EP3003545A1

EP3003545A1 - Method and apparatus for reagent mixing

Info

Publication number: EP3003545A1
Application number: EP14803986.0A
Authority: EP
Inventors: Steen Guldager Petersen; Armen VAHEBZADEH; Bent Pedersen
Original assignee: Dako Denmark ApS
Current assignee: Dako Denmark ApS
Priority date: 2013-05-28
Filing date: 2014-05-27
Publication date: 2016-04-13
Also published as: WO2014190998A1; EP3003545A4

Abstract

A mixing device is disclosed that includes a base, a magnet post configured for insertion into a hollow portion of a reagent container, a stationary magnet mounted on the magnet post, and a mixing ball including a ferromagnetic material provided in the reagent container, wherein the reagent container may be moved back and forth on the magnet post. Another mixing device is disclosed that includes a housing, a drive mechanism, a drive magnet mechanically coupled to the drive mechanism, a coupling magnet magnetically coupled to the drive magnet, a mixing ball including a ferromagnetic material provided in a reagent container, and a controller, wherein the controller is configured to control the drive mechanism to cause the drive magnet to move, and wherein the motion of the drive magnet induces a first corresponding motion in the coupling magnet, and wherein the first corresponding motion of the coupling magnet induces a second corresponding motion in the mixing ball,

Description

METHOD AMD APPARATUS FOR REAGENT MIXING

DESCRIPTION

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[001 ] Aspects of the present disclosure relate to apparatuses and methods for mixing reagent and buffer solutions prior to their use in laboratory assay procedures.

SUMMARY

[002] In one embodiment, an apparatus for manually mixing reagent and buffer solutions is provided, and may include a mixing ball and a stationary magnet.

[003] In another embodiment, a manual mixing apparatus may be partially automated, for example by mechanically or otherwise moving the reagent vial up and down, or mechanically or otherwise moving the magnet post up and down.

[004] In still another embodiment, an automated mixing apparatus may include a drive magnet and coupling magnet to facilitate motion of a mixing ball, A thermal element may be provided, and a controller may be included to control each component.

[005] In still another embodiment, an automatic mixing apparatus may be integrated with an automated assay instrument.

BRIEF, .DESCRIPTION OF THE DRAWINGS

[006] Fig. l a illustrates a mixing apparatus according to an exemplary embodiment of the disclosure.

[007] Fig. 1 b illustrates the use of a mixing apparatus according to an exemplary embodiment of the disclosure,

[008] Figs. 2a and 2b illustrate an exterior of automated mixing apparatus 200. [009] Figs. 3a and 3b illustrate exemplary embodiments of drive mechanism

240,

[010] Fig. 4 illustrates an automated mixing apparatus utilizing a computer for aspects of control.

[01 1] Fig. 5a illustrates an integrated automated mixing apparatus provided with an automated assay instrument.

[012] Fig. 5b illustrates a close up of a portion of an integrated automated mixing apparatus provided with an automated assay instrument.

DESCRIPTION OF THE EMBODIMENTS

[013] in some laboratory sample processing operations, for example, staining or in-situ hybridization, reagents may be used which may require mixing prior to dispensing the reagents for processing the sample. One property that may lead to a requirement for mixing is the tendency for some reagents to separate into component phases. In some cases, phase separation may occur gradually over time. Refrigeration or freezing of reagents may accelerate or increase the phase separation.

[014] Some reagents such as molecular probes for in situ hybridization are precisely diluted in a specified quantity of buffer. In situ hybridization (ISH) probes may be dispensed in volumes as small as 10 micro liters per specimen. Phase separation prior to or during dispensing of reagent, may change the relative ratio of probe to diluent, e.g. buffer, which may lead to inconsistencies in the quality of assay results.

[015] Moreover, reagent containers for ISH probes may be small volume containers that make it difficult to apply mixing techniques suitable for containers of larger volumes. For example, the narrow cylindrical diameter of an ISH reagent container may prevent application of mechanical stirring with a rod. Further, shaking or vibrational mixing may not be effective for very small volumes of relative viscous ISH reagent.

[016] in biological sample processing, even a minute amount of residual ISH reagent, e.g. DMA probe, left on a mechanical mixing mechanism can negatively impact assay results by contaminating a subsequent sample, in a diagnostic setting, a false positive result may have dramatic consequences and a false negative result may be even worse. Therefore, a mixing technique that eliminates the possibility of transferring even miniscule amounts of reagent from sample to sample is extremely important.

[017] Moreover, if a mixing mechanism reacts chemically or immunochemical^ in any way with either the sample or the assay, adverse results may occur. Hence, the mixing mechanism may ideally be nonreaciive with reagents and samples.

[018] The present disclosure describes the various methods and apparatuses utilizing a mixing bail susceptible to magnetic attraction to provide mixing through an up and down motion.

[019] Fig. 1 a illustrates a mixing apparatus according to an exemplary embodiment of the disclosure. Mixing apparatus 100 may include base 1 10, magnet post 120, and stationary magnet 130.

[020] Mixing apparatus 100 may be configured for use with a reagent container 140. Reagent container 140 includes housing 150, vial portion 160, and cap 170.

Housing 1 50 of reagent container 140 may be square or rectangular in cross section, or may have another cross-section shape suitable for use with automated sample assay equipment. Housing 150 may be hollow, with a closed portion at the top and an opening at the bottom. Vial portion 180 may be provided to contain reagent mixture 180, and may be formed integrally with housing 150 or separable from housing 150. Cap 170 may be provided to selectively seal and unseal vial portion 160. Via! portion 160 may be provided with mixing ball 190 to facilitate the mixing of reagent mixture 180 (not shown). Mixing hall 190 (not shown) may be magnetic Itself, or may include a material, such as a ferromagnetic material, attracted to magnets. Mixing ball 190 may also include a non-reactive plating, such as gold plating. Mixing ball 190 may be packaged in reagent mixture 180 inside of vial portion 160, or may be provided separately, and inserted into vial portion 180 when cap 170 has been removed. Mixing ball 190 may be a sphere. However, mixing ball 190 is not limited to this shape, and may also be oblong shaped, square shaped, pyramid shaped, or any other shape that may assist in mixing the reagent mixture. Mixing ball 190 may be smaller in diameter than a maximum diameter of vial portion 160, and may have a diameter (or, in the case of a non-circular shape, at least one dimension) of 20% to 99% of a diameter of vial portion 160.

[021] Fig. 1 b illustrates the use of a mixing apparatus according to an exemplary embodiment of the disclosure. A user may fit magnet post 120 of mixing apparatus 100 into the hollow bottom opening of housing 150. Such positioning may create a magnetic coupling between mixing ball 190 and stationary magnet 130. When reagent container 140 is moved up and down, as illustrated in Fig. 1 b, the movement of mixing ball 190 may be constrained by coupling the coupling with stationary magnet 130. Thus, when reagent container 140 is moved up and down with respect to stat onary magnet 130, mixing ball 190 moves up and down with respect to reagent mixture 180 contained within via! portion 160 of reagent container 140. The up and down movement of mixing bali 190 may cause the mixing of the separated components of reagent mixture 180. Although a particular arrangement of a reagent container 140 and mixing apparatus are discussed above, many alterations of may be realized without departing from the scope of this invention, For example, in an alternate embodiment, mixing apparatus may not include base 110, and may include magnet post 120 and stationary magnet 130 configured to be held in a users hand while the mixing occurs. In another aiternate embodiment, reagent container 140 may have an opening at the top of the reagent container, permitting a user to insert magnet post 120 and stationary magnet from above the reagent container 140,

[022] An exemplary method for using mixing apparatus 100 may proceed as follows. A user may retrieve a reagent container 140 from storage, for example, a refrigerator, freezer, or other type of supply closet. Due to storage conditions, reagent mixture 180 contained in reagent container 140 may require mixing before use. if the reagent container 140 has come from cold storage, it may be left in a room temperature environment to thaw and/or to warm to room temperature prior to mixing, if the reagent container 140 is not pre-packaged with a mixing bail 190, the user may insert one after removing the cap 170. Next, a user may utilize mixing apparatus 100, manually, to mix the reagent mixture 180. The user may cycle the reagent container 140 up and down on the magnet post 120 a predetermined number of times, for example 25, 50 or 100, and/or may cycle the reagent container enough times to visually confirm adequate mixing of reagent mixture 180. A user may cycle the reagent container up and down at a rate appropriate for the viscosity of the reagent mixture, for example, two cycles per second, one cycle per second, and/or one cycle per half second. Other rates may be used, depending on the composition of reagent mixture 180. After mixing, the user may place reagent container 140 Into an automated sample assay instrument for use.

[023] In another exemplary embodiment consistent with the present disclosure, mixing apparatus 100 may be supplied with a motor (not shown) in order to partially automate the above-described process. The motor may be mechanically coupled to magnet post 120 or to reagent container 140 in order to drive the required relative movement between the two. Such automation may permit a user to perform other tasks while the motor drives the mixing process.

[024] In another exemplary embodiment consistent with the present disclosure, a fully automated mixing apparatus 200 may be provided. Mixing apparatus 200 is illustrated in Figs. 2a, 2b, 3a, and 3b. Figs. 2a and 2b illustrate an exterior of automated mixing apparatus 200. Automated mixing apparatus 200 may include a housing 210, magnet post 220, coupling magnet 230, drive mechanism 240, drive magnet 250.

thermal element 260 (not shown), thermal sensor 265 (not shown), and controller 270 (not shown). After removal from storage, a user may place a reagent container 140 over magnet post 220, as illustrated in Fig. 2b, Automated mixing apparatus 200 may include sensors, such as pressure sensors or switches, to detect the present of reagent container 140 on magnet post 220. Thermal element 280 may be controlled by controller 270 to thaw and/or warm reagent mixture 80 to room temperature or to another specified temperature. Thermal sensor 265. which may be any type of commonly used thermal detection element, such as a thermistor, thermocouple, etc., may provide input to controller 270 for the thawing/warming process. [025] After thawing, controller 270 may control drive mechanism 240 to cause the mixing of reagent mixture 180. Figs. 3a and 3b illustrate exemplary embodiments of drive mechanism 240. As illustrated in Fig. 3a and 3b, drive mechanism 240 may be enclosed within housing 210. Such enclosure may protect the parts of drive mechanism from dust, dirt, and other contaminants, and may protect a user from the motor driven parts. When activated, the linkage of drive mechanism 240 may cause drive magnet 250 to travel up and down. While one particular structure of drive mechanism 240 is illustrated, a person of skill in the art may recognize and utilize other suitable

mechanism for propelling drive magnet 250 up and down without departing from the scope of the invention. When driven up and down, drive magnet 250, in turn, may cause coupling magnet 240, encased in magnet post 220 to also travel up and down. Controller 270 may be integrated within housing 210 of automated mixing apparatus 200, and automated mixing apparatus 200 may be provided with a display and control unit for user programming. Controller 270 may be configured to deactivate thermal element 260 and/or drive mechanism 240 if reagent container 140 is removed from magnet post 220.

[028] In some embodiments, automated mixing apparatus 200 may be powered by USB connection 310 to a computer 320, which may serve as and/or take the place of controller 270. Fig. 4 illustrates an exemplary embodiment including a computer 320, Controller 270 may provide visual reports on the mixing progress, such reports may be provided to a display unit Integrated with automated mixing apparatus 200, and/or to a computer monitor if a computer 320 is used for control. Controller 270 may provide alarms and/or alerts, either through a computer 320 or through dedicated lights, buzzers, etc., to notify a user that mixing has been interrupted. Controlier 270 may provide alarms and/or alerts, either through a computer 320 or through dedicated lights, buzzers, etc, to notify a user that mixing has been completed, in some embodiments, a scanner 330, such as a bar-code reader, may be provided. Scanner 330 may be utilized by a user to scan an identifier on reagent container 140 prior to placing the container. Scanning reagent container 140 may provide information to computer 320 and/or controller 270 regarding the contents of reagent container 140, Based on the contents of reagent container 140, computer 320 and/or controller 270 may select a suitable thawing and/or mixing protocol. Alternatively, scanner 330 may be integral with the housing 210 of automated mixing apparatus 200. and may be configured to scan reagent container 140 when it is placed over mixing post 220.

Furthermore, as illustrated in Fig. 4, computer 320 may be in communication with an automated assay instrument 370, and thus communicate information about the reagent container 140 being mixed as well as progress information about the mixing.

[027] In another exemplary embodiment consistent with the present disclosure, an automated assay instrument 400 may be provided with an integrated automated mixing apparatus 410. Fig. 5 illustrates an Integrated automated mixing apparatus 410 provided with an automated assay instrument 400. Integrated automated mixing apparatus 410 may include ail of the same features as described with respect, to automated mixing apparatus 200. An exemplary method of using integrated automated mixing apparatus 410 may proceed as follows. Automated assay instrument 400 may alert a user to the need to load a specific reagent container 140. A user may retrieve reagent container 140 from storage and place it in an appropriate position on integrated automated mixing apparatus 410. Alternatively, automated assay instrument 400 may have a loading station for insertion of reagent container 140 and robotic means to transport reagent container 140 to integrated automated mixing apparatus 410. Once placed, reagent container 140 may then be scanned to confirm thai is the correct reagent. Once confirmed, a controller of automated assay instrument 400 may control integrated automated mixing apparatus 410 to begin thawing, warming, and/or mixing of reagent mixture 180 stored in the placed reagent container 140. When the thawing, warming, and/or mixing of reagent mixture 180 is completed, automated assay instrument 400 may alert a user of completion and request that reagent container 140 be moved to a reagent utilization area of automated assay instrument 400.

Alternatively, robotic means may be utilized within automated assay instrument 400 to transport reagent container 140 to a reagent utilization area.

[028] Other embodiments of the present disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the present disclosure.

[029] While this disclosure provides examples of the mixing devices utilized for certain purposes, usage of the disclosed devices is not limited to the disclosed examples. The disclosure of uses of embodiments of mixing devices is to be

considered exemplary only. Furthermore, alternatives to the specific structures present herein may be utilized without departing from the scope of the invention. Alternative embodiments will become apparent to those skilled in the art to which the present invention pertains without departing from its spirit and scope. The following numbered paragraphs provide additional embodiments of the present invention.

Claims

WHAT IS CLAIMED IS:

1. A mixing device comprising:

a base;

a magnet post configured for insertion into a hollow portion of a reagent container;

a stationary magnet mounted on the magnet post; and

a mixing bail including a ferromagnetic material provided in the reagent container,

wherein the reagent container may be moved back and forth on the magnet post.

2. A mixing device comprising;

a housing;

a drive mechanism;

a drive magnet mechanically coupled to the drive mechanism a coupling magnet magnetically coupled to the drive magnet; a mixing ball including a ferromagnetic material provided in a reagent container; and

a controller,

wherein the controller is configured to control the drive mechanism to cause the drive magnet to move, and wherein the motion of the drive magnet induces a first corresponding motion in the coupling magnet, and wherein the first corresponding motion of the coupling magnet induces a second corresponding motion in the mixing ball.

3. The mixing device of claim 2, further comprising a thermal element.