube 42 is connected to a support shelf 3.
It fits within a molded recess 44 formed in the top surface 46 of the 8. Support shelf 38 is preferably constructed from a low friction material, such as nylon, to minimize friction and the associated risk of undesired heating of the reaction tube. Alternatively, cup-shaped bearings (not shown) may be inserted into the low-friction support recesses 44 as described for the reaction tubes 42 in each example.

Rotating device 50 operable in accordance with the present invention to produce precisely controlled two-way rotation of reaction tube 42 includes a driven pulley 52 and an interlocking drive boss 54 mounted for rotation as a unit about drive sleeve 26 as shown. It is equipped with Pulley 52 rides on the upper surface of drive boss 22 such that both of them are completely free to rotate relative to each other.

The annular drive disk 56 is secured to the boss 54 by a mounting ring 58 and spaced apart clamping pieces 60 as shown.
It is fixed at Drive disk 56 is made of a suitably rigid material with high surface friction properties, such as silicone rubber. As illustrated in FIGS. 1 and 2, the periphery of the drive disk 56 is securely in contact with the side wall of each reaction tube 42, and as a result of the clockwise rotation of the drive disk 56 in FIG. The reaction tubes 42 are rotated counterclockwise around each axis at the same rotational speed. Further, as the driving disk 56 rotates counterclockwise in FIG. 1, all the reaction tubes 42 rotate clockwise. Because the diameter of the drive disk 56 is extremely large, the rotational speed of each reaction tube 42 in all examples does not require potentially difficult high-speed rotation of the driven pulley 52, drive boss 54, and drive disk 56, as will be described in detail below. The rotational speed of the drive disk 56 is substantially higher so that the substances in each reaction tube 42 are sufficiently mixed.

With the exception of thrust bearing 24, which is not shown, it will be apparent to those skilled in the art that suitable bearings may be provided as desired for all relatively rotatable parts of sampling device 10.

A reversible variable speed electric motor 62 operates to rotate a drive pulley 64. The drive belt 66 connects the drive pulley 64 to the driven pulley 52 and connects the drive pulley 64 to the driven pulley 52.
rotates as the drive motor 62 is energized. Therefore, due to the energization of the drive motor 62, the driven pulley 52,
When the drive boss 54 and the drive disk 56 are driven, the reaction tubes 42 are simultaneously rotated around their respective axes as described above. The substantial difference in the diameters of the pulleys 64, 66 allows the drive motor 62 to operate at higher torque speeds which are more effective without overspeeding the drive disk 56 or reaction tube 42. The drive motor torque amplification factor resulting from the difference in the diameters of the respective pulleys allows very rapid deceleration and acceleration of the drive disk 56 and thus the reaction tube 42 between opposite rotational speeds, as will be explained in more detail below.

Drive motor control device 6 diagrammatically shown in FIG.
8 is operable to adjustably and precisely control the rotation speed, rotation direction, and rotation time in each direction of the drive motor 62. Therefore, the rotation speed, rotation direction, and rotation time in each direction of each reaction tube 42 can be precisely controlled over a wide range by appropriate adjustment of the control device 68.

For example, the sample collection device 10 can be effectively applied to an automated device that performs particle agglutination counting immunoassays. Such automated equipment quantitatively analyzes multiple serum samples for their free thyroxine levels. This free thyroxine level indicates the thyroid hormone level of each serum sample. Such analysis is referred to as a one-particle methodology by placing a precisely predetermined, relatively small volume of appropriately diluted serum sample into each reaction tube 42. Each such sample volume is provided with a suitable antibody-coated agglutination medium, such as a latex sphere approximately 1 micron in diameter coated with a rabbit anti-thyroxine antibody for the serum sample component of interest, and a predetermined volume of a suitable similar specific antigen or tracer. For example, a predetermined volume of particulate reagent consisting of a non-particulate soluble polymeric material such as dextran which acts as a specific binding agent to form bridges between the specific antibody-coated latex spheres is thoroughly mixed. Optically detectable aggregation of this reagent therefore occurs. This aggregation is inversely proportional to the serum free thyroxine level in the serum sample in question. That is, serum free thyroxine competes with antigen for binding to antibody-coated latex spheres. In this case, the higher the concentration of thyroxine, the lower the total level of latex sphere aggregation.

To maximize the precision of the analytical composition, the particulate reagents or latex spheres must be bound to each other by specific antibody-antigen bridges to a maximum degree comparable to the serum sample thyroxine level. The latex spheres, which have a diameter of approximately 1 μm and therefore do not inherently move rapidly by diffusion, must therefore be driven into intimate contact in the reaction mixture. Furthermore, since the latex sphere concentration in the reaction mixture is relatively low, the distance between each latex sphere is relatively large and the rate of natural contact between these latex spheres per unit time is very small. For example, if a reaction mixture is left unmixed, the time for the aggregation reaction to reach typical steady state or equilibrium conditions can be as long as 4 hours. Further, if this same solution is constantly mixed by stirring so that equilibrium occurs between each latex sphere, such time can be reduced to less than 30 minutes. However, this mixing must be gentle enough so that the latex spheres, once agglomerated, do not separate again due to the mixing action, yet strong enough to reach a steady state or equilibrium within the shortest practical time. There must be. Thus, it is clear that substantial care must be taken in determining the nature and proportions of the mixture.

As shown in FIGS. 1 and 2, the serum sample inhalation needle, antibody-coated latex sphere (granular reagent), antigen (tracer) and buffer dispensing needle, and reacted sample solution suction needle each have a needle 70. ,72,7
4, 76, 78 and are operatively disposed relative to sampling device 10. Each dispensing needle 70,7
2, 74, and 76 operate non-invasively to sequentially dispense a predetermined volume of the indicated substance into each reaction tube 42.
The suction needle 78 then serves to remove or aspirate a predetermined volume of reaction mixture from each reaction tube 42 for subsequent analysis.

The automated sample solution fractionator 82 includes an optical housing 84 containing an optical detection device 86, shown in dashed lines.
It is equipped with The detection device 86 includes a flow cell 88 . A reacted sample solution pump 90 is also provided, and a conduit 94 runs from the reacted sample solution suction needle 78 through the pump 90 to the flow cell 88 as shown in the figure.
and a predetermined volume of the reacted solution to each reaction tube 42.
through a suction needle 78 and pump 90 to a flow cell 88 as shown and then to a flow cell 88 as shown.
Along with this, automatic chemical analysis is performed by an optical detection device 86. The recording device 92 for recording the sample solution analysis composition may be, for example, a strip diagram recorder as shown. The recorder is operative to sequentially record the performance of each sample solution analysis, generally as shown, for example by curve 96.

In operation, the two-way rotation device 50 is energized to concomitantly rotate all reaction tubes 42 at predetermined speeds clockwise and counterclockwise during each successive time period controlled as described above. Also, when the indexing drive device 20 is energized, the rotary table 18 is periodically advanced clockwise as shown. Each rotating reaction tube 42 is indexed relative to the sample inhalation needle 70, the antibody-coated latex sphere dispensing needle 72, and the antigen dispensing needle 74 and a carefully predefined volume of these materials is added to the reaction tube 42.
sequentially and non-intrusively. The rotation of each reaction tube 44 causes sufficient but non-destructive mixing of such materials to promote aggregation reactions. Especially when the reaction tube rotation in one direction reaches its predetermined speed,
A vortex is generated within the reaction tube 42, and a mixing action is accordingly performed. However, when the direction of rotation of the reaction tube is abruptly reversed, this vortex is eliminated by the frictional force exerted on such mixture by the inner wall of the reaction tube 42. The reaction tube 42 then accelerates rapidly in the opposite direction, and this rotation of the reaction mixture in the other direction tends to create vortices and eventually causes the vortices to form again.
That is, each reversal of the direction of rotation of the reaction tube 42 creates a tearing force within the reaction mixture. This tearing force is sufficient to create a sufficient mixing action to promote the flocculation reaction, but is insufficient to separate the already flocculated latex spheres. The rotational speed of the reaction tube 42 is carefully predetermined in each case such that it is below the rotational speed at which each sample solution spills out of the reaction tube 42 due to the mixing action described above.

Subsequently, the carousel 18 is placed against the buffer dispensing needle 72 (to ensure that the addition of buffer to the reaction tube 42 produces a standard curve by the optical detection device 86), which is then replaced by the aspirating needle. 78 and advance each reaction tube 42 to continuously mix the sample solution as described above. In this case, a predetermined volume of the well-mixed reaction mixture is aspirated for delivery to the optical detection device 86 . Detection device 86 serves to measure the extent of latex globule agglutination within the serum sample and thus the serum free thyroxine level. Recording of the results of this measurement for each serum sample in question is performed by an analytical product recorder 92.

The reaction mixture placed in each reaction tube 42 is indexed on the turntable 18 from the time the various components are introduced into the reaction tube 42 until the internal volume of this reaction tube is removed for optical analysis as described above. It is especially important to mix the mixture continuously. The aggregation reaction is therefore continuously promoted without interruption, accelerating the reaction and reaching a steady state or equilibrium within a minimum time period.

A non-limiting diagram 80 of the speed and direction of rotation of reaction tube 42 versus time is shown in FIG. It is clear that by adjusting the drive motor controller 68, the direction of rotation of the reaction tube 42 can be reversed every about 1.5 seconds to achieve a peak rotational speed of about 1200 rpm in each direction. Furthermore, the typical residence time, ie the time between the introduction of antibody-coated latex spheres (granular reagent) and antigen (tracer) to initiate the agglutination reaction and the subsequent withdrawal of a volume of the arriving mixture for optical analysis, is It takes about 15min.

Although the case where the present invention is applied to the mixing of biological samples and the associated operation of an automated analyzer has been described in detail, it is clear to those skilled in the art that the present invention is not limited to such applications.

Although the present invention has been described in detail with reference to its embodiments, it is obvious that the present invention can be modified in various ways without departing from its spirit.

[Brief explanation of drawings]

FIG. 1 is a partially cutaway plan view showing an embodiment of the mixing device of the present invention, diagrammatically illustrated sample and reagent suction and suction parts, and a sample analyzer; FIG. FIG. 2 is a sectional view taken along line 2-2 of FIG. FIG. 3 is a diagram of reaction tube rotation speed versus time. 12... Central axis, 18... Rotating table, 20... Indexing drive device, 42... Reaction tube (container), 50...
... Rotating device, 56 ... Annular drive disk, 62 ... Drive electric motor, 68 ... Drive motor control device.

Claims (1)

[Scope of Claims] 1. A method for the controlled non-intrusive mixing of substances contained in a plurality of containers supported for rotation about the axis of each container on a rotatable support member, comprising:
A container driving device of each container simultaneously rotates each container in one direction around the container axis at a first predetermined speed for a first predetermined period of time, and alternating with simultaneous driven rotation in opposite directions at a second predetermined speed for a second predetermined period of time;
A method of mixing consisting in causing the mixing of the substances in each of said containers to take place in a constantly changing direction. 2. Using an indexable carousel as a rotatable support piece, the driven rotation of each container continues the driven rotation of each of said containers during both the indexing and dwell time periods of said carousel. 2. A method as claimed in claim 1, in which the mixing of the substances in the container continues in a constantly changing direction during operation of the turntable. 3. The mixing method according to claim 1, wherein the first and second rotational speeds are substantially equal to each other. 4. The mixed method according to claim 1, wherein the first and second predetermined time periods are substantially equal to each other. 5 The top of each container is opened, and the first and second rotational speeds are in each case less than or equal to the container rotational speed at which the substance spills from the open-topped container by mixing the substance. Mixed method as described in. 6. A rotating table is generally circular, and a plurality of containers are arranged on the rotating table in a substantially circular arrangement, and the plurality of containers are arranged in a substantially circular arrangement, and the driven rotation of each container moves the plurality of containers in the first and second directions. 3. A method as claimed in claim 2, in which the containers are associated in a concomitant driving manner so as to rotate. 7. The mixing method of claim 2, wherein alternation of the rotation direction of each container occurs approximately every 1.5 seconds, and the predetermined container rotation speed in each of these directions is approximately 1200 rpm. 8. The method of claim 2, wherein a predetermined amount of the substance is sequentially and non-intrusively dispensed into the containers when each container is indexed in turn to the appropriate position by means of a turntable. 9. The method of claim 2, wherein each container is sequentially indexed to an appropriate position by a turntable at the end of mixing of the materials in each case, and predetermined amounts of the material are sequentially removed from these containers. 10. A mixing method according to claim 9, in which the amounts of the mixed substances taken out are sequentially fed into an analytical device for analysis of these mixed substances. 11. A mixing device for performing controlled, non-intrusive mixing of substances contained in a plurality of separate containers, comprising a rotatable support piece that supports each of the plurality of containers in relative rotation; The containers are simultaneously rotated about each container axis in a first direction at a first predetermined speed for a first predetermined time period, alternating with this rotation, the same containers are rotated about each container axis in a first direction.
containers configured to simultaneously rotate in a second direction opposite to the above direction at a second predetermined speed for a second predetermined period of time so that the substances in each container are constantly changed in direction; and a rotating device. 12 The rotatable support piece is constituted by a freely indexable turntable, and a container rotating device acts to rotate each container during both the indexing and retention periods of the turntable, so that the inside of each container is rotated. 12. The mixing device according to claim 11, wherein the mixing of the substances is carried out continuously by constantly changing directions during each turntable operation. 13. The mixing device according to claim 11, wherein each container has an open top. 14. The mixing device according to claim 11, wherein the first and second rotational speeds are substantially equal to each other. 15. The mixing device according to claim 11, wherein the first and second time periods are substantially equal to each other. 16. Claim 1, wherein a plurality of containers are arranged in a generally circular arrangement on a rotating table, and the container rotating device is provided with a piece drivingly associated with each container.
2. The mixing device according to 2. 17. comprising a dispensing needle operatively cooperating with the turntable;
When each container is sequentially indexed by the dispensing needle to each position relative to the dispensing needle by the rotary table, a predetermined amount of substance is sequentially and non-intrusively dispensed into each of the containers. A mixing device according to claim 12, which is configured as follows. 18 The turntable is provided with an operatively cooperating suction needle by means of which each container is indexed sequentially to each position relative to said suction needle by said rotation at the end of the mixing of the substances in each instance and from each said container. 13. A mixing device as claimed in claim 12, adapted to take out predetermined quantities of the substance sequentially. 19 In each example, the first and second predetermined rotational speeds are
14. The mixing device according to claim 13, wherein the mixing speed of the container is set to be lower than the speed at which the substance spills from the open top container. 20. The mixing device according to claim 12, wherein the rotary table is flat, and each container is provided with axes of rotation that are parallel to each other and substantially perpendicular to the plane of the rotary table. 21. The mixing device according to claim 12, wherein the rotation speed of each container can be adjusted by driving the container rotating device. 22 Each container is generally cylindrical and has approximately equal outer diameter, and the container rotating device is provided with a generally circular drive disk that is in surface contact with the outer wall of each container and is arranged generally concentrically with the container array. 17. The mixing device of claim 16, wherein driven rotation of the drive disk in one direction causes a concomitant driven rotation of each container in the opposite direction about the container axis. 23. A mixed substance analyzer operatively connected to a suction needle for sequentially supplying predetermined amounts of substances from said suction needle to said analyzer for sequential analysis by said analyzer. A mixing device according to range 18. 24 A support shaft, a generally circular rotary table rotatably supported on the support shaft, and a rotary table indexing device rotatably supported on the support shaft and connected to exert a driving action on the rotary table. a rotating device that intermittently rotates the rotary table indexing device to index the rotary table; a plurality of generally cylindrical containers of equal dimensions each supported on the turntable in an extending manner for rotation about a container axis;
The peripheral portion of the drive disk is supported by the support shaft below the rotary table so as to rotate approximately concentrically therewith, and the peripheral portion of the drive disk extends below the rotary table into surface contact with the outer wall of each container with a driving action. a generally circular container driving disk having a diameter of 100 mm, and rotating the driving disk alternately in opposite directions about the support shaft to move each container simultaneously in alternating opposite directions about the container axis. a container-driving disk rotating device configured to rotate and operate during both indexing and retention periods of the rotating table to continuously change direction and continuously mix the substances in the container; , a mixing device for the controlled, non-intrusive mixing of substances.