Optical measurement device for reaction vessel and method therefor
11099132 · 2021-08-24
Assignee
Inventors
Cpc classification
G01N21/6452
PHYSICS
G01N21/6428
PHYSICS
G01N2201/0846
PHYSICS
G01N35/028
PHYSICS
G01N35/02
PHYSICS
International classification
G01N35/02
PHYSICS
Abstract
The invention relates to an optical measurement device for a reaction vessel, and a method therefor. An object is to measure the optical state within a reaction vessel in an efficient, rapid, and highly reliable manner, without an expansion of the device scale. The configuration includes: a vessel group in which two or more reaction vessels are arranged; a light guide stage having two or more linking portions to which front ends of light guide portions, which have a flexibility, that optically connect with the interior of the linked reaction vessels, are provided; a connecting end arranging body that has an arranging surface that arranges and supports along a predetermined path two or more connecting ends, to which back ends of the light guide portions, in which the front ends thereof are provided to the linking portions, are provided, the connecting ends are provided corresponding to the respective linking portions; a measurement device provided approaching or making contact with the arranging surface that has measuring ends that are successively optically connectable with the respective connecting ends along the predetermined path, and in which light from within the reaction vessels is receivable by means of optical connections between the connecting ends and the measuring ends; and a light guide switching mechanism that relatively moves the respective connecting ends arranged on the connecting end arranging body and the respective measuring ends such that they are successively optically connected.
Claims
1. An optical measuring device, comprising: a first optical fiber including opposing first and second ends; a second optical fiber including opposing third and fourth ends; a first reaction vessel comprising an opening to a first interior that is optically connectable with the first end of the first optical fiber; a second reaction vessel comprising an opening to a second interior that is optically connectable with the third end of the second optical fiber; a light guide stage comprising a light guide plate coupled to the first end of the first optical fiber and the third end of the second optical fiber; a nozzle head coupled to the light guide stage, the nozzle head comprising: a suction-discharge mechanism; a first nozzle connected to the suction-discharge mechanism; a second nozzle connected to the suction-discharge mechanism; a first dispensing tip detachably mounted to the first nozzle; and a second dispending tip detachably mounted to the second nozzle; wherein suction and discharge of gases by the suction-discharge mechanism causes suction and discharge of liquids by the first and second dispensing tips; and a controller operably connected to the light guide stage and the nozzle head; wherein the light guide stage and the nozzle head are driven by the controller in a vertical direction relative to the openings of the first and second interiors by a stage Z axis transfer mechanism to simultaneously optically connect: the first end of the first optical fiber with the first interior of the first reaction vessel, and the third end of the second optical fiber with the second interior of the second reaction vessel; a first measurement device for receiving emissions from the first and second reaction vessels, the first measurement device including a first photoelectric element and a first inlet optically connected with the first photoelectric element; and a connecting end arranging body that is operatively connected to the controller and that arranges and supports the second end of the first optical fiber and the fourth end of the second optical fiber along a first predetermined path; wherein the connecting end arranging body is driven by the controller along the first predetermined path by an arranging body Y axis transfer mechanism between: a first measurement position, in which the second end of the first optical fiber is optically connected with the first photoelectric element via the first inlet so that light based on an optical state within the first reaction vessel is transmittable from the first reaction vessel to the first photoelectric element, via at least the first optical fiber and the first inlet; and a second measurement position, in which the fourth end of the second optical fiber is optically connected with the first photoelectric element via the first inlet so that light based on an optical state within the second reaction vessel is transmittable from the second reaction vessel to the first photoelectric element, via at least the second optical fiber and the first inlet.
2. The optical measurement device of claim 1, further comprising: a first sealing lid coupled to the first nozzle, the first sealing lid having transparency; and a second sealing lid coupled to the second nozzle, the second sealing lid having transparency; wherein the first sealing lid is configured to be mounted on the first reaction vessel by detaching the first sealing lid from the first nozzle using a detaching mechanism; and wherein the second sealing lid is configured to be mounted on the second reaction vessel by detaching the second sealing lid from the second nozzle using the detaching mechanism.
3. The optical measurement device of claim 1, wherein the first reaction vessel is positioned within a first exclusive region of a vessel group in which two or more reaction vessels are arranged; wherein the first nozzle is configured to enter the first exclusive region; wherein a first liquid housing part is positioned within the first exclusive region and configured to house a first reaction solution for use in reactions; wherein a first sealing lid is configured to be transported to the first reaction vessel by the first nozzle to seal the first reaction solution in the first reaction vessel; wherein the second reaction vessel is positioned within a second exclusive region of the vessel group; wherein the second nozzle is configured to enter the second exclusive region; wherein a second liquid housing part is positioned within the second exclusive region and configured to house a second reaction solution for use in reactions; wherein a second sealing lid is configured to be transported to the second reaction vessel by the second nozzle to seal the second reaction solution in the second reaction vessel; and wherein the light guide plate of the light guide stage is configured to extend across the first and second exclusive regions so that: the first end of the first optical fiber enters the first exclusive region, and the third end of the second optical fiber enters the second exclusive region.
4. The optical measurement device of claim 3, further comprising: a third nozzle connected to the suction-discharge mechanism and configured to enter the first exclusive region; and a fourth nozzle connected to the suction-discharge mechanism and configured to enter the second exclusive region; wherein the third nozzle is configured to enter the first exclusive region together with the first nozzle; and wherein the fourth nozzle is configured to enter the second exclusive region together with the second nozzle.
5. The optical measurement device of claim 3, wherein the first and second exclusive regions of the vessel group are part of a first cartridge vessel having inspection information visibly displayed thereon so that images of the inspection information are configured to be obtained by a digital camera mounted on the nozzle head or the light guide stage.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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BEST MODE FOR CARRYING OUT THE INVENTION
(24) Next, an embodiment of the present invention is described with reference to the drawings. This embodiment is not to be interpreted as limiting the present invention unless particularly specified. Furthermore, in the embodiments or in the exemplary embodiments, the same objects are denoted by the same reference symbols, and the descriptions are omitted.
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(26) The optical measurement device for a reaction vessel 10 broadly has: a vessel group 20 in which a plurality (twelve in this example) of reaction vessel groups 23.sub.i (i=1, . . . , 12, omitted hereunder) are arranged; a nozzle head 50 that has a nozzle arranging portion 70 in which a plurality (twelve in this example) of nozzles 71.sub.i that detachably mount dispensing tips are arranged, and a light guide stage 32 that has a plurality (twelve in this example) of linking portions 31.sub.i provided with the ends of two or more light guide portions, which have a flexibility, that are directly or indirectly linkable with the apertures of the reaction vessels and optically connect to the linked reaction vessel interior; a measurement device 40 that is provided fixed to the nozzle head 50; a nozzle head transfer mechanism 51 that makes the nozzle head 50 movable in the X axis direction for example; a temperature controller 29 that performs predetermined temperature control with respect to the reaction vessel group 23.sub.i of the vessel group; a CPU+program 60 composed of a CPU, a ROM, a RAM, various types of external memory, communication functions such as a LAN, and a program stored in the ROM, and the like; and a control panel 13 having a display portion such as a liquid crystal display, and an operation portion, such as operation keys or a touch panel.
(27) The nozzle head 50 has: a stage Z axis transfer mechanism 35 that makes the light guide stage 32 movable in the Z axis direction with respect to the vessel group 20 independent of the nozzle arrangement portion 70; a nozzle Z axis transfer mechanism 75 that makes the nozzle arrangement portion 70 movable in the Z axis direction with respect to the vessel group 20 independent of the stage 30; a magnetic force part 57 that, by means of a magnet 571 provided on a narrow diameter portion 211ia of a dispensing tip 211i detachably mounted on the nozzle 71i such that it can approach and separate, is able to apply and remove a magnetic field with respect to the interior; a suction-discharge mechanism 53 that makes the suction and the discharge of liquids with respect to the dispensing tip 211i mounted on the nozzle 71i possible by performing the suction and the discharge of gases with respect to the nozzle 71i; and a punching mechanism 55 which is driven by the suction-discharge mechanism 53, for punching a film that covers the apertures of the liquid housing parts of the vessel group 20 to seal various liquids in advance. The stage transfer mechanism corresponds to the nozzle head transfer mechanism and the stage Z axis transfer mechanism.
(28) The nozzle head 50 further has a connecting end arranging body 30 that arranges and supports a plurality (twelve in this example) of connecting ends 34.sub.i, which are provided corresponding to the respective linking portions 31.sub.i and are provided with the back ends of optical fibers (bundle) 33.sub.i, which represent light guide portions in which the front ends thereof are provided to the linking portions 31.sub.i, such that, as an arranging surface, they are integrated along a predetermined path (a linear path along the Y axis direction in this example) provided on a vertical plane at a narrower spacing than the spacing between the linking portions 31.sub.i. Furthermore, the connecting end arranging body 30 is provided at a position that is separated from the light guide stage 32 and the reaction vessel group 23.sub.i.
(29) The measurement device 40 is able to respectively receive the light of specific wavelengths or specific wavelength bands of six types of fluorescent light, and also has six types of specific wavelength measurement devices 40.sub.j (j=1, . . . , 6, omitted hereunder) that are able to irradiate excitation light of six types of specific wavelengths or specific wavelength bands that are irradiated for the emission of light.
(30) The respective specific wavelength measurement devices 40.sub.j have measuring ends 44.sub.j that are provided approaching or making contact with the arrangement surface, and are successively connectable with the respective connecting ends 34.sub.i along the predetermined path (a linear path along the Y axis direction). Furthermore, the respective measuring ends 44.sub.j have two ends, namely a first measuring end 42.sub.j and a second measuring end 43.sub.j arranged along the Y axis direction. The first measuring ends 42.sub.j optically connect with an irradiation source provided to the specific wavelength measurement devices 40.sub.j. The second measuring ends 43.sub.j optically connect with a photoelectric device, such as a photomultiplier tube, provided to the specific wavelength measurement devices 40.sub.j.
(31) Furthermore, the nozzle head 50 has an arranging body Y axis transfer mechanism 41, which represents a light guide switching mechanism, that moves the connecting end arranging body 30 along the Y axis direction on the nozzle head 50 such that the respective connecting ends 34.sub.i arranged on the connecting end arranging body 30 and the respective measuring ends 44.sub.j are successively connected.
(32) Moreover, the light guide stage 32 has a heater 37, which represents the heating portion, for preventing condensation of the ends of the linking portions 31.sub.i or the mounted sealing lids 251.sub.i, which have transparency, by heating.
(33) The vessel group 20 comprises a plurality (twelve in this example) of exclusive regions 20.sub.i, in which one (in this example, one group corresponds to one) nozzle enters and the other nozzles do not enter, that correspond to the respective nozzles. The respective exclusive regions 20.sub.i have: a liquid housing part group 27.sub.i comprising a plurality of housing parts in which reagent solutions and the like are housed or are housable; a sealing lid housing part 25.sub.i in which one or two or more sealing lids 251.sub.i, which have transparency, that are detachably mounted on the nozzles are housed or are housable; and housing parts for tips and the like 21.sub.i that house, a plurality of dispensing tips 211.sub.i that are detachably mounted on the nozzles, and the samples, and the like. The liquid housing part group 27.sub.i has, at the very least, one or two or more liquid housing parts that house a magnetic particle suspension, and two or more liquid housing parts that house a solution for separating and extracting used for the separation and the extraction of nucleic acids or the fragments thereof. If necessary, it further has two or more liquid housing parts that house a solution for amplification used for the amplification of nucleic acids, and a liquid housing part that houses a sealing liquid for sealing the solution for amplification housed in a PCR tube 231.sub.i, which represents the reaction vessel, within the PCR tube 231.sub.i.
(34) It is preferable for the exclusive regions 20.sub.i to display a barcode as the sample information and the inspection information for identifying the exclusive regions 20.sub.i. Furthermore, the nozzle head 50 is provided with a single traversable nozzle 71.sub.0 in which liquids are transportable or dispensable by traversing (moving in the Y axis direction) the exclusive regions 20.sub.i, and suction and discharge is made to be performed by a traversable nozzle suction-discharge mechanism 17 that is separate from the suction-discharge mechanism 53. Consequently, the solution of DNA and the like housed in a given exclusive region 20.sub.i can be dispensed or delivered to the other exclusive regions 20.sub.k (k≠i). It is preferable for this movement in the Y axis direction to be also used by the arranging body Y axis transfer mechanism 41.
(35) The CPU+program 60 has, at the very least: a nucleic acid processing control portion 63 that performs instructions for a series of processes, such as extraction and amplification with respect to nucleic acids or the fragments thereof, and sealing of the solution for amplification, with respect to the temperature controller 29, the nozzle head transfer mechanism 51, the tip detaching mechanism 59, the suction-discharge mechanism 53, the magnetic force part 57, and the nozzle Z axis transfer mechanism 75; and a measurement control portion 61 that, after the nozzle head transfer mechanism 51 and the stage Z axis transfer mechanism 35 are controlled such that the linking portions 31.sub.i are simultaneously directly or indirectly linked with the apertures of the plurality (twelve in this example) of PCR tubes 231.sub.i, instructs a measurement by the measurement devices 40.sub.j by controlling the arranging body Y axis transfer mechanism 41 such that the optical fibers (bundle) 33.sub.i, which represent the light guide portions of the linking portions 31.sub.i, and the first measuring ends 42.sub.j and the second measuring ends 43.sub.j of the measuring ends 44.sub.j of the measurement devices 40.sub.j mentioned below are optically connected.
(36) Furthermore, the nucleic acid processing control portion 63 has an extraction control part 65 and a sealing lid control part 67. The nucleic acid processing control portion 63 has the extraction control part 65 that performs instructions with respect to the tip detaching mechanism 59, the suction-discharge mechanism 53, the magnetic force part 57, the nozzle Z axis transfer mechanism 75, the nozzle head transfer mechanism 51, and the stage Z axis transfer mechanism 35, for a series of processes with respect to the nucleic acids or the fragments thereof, and the sealing lid control part 67 that performs instructions with respect to the stage Z axis transfer mechanism 35 and the nozzle head transfer mechanism 51 for a sealing process by the sealing lids.
(37) Hereunder, a more specific first embodiment of the optical measurement device for a reaction vessel 10 mentioned above according to an embodiment of the present invention, is described with reference to
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(42) The cartridge vessel for PCR amplification 201.sub.1 has: the PCR tubes 231.sub.i, which represent the reaction vessel that are detachably linked with the twelve linking portions 31.sub.i provided to the light guide stage 32, via a single sealing lid 251.sub.i which has transparency; the liquid housing parts 271.sub.i which house a buffer solution necessary for the PCR reaction; the sealing lid housing parts 25.sub.i which house the sealing lids 251.sub.i; the housing part for tips and the like 21.sub.i that house the tips for punching for punching the film covering the PCR tubes 231.sub.i and the liquid housing parts 271.sub.i, and the dispensing tips 211.sub.i, and barcodes 81.sub.i that display the sample information and the inspection information relating to the cartridge vessels for PCR amplification 201.sub.i.
(43) The cartridge vessels for nucleic acid extraction 202.sub.i has: seven liquid housing parts 272.sub.i for example, that house various reagents for nucleic acid extraction; reaction vessels 232.sub.i that house the extracted nucleic acids; and barcodes 82.sub.i that display various information, such as the sample information and the inspection information, related to the cartridge vessel. The PCR tubes 231.sub.i and the reaction vessels 232.sub.i are temperature controllable by means of the temperature controller 29.
(44) The cartridge vessels for housing tips 203.sub.i has: a tip for punching that is able to punch the film covering the cartridge vessel for nucleic acid extraction 202.sub.i: two small-quantity dispensing tips that perform the dispensing of small quantities of liquids; housing parts for tips and the like 21.sub.i that house dispensing tips for separations that are able to perform separation by adsorbing magnetic particles on an inner wall by applying and removing a magnetic force from the exterior, and a barcode 83.sub.i that displays various information relating to the cartridge vessel 203.sub.i.
(45) The capacity of the PCR tube 231.sub.i, which represents the reaction vessel, is of the order of approximately 200 μL, and the capacity of the other reaction vessels, liquid housing parts, and tubes is of the order of approximately 2 mL.
(46) The PCR tube 231.sub.i is used for the amplification of nucleic acids or the fragments thereof, and temperature control is performed by means of the temperature controller 29 based on a predetermined amplification method, such as a thermal cycle (from 4° C. to 95° C.) for example. The PCR tube 231.sub.i is formed with two levels as shown in
(47) The liquid housing part group 272.sub.i houses the solutions for separating and extracting as follows. A first liquid housing part houses 40 μL of Lysis 1, a second liquid housing part houses 200 μL of Lysis 2, a third liquid housing part houses 500 μL of a binding buffer solution, a fourth liquid housing part houses a magnetic particle suspension, a fifth liquid housing part houses 700 μL of a washing liquid 1, a sixth liquid housing part houses 700 μL of a washing liquid 2, a seventh liquid housing part houses 50 μL of distilled water as a dissociation liquid, and an eighth liquid housing part, which is slightly separated, houses 1300 μL of isopropyl alcohol (isopropanol) used for the removal of protein and the like, as a portion of the solution for separating and extracting protein. The respective reagents and the like are prepacked as a result of the punchable film covering the respective apertures thereof.
(48) In addition, 1.2 mL of distilled water is housed in a separate distilled water reservoir, and tubes that house suspensions of bacteria, cells, and the like, or samples such as whole blood, are separately prepared for each of the respective exclusive regions 20.sub.i.
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(50) The nozzle head 50 is one having: a nozzle arranging portion 70 in which twelve nozzles 71.sub.i are arranged; a tip detaching mechanism 59 that is able to detach dispensing tips 211.sub.i mounted on the nozzles 71.sub.i; a suction-discharge mechanism 53; a magnetic force part 57 having twelve magnets 571 provided such that they are able to approach and separate with respect to the dispensing tips 211.sub.i; a light guide stage 32; twelve linking portions 31.sub.i provided to the light guide stage 32; a transfer mechanism portion 52 having a nozzle Z axis transfer mechanism 75 and a stage Z axis transfer mechanism 35; optical fibers (bundles) 33.sub.i representing flexible light guide portions that extend to the rear side from the linking portions 31.sub.i; a connecting end arranging body 30; the arranging body Y axis transfer mechanism 41; a measuring device 40 having a measuring end 44; a traversable nozzle 71.sub.0; and a suction-discharge mechanism 17 thereof.
(51) The nozzle arranging portion 70 is provided with a cylinder supporting member 73 that supports twelve cylinders 531.sub.i such that they are arranged along the Y axis direction at a predetermined pitch of 18 mm for example. The nozzles 71.sub.i are provided on the downward end of the cylinders 531.sub.i such that they are communicated with the cylinders 531.sub.i.
(52) The tip detaching mechanism 59 is provided with detaching shafts 593 on both sides, and has a tip detaching member 591 that detaches the twelve dispensing tips 211.sub.i from the nozzles 71.sub.i by sliding in the vertical direction.
(53) As shown specifically in
(54) As shown specifically in
(55) The magnetic force part 57 has a magnet 571 that is provided such that it can approach and separate with respect to the narrow diameter portions 211.sub.ia of the dispensing tips 211.sub.i detachably mounted on the nozzles 71.sub.i, and is able to apply and remove a magnetic field in the interior of the dispensing tips 211.sub.i.
(56) As shown specifically in
(57) As shown specifically in
(58) The stage Z axis transfer mechanism 35 has: a side plate 355 provided on the nozzle head substrate 753; a mount driving band-shaped member 354 that is supported by a timing belt 352 spanning between two sprockets 353 arranged in the vertical direction axially supported by the side plate 355, and vertically moves in the Z axis direction; and a motor attached to the rear side of the nozzle head substrate 753 that rotatingly drives the sprockets 353.
(59) As shown in
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(61) There are provided a connecting end arranging body 30 in which the connecting ends 34.sub.i provided corresponding to the respective linking portions 31.sub.i to which the front ends of the optical fibers (bundles) 33.sub.i which pass through the horizontal plate 32a of the light guide stage 32, are provided, and provided with the back ends thereof, are arranged on an arranging surface on a path along a straight line in the Y axis direction, which represents a predetermined path, at a shorter spacing than the spacing of the linking portions 31.sub.i; and six measuring ends that are provided in the vicinity of, or making contact with, the arranging surface, and are successively optically connectable with the connecting ends 34.sub.i along the straight line. There is also provided a measuring device 40 in which, by means of optical connections between the connecting ends and the measuring ends, the fluorescent light within the PCR tubes 231.sub.i, which represents the optical state, is receivable, and excitation light is also able to be irradiated.
(62) Furthermore, the light guide stage 32 has a cylinder-shaped body 311.sub.i, which retains the optical fibers (bundles) 33.sub.i extending to the rear side from the linking portions 31.sub.i such that they pass through the interior in order to prevent folding, protrudingly provided upward from the horizontal plate 32a directly above the linking portions 31.sub.i. In the same manner, the connecting end arranging body 30 is also provided with a cylinder-shaped body 301.sub.i, which retains the optical fibers (bundles) 33.sub.i extending from the connecting ends 34.sub.i such that they pass through the interior in order to prevent folding, on the connecting end 34.sub.i side.
(63) The arranging body Y axis transfer mechanism 41 that moves the connecting end arranging body 30 in the Y axis direction has: arms 412 and 413 provided to the connecting end arranging body 30; a joining body 411 that joins the arms 412 and 413 and the timing belt; a guide rail 414 that guides the Y axis movement of the joining body 411; and two sprockets spanned by the timing belt and arranged along the Y axis direction.
(64) The measuring device 40 is one that supports the measurement of fluorescent light and comprises six types of specific wavelength measuring devices 40.sub.j that are linearly aligned along a straight line in the Y axis direction, which represents the predetermined path, such that they support the measurement of six types of fluorescent light, and they are provided fixed on a substrate of the nozzle head 50, such as the frame that encloses the transfer mechanism portion 52, or a member that supports the same. Therefore, depending on the mechanism provided to the transfer mechanism portion 52, the measuring device 40 does not move.
(65) The measuring device 40 is one in which the measuring ends of the plurality of types (six in this example) of specific wavelength measuring devices 40.sub.j (j=1, 2, 3, 4, 5, 6), and therefore, in this case, the specific wavelength measuring devices 40.sub.j themselves are aligned in a single row form, and integrally fixed to a member joined with the nozzle head substrate 753 using fixtures 45.sub.j. The specific wavelength measuring devices 40.sub.j have: measuring ends 44.sub.j arranged along a straight line path in the Y axis direction which represents the predetermined path, such that they successively optically connect to the connecting ends 34.sub.i; light detectors 46.sub.j in which an optical system component having an irradiation source that irradiates excitation light to the PCR tubes 231.sub.i and a light receiving portion that receives the fluorescent light generated in the PCR tubes 231.sub.i are built-in; and circuit boards 47.sub.j. The measuring ends 44.sub.j have first measuring ends 42.sub.j that optically connect with the irradiation source, and second measuring ends 43.sub.j that optically connect with the light receiving portion. Here, the light detectors 46.sub.j and the circuit boards 47.sub.j correspond to the measuring device main body.
(66) The pitch between the respective connecting ends 34.sub.i, assuming a pitch between the linking portions 31.sub.i of 18 mm, is 9 mm, which is half thereof. Then, the pitch between the measuring ends 44.sub.j is 9 mm or less for example.
(67) There is a case where the first measuring ends 42.sub.j and the second measuring ends 43.sub.j of the measuring ends 44.sub.j of the respective specific wavelength measuring devices 40.sub.j are arranged aligned in a lateral direction (Y axis direction) along the straight line of the Y axis direction along the predetermined path, and a case where they are arranged aligned in a longitudinal direction (X axis direction). In the former case, without stopping the emission of the excitation light, the respective measuring devices successively receive light at a timing for receiving light determined based on the speed of the connecting end arranging body, the pitch between the connecting ends, the distance between the first measuring ends and the second measuring ends of the measuring ends, and the pitch between the measuring ends.
(68) On the other hand, in the latter case, as shown in
(69) The speed of the connecting end arranging body 30 with respect to the measuring ends 44.sub.j is determined with consideration of the stable light receivable time, the lifetime of the fluorescent light with respect to excitation light irradiation, the number of connecting ends, the pitch between the connecting ends, and the like (the distance of the predetermined path). In the case of a real-time PCR measurement, it is controlled such that it becomes 100 mm to 500 mm per second for example. In the present embodiment, since the movement is performed by sliding the arranging surface with respect to the measuring ends 44, the incidence of optical noise to the measuring ends 44 can be prevented. Furthermore, the connecting end arranging body 30 moves with respect to the measuring ends intermittently such that it momentarily stops at each pitch advance between the connecting ends or between the measuring ends, or continuously.
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(71) For the aperture of the wide-mouthed piping part 235i, since the sealing lid 251i that protrudes on the lower side of the sealing lid 251i, which has transparency, is mounted on the reaction vessel, it is mounted to the reaction vessel as a result of a tubular sealing portion 252i, which encloses the center portion of the sealing lid 251i in which light passes through, being fitted. At the time the sealing portion 252i is fitted, it is preferable for the diameter of the optical fibers (bundle) 33i, which represents the light guide portion that passes through the interior of the linking portion 31i, to be the same or larger than the diameter of the aperture of the narrow-mouthed piping part 233i. Consequently, it becomes possible to receive the light from the PCR tube 231i with certainty. The narrow-mouthed piping part 233i is housed within a block for temperature control that is heated or cooled by means of the temperature controller 29.
(72) In this example, the optical fibers (bundle) 33i comprise optical fibers (bundle) for irradiation 332i that are connectable with the second measuring end 43j and optical fibers (bundle) for receiving light 331i that are connectable with the first measuring end 42j.
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(74) It is preferable for a feeding device for samples and the like to be integrated to the optical measurement device for a reaction vessel 10. The feeding device for samples and the like is a device for dispensing and supplying parent samples and the like with respect to the vessel group 20, and the stage in which is integrated the vessel group 20 to which the parent samples and the like have been supplied, is automatically moved to the optical measurement device for a reaction vessel and the like. The feeding device for samples and the like, for example, has a parent vessel group that houses parent samples and the like, a tip detaching mechanism, a suction-discharge mechanism, and a single nozzle that, in addition to the suction and the discharge of gases being performed by means of the mechanisms, is detachably mounted with dispensing tips 211i. Furthermore, it has a nozzle head provided with a mechanism that moves along the Z axis direction with respect to the parent vessel group and the housing part group for tips and the like 21 of the vessel group 20, an X axis movable body provided with a Y axis transfer mechanism that moves the nozzle head in the Y axis direction with respect to the parent vessel group and the like, an X axis transfer mechanism that moves the X axis movable body along the X axis with respect to the parent vessel group and the like, and the parent vessel group. It is preferable for the parent vessel group to have a parent sample housing part group arranged in a 12 row×8 column matrix form that houses the parent samples to be supplied to the housing part group for tips and the like 21 of the vessel group 20, a distilled water and washing liquid group, and a reagent bottle group.
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(76) The specific wavelength measurement device 461 according to the present exemplary embodiment, in addition to having an optical fiber 469 for excitation light to be outgoing to the PCR tube 231i and an optical fiber 479 for light from the PCR tube 231i to be incoming, has a measuring end 441 provided on the lower ends of a first measuring end 421 of the optical fiber 469 and a second measuring end 431 of the optical fiber 479, an irradiation portion 462 that has a LED 467 that irradiates excitation light through the optical fiber 469 and a filter 468, and a light receiving portion that has an optical fiber 479, a drum lens 478, a filter 477, and a photodiode 472. This example shows a case where the first measuring end 421 and the second measuring end 431 are provided along a direction (X axis direction) perpendicular to the straight line of the Y axis direction, which represents the predetermined path.
(77) Next, a series of processing operations that perform real-time PCR of the nucleic acids of a sample containing bacteria using the optical measurement device for a reaction vessel 10 according to the embodiment is described. Step S1 to step S13 below correspond to separation and extraction processing.
(78) In step S1, the drawer 15 of the optical measurement device for a reaction vessel 10 shown in
(79) In step S2, following returning of the vessel group 20 and closing of the drawer 15, the start of the separation and extraction and amplification processing is instructed by means of the operation of the touch panel of the control panel 13 for example.
(80) In step S3, the extraction control part 65 provided to the nucleic acid processing controller 63 of the CPU+program 60 of the optical measurement device for a reaction vessel 10 instructs the nozzle head transfer mechanism 51 and moves the nozzle head 50 in the X axis direction, positions the tip for punching mounted to the nozzle 71.sub.i above the first liquid housing part of the liquid housing part group 27.sub.i of the vessel group, and punches the film covering the aperture of the liquid housing part by lowering the nozzle by means of the nozzle Z axis transfer mechanism 75, and in the same manner, the other liquid housing parts of the liquid housing part group 27.sub.i and the reaction vessel group 23.sub.i are successively punched by moving the nozzle head 50 in the X axis direction.
(81) In step S4, the nozzle head 50 is again moved in the X axis direction and moved to the housing part for tips and the like 21.sub.i, and the nozzles 71.sub.i are lowered by means of the nozzle Z axis transfer mechanism 75, and the dispensing tips 211.sub.i are mounted. Next, after being raised by the nozzle Z axis transfer mechanism 75, the dispensing tips 211.sub.i are moved along the X axis by means of the nozzle head transfer mechanism 51, and advanced to the eighth liquid housing part of the liquid housing part group 27.sub.i. Then a predetermined amount of isopropanol is aspirated from the liquid housing part, and they are again moved along the X axis direction, and predetermined amounts are respectively dispensed into the solution components (NaCl, SDS solutions) housed in the third liquid housing part and the fifth liquid housing part, and the distilled water housed in the sixth liquid housing part, so that 500 μL of a binding buffer solution (NaCl, SDS, isopropanol), 700 μL of a washing liquid 1 (NaCl, SDS, isopropanol), and 700 μL of a washing liquid 2 (water 50%, isopropanol 50%) are respectively prepared as solutions for separating and extracting within the third, the fifth, and the sixth liquid housing parts.
(82) In step S5, following movement to, among the housing parts for tips and the like 21.sub.i, the sample tube in which the sample is separately housed, the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i is loweringly inserted using the nozzle Z axis transfer mechanism 75, and, with respect to the suspension of the sample housed in the sample tube, following suspension of the sample within the liquid by repeating the suction and the discharge by raising and lowering the drive plate 536 of the suction-discharge mechanism 53, the sample suspension is aspirated within the dispensing tip 211.sub.i. The sample suspension is moved along the X axis by means of the nozzle head transfer mechanism 51 to the first liquid housing part of the liquid housing part group 27.sub.i housing the Lysis 1 (enzyme) representing the solution for separating and extracting, and the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i is inserted through the hole in the punched film, and the suction and the discharge is repeated in order to stir the sample suspension and the Lysis 1.
(83) In step S6, the entire amount of the stirred liquid is aspirated by the dispensing tip 211.sub.i, and incubation is performed by housing it in the reaction vessel 232.sub.i comprising the reaction tubes retained in the housing holes, that is set to 55° C. by means of the constant temperature controller. Consequently, the protein contained in the sample is broken down and made a low molecular weight. After a predetermined time has elapsed, the reaction mixture is left in the reaction tube, the dispensing tip 211.sub.i is moved to the second liquid housing part of the liquid housing part group 27.sub.i by means of the nozzle head transfer mechanism 51, and the entire amount of the liquid housed within the second liquid housing part is aspirated by using the nozzle Z axis transfer mechanism 75 and the suction-discharge mechanism 53, and it is transferred using the dispensing tip 211.sub.i by means of the nozzle head transfer mechanism 51, and the reaction solution is discharged within the third liquid housing part by penetrating the hole in the film and inserting the narrow diameter portion.
(84) In step S7, the binding buffer solution housed within the third liquid housing part, which represents a separation and extraction solution, and the reaction solution are stirred, the solubilized protein is further dehydrated, and the nucleic acids or the fragments thereof are dispersed within the solution.
(85) In step S8, using the dispensing tip 211.sub.i, the narrow diameter portion thereof is inserted into the third liquid housing part by passing through the hole in the film, the entire amount is aspirated and the dispensing tip 211.sub.i is raised by means of the nozzle Z axis transfer mechanism 75, and the reaction solution is transferred to the fourth liquid housing part, and the magnetic particle suspension housed within the fourth liquid housing part is stirred with the reaction solution. A cation structure in which Na+ ions bind to the hydroxyl groups formed on the surface of the magnetic particles contained within the magnetic particle suspension is formed. Consequently, the negatively charged DNA is captured by the magnetic particles.
(86) In step S9, the magnetic particles are adsorbed on the inner wall of the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i by approaching the magnet 571 of the magnetic force part 57 to the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i. In a state in which the magnetic particles are adsorbed on the inner wall of the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i, the dispensing tip 211.sub.i is raised by means of the nozzle Z axis transfer mechanism 75 and moved from the fourth liquid housing part to the fifth liquid housing part using the nozzle head transfer mechanism 51, and the narrow diameter portion 211.sub.ia is inserted by passing through the hole in the film.
(87) In a state in which the magnetic force within the narrow diameter portion 211.sub.ia is removed by separating the magnet 571 of the magnetic force part 57 from the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i, by repeating the suction and the discharge of the washing liquid 1 (NaCl, SDS, isopropanol) housed in the fifth liquid housing part, the magnetic particles are released from the inner wall, and the protein is washed by stirring within the washing liquid 1. Thereafter, in a state in which the magnetic particles are adsorbed on the inner wall of the narrow diameter portion 211.sub.ia as a result of approaching the magnet 571 of the magnetic force part 57 to the narrow diameter portion 211.sub.ia of the narrow diameter portion 211.sub.ia again, the dispensing tip 211.sub.i is, by means of the nozzle Z axis transfer mechanism 75, moved from the fifth liquid housing part to the sixth liquid housing part by means of the nozzle head transfer mechanism 51.
(88) In step S10, the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i is inserted by passing through the hole in the film using the nozzle Z axis transfer mechanism 75. By repeating the suction and the discharge of the washing liquid 2 (isopropanol) housed in the sixth liquid housing part in a state in which the magnetic force within the narrow diameter portion 211.sub.ia is removed by separating the magnet 571 of the magnetic force part 57 from the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i, the magnetic particles are stirred within the liquid, the NaCl and the SDS is removed, and the protein is washed. Thereafter, in a state in which the magnetic particles are adsorbed on the inner wall of the narrow diameter portion 211.sub.ia by approaching the magnet 571 of the magnetic force part 57 to the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i again, the dispensing tip 211.sub.i is, following raising by means of the nozzle Z axis transfer mechanism 75, moved from the sixth liquid housing part to the seventh liquid housing part in which the distilled water is housed, by means of the nozzle head transfer mechanism 51.
(89) In step S11, the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i is lowered through the hole by means of the nozzle Z axis transfer mechanism 75, and by repeating the suction and the discharge of the distilled water at a slow flow rate in a state where the magnetic force is applied within the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i, the washing liquid 2 (isopropanol) is substituted by water and is removed. Thereafter, by stirring the magnetic particles by repeating the suction and the discharge within the distilled water which represents the dissociation liquid, in a state in which the magnet 571 of the magnetic force part 57 is separated from the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i and the magnetic force is removed, the nucleic acids or the fragments thereof retained by the magnetic particles are dissociated (eluted) from the magnetic particles into the liquid. Thereafter, a magnetic field is applied within the narrow diameter portion and the magnetic particles are adsorbed on the inner wall by approaching the magnet 571 to the narrow diameter portion 211.sub.ia of the dispensing tip 211.sub.i, and the solution containing the extracted nucleic acids, and the like, is made to remain in the eighth liquid housing part. The dispensing tip 211.sub.i is moved to the storage part of the housing parts for tips and the like 21.sub.i in which the dispensing tip 211.sub.i was housed, by means of the nozzle head transfer mechanism 51, and the dispensing tip 211.sub.i to which magnetic particles are adsorbed, is detached from the nozzle 71.sub.i together with the magnetic particles and dropped into the storage part, using the detaching member 591 of the tip detaching mechanism 59.
(90) The following step S12 to step S15 corresponds to nucleic acid amplification and measurement processing.
(91) In step S12, a new dispensing tip 211.sub.i is mounted on the nozzle 71.sub.i, the solution housed within the eighth liquid housing part, which contains nucleic acids and the like, is aspirated, and by transferring it to the PCR tube 231.sub.i in which the solution for amplification 234.sub.i is housed beforehand, and discharging it, it is introduced into the vessel. As a result of moving the nozzle head 50 by means of the nozzle head transfer mechanism 51, the nozzle 71.sub.i is moved above the sealing lid housing part 25.sub.i of the vessel group 20, which houses the sealing lid 251.sub.i. Mounting is performed by lowering using the nozzle Z axis transfer mechanism 75 and fitting the indentation for linking 258.sub.i on the upper side of the sealing lid 251 to the lower end of the nozzle 71.sub.i. After being raised by the nozzle Z axis transfer mechanism 75, the sealing lid 251 is positioned above the PCR tube 231.sub.i using the nozzle head transfer mechanism 51, and by lowering the sealing lid 234.sub.i by means of the nozzle Z axis transfer mechanism 75, it is fitted with the aperture of the wide-mouthed piping part 235.sub.i of the PCR tube 231.sub.i, mountingly sealing it.
(92) In step S13, the nozzle head transfer mechanism 51 is instructed by means of an instruction from the measurement control portion 61, and by moving the nozzle head 50 along the X axis, the linking portion 31.sub.i of the light guide stage 32 is positioned above the PCR tube 231.sub.i, which is mounted with the sealing lid 251.sub.i. Then, by lowering the light guide stage 32 by means of the stage Z axis transfer mechanism 35, the linking portion 31.sub.i is inserted into the indentation of the sealing lid 251.sub.i, and the lower end thereof is made to make contact with, or adhere to, the bottom surface of the indentation.
(93) In step S14, due to an instruction by the nucleic acid processing controller 63, the temperature controller 29 instructs a temperature control cycle by real-time PCR, such as a cycle in which the PCR tube 231.sub.i is heated for five seconds at 96° C. and heated for 15 seconds at 60° C., to be repeated forty nine times for example.
(94) In step S15, when temperature control at each cycle by the nucleic acid processing controller 63 is started, the measurement control portion 61 determines the start of elongation reaction processing at each cycle, and instructs the continuous or intermittent movement of the connecting end arranging body 30 with respect to the measuring ends 44.sub.j of the measuring device 40. For the movement speed thereof, it is moved at a speed that is calculated based on the stable light receivable time, the fluorescence lifetime, and the number (twelve in this example) of exclusive regions 20.sub.i. Consequently, the receiving of light from all twelve PCR tubes 231.sub.i within the stable light receivable time becomes completed.
(95) In step S16, the measurement control portion 61 determines the moment of each optical connection between the optical fibers (bundles) 33.sub.i of the linking portions 31.sub.i and the first measuring end and the second measuring end of the measuring end 44, and instructs the receiving of light to the measuring device 40 for example.
(96) This measurement is executed with respect to cycles in which exponential amplification is performed, and an amplification curve is obtained based on the measurement, and various analyses are performed based on the amplification curve. At the time of the measurement, the measurement control portion 61 heats the heater 37 built into the light guide stage 32 and prevents the condensation on the sealing lid 251, and a clear measurement can be performed.
(97)
(98) As shown in
(99) There are no large differences with the first exemplary embodiment with respect to the section of the nozzle head 500 related to the dispensing device, which includes the nozzles, and the section related to the traversable nozzle, the transfer mechanisms of the nozzle head, and the arranging body transfer mechanism, and they are omitted from the descriptions. The nozzle head 500 has: a light guide stage 320; twelve linking portions 310.sub.i provided on the light guide stage 320; optical fibers (bundle) 33.sub.i that extend from the linking portions 310.sub.i on the rear side; a connecting end arranging body 300; a measuring device 400 having a measuring end comprising six types of specific wavelength measurement devices that are aligningly mounted on the light guide stage 320; and a sealing lid transport mechanism 125.
(100) The light guide stage 320 according to the second exemplary embodiment has a linking portion arranging body 322, in which two or more (twelve in this example) linking portions 310.sub.i that are simultaneously linkable with two or more (twelve in this example) reaction vessels 236.sub.i are arranged, that is movable in the horizontal direction (the X axis direction in this example) with respect to the light guide stage 320. Furthermore, by means of the movement of the linking portion arranging body 322, without moving the light guide stage 320, it is linkable with more reaction vessels 236.sub.i (three rows of reaction vessels with twelve per row in this example) than the number of reaction vessels (twelve in this example) that are simultaneously linkable by the linking portion arranging body 322.
(101) The light guide stage 320 has a horizontal plate 320a, a vertical plate 320b, and a triangular-shaped support side plate 320c. The horizontal plate 320a of the light guide stage 320, according to the arrangement of the linking portions 310.sub.i arranged on the linking portion arranging body 322, is etchingly provided with two or more, or twelve in this example, long holes 321.sub.i that correspond to shielding regions.
(102) The measurement device 400 is mounted fixed to the upper edge of the vertical plate 320b of the light guide stage 322. Therefore, since the light guide stage 320 is stationary at the time of receiving light, the measurement device 400 is immovably provided with respect to the reaction vessel and the light guide stage 320.
(103) The optical fibers (bundle) 33.sub.i having the end of the linking portion 310.sub.i, separate midway into optical fibers for receiving light (bundle) 331.sub.i and optical fibers for irradiation (bundle) 332.sub.i. The optical fibers for receiving light (bundle) 331.sub.i connect to a second connecting end 341.sub.i, and the optical fibers for irradiation (bundle) 332.sub.i connect to a first connecting end 342.sub.i, and are arranged as two paths along the Y axis direction on a downwardly facing horizontal plane, which represents an arranging surface on the lower side of the connecting end arranging body 300. At that time, the spacing between adjacent connecting ends on these respective paths is such that they are integrated at approximately half or one-third of the spacing of the linking portions for example. The first connecting ends 342.sub.i are successively connectable with the first measuring ends of the measurement device 400, and the second connecting ends 341.sub.i are successively connectable with the second measuring ends.
(104) As shown in
(105) The long holes 321.sub.i that are etchingly provided in the horizontal plate 320a reach the metallic plate 325. Holes 326 that are the same size as the apertures for light transmission are piercingly provided above the apertures of the reaction vessels 236.sub.i of the metallic plate 325 of the bottom of the long holes 321.sub.i, and are optically communicated with the bottom of the long holes 321.sub.i.
(106) The linking portions 310.sub.i provided on the linking portion arranging body and the front ends of the optical fibers (bundle) 33.sub.i provided in the interior are, as a result of approaching the sealing lids 253.sub.i, linked with the reaction vessels 236.sub.i.
(107)
(108) In
(109) The sealing lid 254.sub.i shown in
(110) The sealing lid 255.sub.i shown in
(111) The sealing lid 256.sub.i shown in
(112) The sealing lid 257.sub.i shown in
(113)
(114) The sealing lid transporting body 125 is one having: a prismatic substrate 128 that is movable in the X axis direction with respect to the vessel group 20, which has at least three rows of reaction vessels 236.sub.i of twelve per row; one or two or more (twelve in this example) grippers 127.sub.i arranged on the prismatic substrate 128 according to the arrangement of the reaction vessels that grip the cover plate such that, with respect to the sealing lid 253.sub.i (to 256.sub.i), the lower side is exposed in a state in which the mounting portion is mountable to the reaction vessel; and a bottom plate 126 that is mounted on the lower side of the prismatic substrate 128.
(115) As shown in the cross-sectional view of
(116) Next, the processing operation using the nozzle head 500 according to the second exemplary embodiment is described.
(117) Among the processes described in the first exemplary embodiment, the separation and extraction process is omitted, and step S′12 to step S′16, which correspond to nucleic amplification and measurement processes, are described.
(118) In step S′12, a new dispensing tip 211.sub.i is mounted on the nozzle 71.sub.i, the solution containing nucleic acids and the like, which is housed within the eighth liquid housing part is aspirated, transported to the reaction vessel 236.sub.i in which the solution for amplification 234.sub.i is housed beforehand, and discharged and introduced into the vessel. As a result of moving the nozzle head 500 by means of the nozzle head transfer mechanism 51, the sealing lids 253.sub.i from the sealing lid housing part in the sealing lid transporting body 125 in which twelve sealing lids 253.sub.i are housed, are simultaneously housed in the cavity 124.sub.i of the grippers 127.sub.i, and gripped.
(119) Since the sealing lid transporting body 125 gripping the sealing lid 253.sub.i is linked with the light guide stage 320, then by using the stage Z axis transfer mechanism 35 and moving it somewhat upwardly together with the stage 320 and then moving it in the X axis direction, and by transporting it to above the reaction vessels 236.sub.i and lowering it, the twelve sealing lids 253.sub.i are sealed by mounting the clamps 253.sub.ib, which are exposed on the lower side from the sealing lid transporting body 125, to the PCR tubes 236.sub.i. In the same manner, the rows of the twenty four reaction vessels of the other two rows are successively sealed by the sealing lids.
(120) In step S′13, due to an instruction by the measurement control portion 61, as a result of instructing the nozzle head transfer mechanism 51 and moving the nozzle head 500 along the X axis, the light guide stage 320 is moved such that it covers the thirty six reaction vessels of the three rows, on which the sealing lids are mounted.
(121) In step S′14, due to an instruction by the nucleic acid processing control portion 63, the temperature controller 29 instructs a temperature control cycle by real-time PCR, such as a cycle in which the PCR tubes 231.sub.i are heated for five seconds at 96° C. and heated for fifteen seconds at 60° C., to be repeated forty nine times for example.
(122) In step S′15, when temperature control at each cycle is started by the nucleic acid processing control portion 63, the measurement control portion 61 determines the start of the elongation reaction process at each cycle, moves the linking portion arranging body 322 provided on the light guide stage 320 over the light guide stage 320, indirectly links the respective linking portions 310.sub.i that are inserted into the long holes 321.sub.i provided on the light guide stage 320 via the reaction vessels and the sealing lids 253.sub.i, and successively receives the light from the reaction vessels while irradiating excitation light from the measurement device to the interior of the reaction vessels. At the same time, the continuous or intermittent movement of the connecting end arranging body 300 with respect to the respective measuring ends 44.sub.j of the measurement device 400 is instructed. The movement speed thereof is such that movement is performed at a speed that is calculated based on the stable light receivable time, the fluorescence lifetime, the number (three rows of twelve reaction vessels per row in this example) of reaction vessels of the exclusive regions 20.sub.i that are measurable by the light guide stage 320, and the like. Consequently, by moving the linking portion arranging body 322 over the light guide stage 320 within the stable light receivable time, in this example, measurements can be performed in parallel with respect to thirty six reaction vessels of three rows, with twelve per row.
(123) In step S′16, the measurement control portion 61 determines the moment of the respective optical connections between the optical fibers (bundle) of the linking portions 310.sub.i and the first measuring end and the second measuring end of the measuring end 44, and instructs the irradiation of excitation light and the receiving of light to the measurement device 400.
(124)
(125)
(126) The optical measurement device for a reaction vessel 110 according to the second embodiment differs from the optical measurement device for a reaction vessel 10 according to the first embodiment in the aspect that the nozzle head 150 thereof has a light guide stage 132 that is different from the light guide stage 32. The light guide stage 132 according to the second embodiment differs from the light guide stage 32 according to the first embodiment in the aspects that it has a plurality (twelve in this example) of linking portions 131.sub.i in which the front ends of optical fibers, which represent two or more light guide portions, which have a flexibility, that optically connect with the interior of the PCR tubes 231.sub.i, and an optical element for collecting light are provided in the interior, and the heat source of the heater 137, which represents a heating portion for heating the reaction vessels, is provided not to the light guide stage 132, but to the vessel group 120 or the stage.
(127) Further, it differs in the aspects that the sealing lids 251.sub.i are transported not by the nozzles 71.sub.i but by fitting to the linking portions 131.sub.i, and are detached from the linking portions by means of a dedicated sealing lid detaching mechanism 39. Therefore, the sealing lid control portion 167, and therefore, the nucleic acid processing control portion 163 and the CPU+program 160 differ from the device 10 according to the first embodiment.
(128) The vessel group 120 is one in which twelve exclusive regions 120.sub.i (i=1, . . . , 12), wherein the longitudinal direction thereof is along the X axis direction and housing parts are arranged in a single row form, are arranged in the Y axis direction for example. The respective exclusive regions 120.sub.i have: a reaction vessel group 23.sub.i; a liquid housing part group 27.sub.i; a sealing lid housing part 25.sub.i that houses sealing lids 251.sub.i, which have transparency, that are detachably mounted on the linking portions 131.sub.i provided to the light guide stage 132; and housing parts for tips and the like 21.sub.i.
(129) The reaction vessel 23.sub.i, the temperature controller 29, and the heater 137 correspond to the reaction vessel control system 90.
(130)
(131) Here, since the diameter of the linking portion 131.sub.i is thicker than the nozzle 71.sub.i, the sealing lid 251.sub.i to be mounted on the PCR tube is transported by the linking portion 131.sub.i. Consequently, by utilizing the transfer mechanism of the magnet 571 of the magnetic force part 57, a sealing lid detaching mechanism 39 is provided that has a comb-shaped detaching member 391 in which a notch portion, which has a semicircular-shaped arch that is approximately equivalent to the diameter of the twelve linking cylinders provided such that they can approach and separate with respect to the linking portion 131.sub.i, is arranged. In the present exemplary embodiment, since the detachment of the sealing lids can be performed by utilizing existing mechanisms, the expansion of the device scale, and increases in complexity, can be prevented.
(132)
(133) As shown in
(134) As shown in
(135) The temperature controller 291 for PCR has: a block for temperature control 292.sub.i that makes contact with, and is housable in, the narrow-mouthed piping part 233.sub.i of the PCR tube 231.sub.i, which represents the reaction vessel; a Peltier element 293.sub.i; and a heat sink 294.sub.i.
(136) The narrow-mouthed piping part 233.sub.i of the PCR tube 231.sub.i has a lower side wall section 233a.sub.i of the section in which the block for PCR 292.sub.i is making contact and is provided. Furthermore, it has an upper side wall section 235a.sub.i provided on the upper side leaving a spacing with the lower side wall section 233a.sub.i that corresponds to the wall section of the wide-mouthed piping part 235.sub.i that makes contact with the block for heating 137c of the heater.
(137) According to the present exemplary embodiment, firstly, by means of an instruction by the sealing lid control portion 167 (the CPU+program 160), the nozzle head transfer mechanism 51 is instructed and following movement of the respective linking portions 131.sub.i of the light guide stage 132 to the sealing lid housing parts 25.sub.i, the stage Z axis transfer mechanism 35 is instructed and the sealing lids 251.sub.i are fitted and mounted to the linking portions 131.sub.i. Next, by fitting the apertures of the predetermined PCR tubes 231.sub.i with the sealing lids 251.sub.i, the linking portions 131.sub.i are simultaneously linked with the PCR tubes 231.sub.i.
(138) Next, according to the temperature control by the temperature controller 29 as a result of an instruction by the measurement control portion 161, in the case of PCR, by controlling the heater 137 such that the upper side wall section 233b.sub.i is heated at a fixed temperature (100° C. for example) that is several degrees, or preferably approximately 5° C., higher than the maximum predetermined temperature (94° C. for example), the sealing lid 251.sub.i fitted to the wide-mouthed piping part 235.sub.i of the PCR tube 231.sub.i is heated, and condensation of the sealing lid can be prevented. At that time, the upper side wall section 235a.sub.i is separated by a fixed spacing from the lower side wall section 233a.sub.i, in which temperature control is performed, and the upper side wall section 233a.sub.i, which has a smaller surface area than the lower side wall section, is heated by bringing the heat source into contact or into its vicinity. Consequently, the effect of heating the upper side walls section 235a.sub.i is such that the lower surface of the sealing lid 251.sub.i, which is provided at a position near the upper side walls section 235a.sub.i, is heated, and condensation can be prevented.
(139) On the other hand, since the linking portion 131.sub.i is only making contact with the upper side of the sealing lid 251.sub.i via the circular edge portion 131d.sub.i, the effect of heating is not as much as with respect to the sealing lid 251.sub.i. In the same manner, the lower side wall section 233a.sub.i is temperature controlled to the predetermined temperature using a Peltier element having a heating and cooling function, and furthermore, measurements are simultaneously performed. After completion of the measurement, then by means of an instruction by the sealing lid control portion 167, the linking portion 131.sub.i is made to approach using the detaching member 391, and then by upwardly moving the light guide stage 132 by means of the stage Z axis transfer mechanism 35, the sealing lid 251.sub.i is detached from the linking portion and while remaining on the PCR tube 231.sub.i, the linking portion is moved and the linking is released.
(140)
(141)
(142) The foregoing exemplary embodiments have been specifically described in order to better understand the present invention, and they are in no way limiting of other embodiments. Therefore, modifications are possible within a scope that does not depart from the gist of the invention. The configurations, shapes, materials, arrangements, and amounts of the nozzles, the dispensing tips, the punching tips, the vessel group, the exclusive regions thereof, the housing parts, the measuring ends, the measurement devices, the specific wavelength measurement devices, the suction-discharge mechanism, the transfer mechanism portion, the magnetic force part, the heating portion, the reaction vessels, the sealing lids, the light guide stage, the linking portions, the light guide portions, the connecting ends, the connecting end arranging body, the linking portion arranging body, the nozzle head, the temperature controller, the nozzle detaching mechanism, and the sealing lid detaching mechanism, and the like, and the utilized reagents and samples are also in no way limited by the examples illustrated in the exemplary embodiments. Furthermore, although the nozzles were made to move with respect to the vessel group, it is possible to also move the vessel group with respect to the nozzles.
(143) Furthermore, in the foregoing descriptions, although the amplification solution was sealed using a sealing lid for the sealing of the reaction vessel for PCR, it may be made such that, in its place or in combination, it is sealed using a sealing liquid, such as mineral oil. Furthermore, in place of punching by mounting a tip for punching on the nozzles, it is possible to use a punching pin that is driven by the suction-discharge mechanism. Moreover, although a real-time PCR measurement was described in the foregoing descriptions, it is in no way limited to this measurement, and it can be applied to other various measurements in which temperature control is performed. In the foregoing descriptions, although a case where the measurement device is provided to a dispensing device was described, it is not necessarily limited to this. Although only an optical system using optical fibers was described, it is possible to also employ an optical system using a lens system in the interior of the measurement device.
(144) Furthermore, the devices described in the respective exemplary embodiments of the present invention, the components that form these devices, or the components that form these components, can be appropriately selected, and can be mutually combined by applying appropriate modifications. The spatial representations within the present application, such as “above”, “below”, “interior”, “exterior”, “X axis”, “Y axis”, and “Z axis” are for illustration only, and are in no way limiting of the specific spatial directions or arrangements of the construction.
INDUSTRIAL APPLICABILITY
(145) The present invention is related to fields in which the processing, testing, and analysis of nucleic acids, which primarily includes DNA, RNA, mRNA, rRNA, and tRNA for example, is required, and is related to industrial fields, agricultural fields such as food, agricultural products, and fishery processing, chemical fields, pharmaceutical fields, health care fields such as hygiene, insurance, diseases, and genetics, and scientific fields such as biochemistry or biology for example. The present invention is, in particular, able to be used in processing and analysis that handles various nucleic acids, and the like, such as PCR and real-time PCR.
BRIEF DESCRIPTION OF THE REFERENCE SYMBOLS
(146) 10, 110 Optical measurement device for reaction vessel 20, 120 Vessel group 20.sub.i, 120.sub.i (.sub.i=1, . . . , 12) Exclusive regions 211.sub.i (.sub.i=1, . . . , 12) Dispensing tips 231.sub.i, 236.sub.i (.sub.i=1, . . . , 12) PCR tubes (reaction vessels) 30, 300 Connecting end arranging body 31.sub.i, 131.sub.i (.sub.i=1, . . . , 12) Linking portions 32, 320, 132 Light guide stage 33.sub.i Optical fibers (light guide portions) 40, 400 Measurement device 40.sub.j (.sub.j=1, . . . , 6) Specific wavelength measurement devices 44 Measuring end 50, 500, 150 Nozzle head 52 Transfer mechanism portion 53 Suction-discharge mechanism 59 Tip detaching mechanism 61, 161 Measurement control portion 70 Nozzle arranging portion 71.sub.i (.sub.i=1, . . . , 12) Nozzle