Dispensing cylinder, and dispensing device and dispensing treatment method using same

Abstract

A dispensing cylinder capable of performing a dispensing treatment for both minute and large amounts of liquid includes: a cylinder having a cavity; a nozzle provided at one end of the cylinder, having a through hole in fluid communication with the cavity and extending in an axial direction of the cavity, and a plunger slidable in the cavity in the axial direction. The cavity has a large diameter region having a large inner peripheral surface and a small diameter region having a second inner peripheral surface. The plunger has a thick shaft portion slidable in the large diameter region and a thin shaft portion protruding from a distal end of the thick shaft portion in the axial direction and slidable in the small diameter region.

Claims

1. A dispensing cylinder comprising: a cylinder having a cavity that extends through the cylinder in an axial direction, the cavity having a large diameter region with a large inner peripheral surface, a small diameter region having a small inner peripheral surface, and a floating region located between the large diameter region and the small diameter region, wherein a diameter of the large inner peripheral surface is larger than a diameter of the small inner peripheral surface; a nozzle configured to be coupled to a dispensing tip and provided at one end of the cylinder, the nozzle having a through hole that is fluidly coupled to the cavity and extends in the axial direction; and a plunger provided within the cavity and configured to slide in the axial direction, the plunger having a thick shaft portion, a thin shaft portion protruding from a distal end of the thick shaft portion in the axial direction and terminating in a distal end, and a guide portion extending from the distal end of the thin shaft portion in the axial direction, wherein a diameter of the thick shaft portion is larger than a diameter of the thin shaft portion; wherein the thick shaft portion is configured to slide in the large diameter region and the floating region and wherein the thin shaft portion is configured to slide in the small diameter region; and wherein the guide portion has a ventilation passage configured to fluidly couple the through hole of the nozzle with the floating region.

2. The dispensing cylinder according to claim 1, further comprising: a first airtight seal member coupled to the large inner peripheral surface of the large diameter region or an outer peripheral surface of the thick shaft portion; and a second airtight seal member coupled to the small inner peripheral surface of the small diameter region or an outer peripheral surface of the thin shaft portion.

3. The dispensing cylinder according to claim 1, wherein at least a portion of the ventilation passage extends in the axial direction.

4. The dispensing cylinder according to claim 3, wherein the ventilation passage further comprises a lateral hole portion.

5. A dispensing device, comprising: a stage; a container group disposed on the stage and including a liquid-storing unit capable of storing various reagents and specimens; one or more dispensing tips; a tip-storing unit for storing the one or more dispensing tips; one or more dispensing cylinders, each of the one or more dispensing cylinders comprising: a cylinder having a cavity that extends through the cylinder in an axial direction, the cavity having a large diameter region with a large inner peripheral surface, a small diameter region having a small inner peripheral surface, and a floating region located between the large diameter region and the small diameter region, wherein a diameter of the large inner peripheral surface is larger than a diameter of the small inner peripheral surface; a nozzle configured to be coupled to a dispensing tip and provided at one end of the cylinder, the nozzle having a through hole that is fluidly coupled to the cavity and extends in the axial direction; and a plunger provided within the cavity and configured to slide in the axial direction, the plunger having a thick shaft portion, a thin shaft portion protruding from a distal end of the thick shaft portion in the axial direction and terminating in a distal end, and a guide portion extending from the distal end of the thin shaft portion in the axial direction, wherein a diameter of the thick shaft portion is larger than a diameter of the thin shaft portion; wherein the thick shaft portion is configured to slide in the large diameter region and the floating region, and wherein the thin shaft portion is configured to slide in the small diameter region; and wherein the guide portion has a ventilation passage configured to fluidly couple the through hole of the nozzle with the floating region; wherein each of the one or more dispensing tips is detachably attached to a respective nozzle of each dispensing cylinder; a dispensing head comprising a nozzle arrangement unit in which the one or more nozzles of the one or more dispensing cylinders are arranged; a suction/discharge driving unit for moving respective one or more of the plungers of the one or more dispensing cylinders in a vertical direction and sucking and discharging liquid with respect to the one or more dispensing tips; and a nozzle moving mechanism capable of moving the one or more nozzles relative to the container group; and a minute amount/large amount judgement and instruction unit coupled to the suction/discharge driving unit and configured to: receive an instruction to suck and discharge a predetermined amount of liquid to the one or more dispensing tips; determine whether the predetermined amount is within a first amount range or a second amount range, wherein the first amount range is separate from and smaller than the second amount range, responsive to a determination that the predetermined amount is within the first amount range, instruct the suction/discharge driving unit to slide the thin shaft portions of the one or more dispensing cylinders by a distance corresponding to the predetermined amount within the small diameter region, and responsive to a determination that the predetermined amount is within the second amount range, instruct the suction/discharge driving unit to slide the thick shaft portions of the one or more dispensing cylinders by a distance corresponding to the predetermined amount within the large diameter region.

6. The dispensing device according to claim 5, further comprising: a first airtight seal member coupled to an inner peripheral surface of the large diameter region or an outer peripheral surface of the thick shaft portion; and a second airtight seal member coupled to an inner peripheral surface of the small diameter region or an outer peripheral surface of the thin shaft portion.

7. The dispensing device according to claim 5, wherein at least a portion of the ventilation passage extends in the axial direction.

8. The dispensing device according to claim 5, wherein: the dispensing head further comprises a tip detaching mechanism configured to detach each of the one or more dispensing tips attached to the one or more nozzles of the one or more dispensing cylinders responsive by lowering the suction/discharge driving unit.

9. A dispensing treatment method, comprising: receiving, at a control unit, an instruction to suck or discharge a predetermined amount of liquid; determining, at a minute amount/large amount judgement and instruction unit within the control unit, whether the predetermined amount is within a first amount range or a second amount range, wherein the first amount range is separate from and smaller than the second amount range; attaching, using a dispensing head moving mechanism operably coupled to the control unit, a dispensing tip to a dispensing cylinder, wherein the dispensing cylinder comprises: a cylinder having a cavity that extends through the cylinder in an axial direction, the cavity having a large diameter region with a large inner peripheral surface, a small diameter region having a small inner peripheral surface, and a floating region located between the large diameter region and the small diameter region, wherein a diameter of the large inner peripheral surface is larger than a diameter of the small inner peripheral surface; a nozzle configured to be coupled to the dispensing tip and provided at one end of the cylinder, the nozzle having a through hole that is fluidly coupled to the cavity and extends in the axial direction; and a plunger provided within the cavity and configured to slide in the axial direction, the plunger having a thick shaft portion, a thin shaft portion protruding from a distal end of the thick shaft portion in the axial direction and terminating in a distal end, and a guide portion extending from the distal end of the thin shaft portion in the axial direction, wherein a diameter of the thick shaft portion is larger than a diameter of the thin shaft portion; wherein the thick shaft portion is configured to slide in the large diameter region and the floating region, and wherein the thin shaft portion is configured to slide in the small diameter region; and wherein the guide portion has a ventilation passage configured to fluidly couple the through hole of the nozzle with the floating region; responsive to a determination that the predetermined amount is within the first amount range, sucking and discharging the first amount range of liquid to the dispensing tip by sliding the thin shaft portion in the small diameter region by a distance corresponding to the predetermined amount; and responsive to a determination that the predetermined amount is within the second amount range, sucking and discharging the second amount range of liquid to the dispensing tip by sliding the thick shaft portion in the large diameter region by a distance corresponding to the predetermined amount.

10. The dispensing treatment method according to claim 9, wherein: sucking and discharging the first amount range of liquid further comprises the thick shaft portion moving in the floating region.

11. The dispensing treatment method according to claim 9, wherein at least a portion of the ventilation passage extends in the axial direction.

Description

BRIEF DESCRIPTION OF DRAWINGS

(1) FIG. 1 is a perspective view illustrating a dispensing cylinder according to a first embodiment of the present invention and a cross-sectional perspective view illustrating the inside thereof, obtained by cutting the dispensing cylinder.

(2) FIG. 2 is an operation explanatory view of the dispensing cylinder according to the first embodiment of the present invention.

(3) FIG. 3 is a perspective view illustrating a dispensing cylinder according to a second embodiment of the present invention and a cross-sectional perspective view illustrating the inside thereof, obtained by cutting the dispensing cylinder.

(4) FIG. 4 is an operation explanatory view of the dispensing cylinder according to the second embodiment of the present invention.

(5) FIG. 5 is a block diagram illustrating a dispensing device according to the first or second embodiment of the present invention.

(6) FIG. 6 is a perspective view illustrating the dispensing device according to the first or second embodiment of the present invention.

(7) FIG. 7 is an enlarged perspective view illustrating the dispensing device of FIG. 6 with a large amount dispensing tip attached to a nozzle.

(8) FIG. 8 is an enlarged perspective view illustrating the dispensing device of FIG. 6 with a minute amount dispensing tip attached to a nozzle.

(9) FIG. 9 is a plan view illustrating a container group disposed on a stage of the dispensing device illustrated in each of FIGS. 6 to 8.

DESCRIPTION OF EMBODIMENTS

(10) Subsequently, embodiments of the present invention will be described with reference to the drawings. Note that the embodiments should not be construed as limiting the present invention unless otherwise specified. In the embodiments or embodiment examples, the same reference numeral is used to denote the same part, and descriμLion thereof is omitted.

(11) A dispensing cylinder 10 according to a first embodiment of the present invention will be described with reference to FIGS. 1 and 2.

(12) The dispensing cylinder 10 according to the present embodiment includes: a nozzle 13 provided at one end of a cylinder 11 having a cavity (18, 19, 16) therein, having a nozzle suction/discharge port 13a at a distal end thereof, having a through hole 13b communicating with the cavity (18, 19, 16) and extending in the axial direction of the cavity, and having an attachment portion 13c capable of attaching a dispensing tip (211, 212) to the outside thereof; and a plunger 12 provided so as to be slidable in the cavity (18, 19, 16) in the axial direction and having a flange 12a to be connected to a driving unit such as a stepping motor at an outer side end portion of the cylinder 11. The nozzle 13 is formed separately from the cylinder 11 and attached to the one end via a connecting portion 11b.

(13) The cavity (16, 18, 19) has a large diameter region 18 having a large diameter inner peripheral surface and a small diameter region 16 provided on the side of the nozzle 13 with respect to the large diameter region and having a small diameter inner peripheral surface. Here, in the present embodiment, the inner diameter of the through hole 13b is larger than the inner diameter of the small diameter region 16, but smaller than the inner diameter of the large diameter region 18.

(14) The plunger 12 includes: a rod 12b penetrating an opening 11a provided at the other end of the cylinder 11 and provided in the cavity (16, 18, 19) of the cylinder 11 in the axial direction; a thick shaft portion 15 provided coaxially with the rod 12b, formed in a columnar shape having an outer diameter larger than the outer diameter of the rod 12b, and provided so as to be slidable in the large diameter region 18; and a thin shaft portion 14 protruding from a distal end surface of the thick shaft portion 15 or the rod 12b in the axial direction and provided so as to be slidable in the small diameter region 16. A length (d3) of the thin shaft portion 14 in the axial direction is shorter than the stroke (D) of the thick shaft portion 15.

(15) Furthermore, in the dispensing cylinder 10 according to the present embodiment, an O-ring 17a as an airtight seal member is provided so as to surround an axis in a peripheral direction on an inner peripheral surface of an upper end portion (the position is almost d5=0) of the small diameter region 16, and an O-ring 17b as another airtight seal member is provided so as to surround the axis in a peripheral direction on an outer peripheral surface of the thick shaft portion 15 in the thick shaft portion 15 (a distance to the airtight seal member is d1).

(16) Furthermore, in the dispensing cylinder 10 according to the present embodiment, between the large diameter region 18 and the small diameter region 16, a floating region 19 in which the thick shaft portion 15 is floatable in the axial direction is provided coaxially with the large diameter region 18 and the small diameter region 16. A length (d0) of the floating region 19 is equal to or longer than the sum of the length (d3) of the thin shaft portion 14 in the axial direction and the length (d1) of the thick shaft portion. In the present embodiment, the floating region 19 has an inner peripheral surface on which the thick shaft portion 15 does not slide, and has an inner diameter with a maximum diameter larger than the large diameter. Therefore, on a boundary with the large diameter region 18, a reversely tapered inclined surface is formed toward the nozzle 13. On a boundary with the small diameter region 16, at least one step projecting inward is formed toward a distal end of the nozzle so as to partition an inner wall surface of the cavity.

(17) FIG. 2 illustrates operation of the dispensing cylinder 10 according to the present embodiment. FIGS. 2(1) and 2(2) illustrate suction operation of a minute amount of liquid. In FIG. 2(1), the thick shaft portion 15 is located at a bottom dead center of the plunger 12 as a lowermost end of the floating region 19, that is, an upper end of the small diameter region 16. Therefore, the thin shaft portion 14 is inserted into the small diameter region 16. In this state, a distal end of a dispensing tip attached to the nozzle 13 is inserted into a container-storing liquid.

(18) Note that the stroke of the thick shaft portion 15 is represented by D, a distance from the lowermost end of a distal end of the thick shaft portion 15 in the axial direction is represented by d, the length of the thin shaft portion 14 is represented by d3, and a distance from the distal end of the thick shaft portion 15 to the airtight seal member 17b is represented by d1. Therefore, the length d0 of the floating region 19 is equal to or larger than d3+d1. If the sum of the lengths of the small diameter region and the through hole of the nozzle is represented by d2, as described above, 0≤d≤D, d3<d2, and d3<D are satisfied.

(19) In FIG. 2(2), when the plunger 12 is raised from the bottom dead center by a distance d (<d3), the thin shaft portion 14 slides in the small diameter region 16 and rises by the distance d, the thick shaft portion 15 moves by d in the floating region 19, and liquid flows into the dispensing tip attached to the nozzle 13 in an amount of S2×d.

(20) d<d3 in FIGS. 2(1) and 2(2) corresponds to the minute amount suction and discharge section.

(21) In FIG. 2(3), when the plunger 12 is moved from the bottom dead center by a distance d=d3, the thin shaft portion 14 starts to be withdrawn from the small diameter region 16, the thick shaft portion 15 enters the large diameter region 18, and suction of gas passing through an opening 13a of the nozzle 13 into the floating region 19 and the large diameter region 18 thereby starts.

(22) In FIG. 2(4), when the plunger 12 is moved from the bottom dead center by a distance d (>d3), the thin shaft portion 14 moves in the floating region 19, then the thick shaft portion 15 slides in the large diameter region 18, and liquid is sucked into the dispensing tip attached to the nozzle 13 in an amount of S1×{d−(d0−d1)} generally (in the above case (2)), and in an amount of S1×(d−d3) in the case of FIG. 2. Here, S1 and S2 are cross-sectional areas of the large diameter region 18 and the small diameter region 16 cut perpendicularly to the axial direction, respectively.

(23) Subsequently, a dispensing cylinder 10 according to a second embodiment of the present invention will be described with reference to FIGS. 3 and 4.

(24) The dispensing cylinder 30 according to the present embodiment includes: a nozzle 33 provided at one end of a cylinder 31 having a cavity (36, 38, 39) therein, having a nozzle suction/discharge port 33a at a distal end thereof, having a through hole 33b communicating with the cavity (36, 38, 39) and extending in the axial direction of the cavity, and having an attachment portion 33c capable of attaching a dispensing tip (211, 212) to the outside thereof; and a plunger 32 provided so as to be slidable in the cavity (36, 38, 39) in the axial direction and having a flange 32a to be connected to a driving unit such as a stepping motor at an outer side end portion of the cylinder 31. The nozzle 33 is formed separately from the cylinder 31 and attached to the one end via a connecting portion 31b.

(25) The cavity (36, 38, 39) has a large diameter region 38 having a large diameter inner peripheral surface and a small diameter region 36 provided on the side of the nozzle 33 with respect to the large diameter region 38 and having a small diameter inner peripheral surface. Here, in the present embodiment, the inner diameter of the through hole 33b is larger than the inner diameter of the small diameter region 36, but smaller than the inner diameter of the large diameter region 38.

(26) The plunger 32 includes: a rod 32b penetrating an opening 31a provided at the other end of the cylinder 31 and provided in the axial direction of the cavity (36, 38, 39) of the cylinder 31; a thick shaft portion 35 provided coaxially with the rod 32b, formed in a columnar shape having an outer diameter larger than the outer diameter of the rod 32b, and provided so as to be slidable in the large diameter region 38; a thin shaft portion 34 protruding from a distal end surface of the thick shaft portion 35 or the rod 32b in the axial direction and provided so as to be slidable in the small diameter region 36; and a guide portion 40 connected to the thin shaft portion 34 so as to protrude from a distal end of the thin shaft portion 34 in the axial direction, provided so as to be slidable in the small diameter region 36, and having a ventilation hole 41 and a lateral hole 42 formed therein as a ventilation passage.

(27) The ventilation hole 41 extends in the guide portion 40 in the axial direction, has an opening 41a having a lower end opened outward at a distal end of the guide portion 40, and has an upper end closed with a distal end edge of a distal end 34a of the thin shaft portion. The lateral hole 42 has an opening communicating with the ventilation hole 41 at the upper end of the ventilation hole 41 and formed in a side surface of the guide portion 40. Here, the sum of a length d3 of the thin shaft portion 34 and a length d4 of the ventilation hole 41 as the ventilation passage in the axial direction is longer than the stroke D of the thick shaft portion 35.

(28) That is, d3+d4>D is satisfied. Meanwhile, D>d3 is satisfied.

(29) Furthermore, in the dispensing cylinder 30 according to the present embodiment, an O-ring 37a as an airtight seal member is provided so as to surround an axis in a peripheral direction on an inner peripheral surface of an upper end portion of the small diameter region 36, and an O-ring 37b as an airtight seal member is provided so as to surround the axis in a peripheral direction on an outer peripheral surface of the thick shaft portion 35 in the thick shaft portion 35.

(30) Furthermore, in the dispensing cylinder 30 according to the present embodiment, between the large diameter region 38 and the small diameter region 36, a floating region 39 in which the thick shaft portion 35 is floatable in the axial direction is provided coaxially with the large diameter region 38 and the small diameter region 36. The floating region 39 has a length d0 in the axial direction, longer than the length d3 of the thin shaft portion 34 in the axial direction. Alternatively, in a case where the length d1 of the thick shaft portion 35 cannot be ignored, the floating region 39 has a length equal to or longer than d3+d1. In the present embodiment, the floating region 39 has an inner peripheral surface on which the thick shaft portion 35 does not slide, and has an inner diameter with a maximum diameter larger than the large diameter. Therefore, on a boundary with the large diameter region 38, a reversely tapered inclined surface is formed toward the nozzle 33. On a boundary with the small diameter region 36, at least one step projecting inward is formed toward a distal end of the nozzle so as to partition an inner wall surface of the cavity.

(31) FIG. 4 illustrates operation of the dispensing cylinder 30 according to the present embodiment. FIGS. 4(1) and 4(2) illustrate suction operation of a minute amount of liquid. In FIG. 4(1), the thick shaft portion 35 is located at a bottom dead center of a predetermined stroke of the plunger 32 as a lowermost end of the floating region 39, that is, an upper end of the small diameter region 36. Therefore, the thin shaft portion 34 and the guide portion 40 are inserted into the small diameter region 36. In this state, a distal end of a dispensing tip attached to the nozzle 33 is inserted into a container-storing liquid.

(32) Incidentally, if the stroke of the thick shaft portion 35 is represented by D, a distance from a lowermost end of the thick shaft portion 35 in the axial direction is represented by d, the length of the thin shaft portion 34 is represented by d3, thus the length d0 of the floating region 19 is equal to or longer than r0+d3 or r0+d3+d1 (in a case where the length of d1 cannot be ignored), the sum of the lengths of the small diameter region and the through hole of the nozzle is represented by d2, and the length of the guide portion 40 is represented by d4, as described above, 0<d<D, d3+d4+r0<d2, d3<D, and d3+d4+r0>D are satisfied (provided that r0=0 is satisfied in the case of FIG. 4).

(33) In FIG. 4(2), when the plunger 32 is raised from the bottom dead center by the distance d (<d3), the thin shaft portion 34 and the guide portion 40 slide in the small diameter region 36 and rise by the distance d, the thick shaft portion 35 moves by d in the floating region 39, and liquid flows into the dispensing tip attached to the nozzle 33 in an amount of S2×d.

(34) d<d3 in FIGS. 4(1) and 4(2) corresponds to the minute amount suction and discharge section.

(35) In FIG. 4(3), when the plunger 32 is moved by the distance d=d3 (<D) from the bottom dead center, the thin shaft portion 34 starts to be withdrawn from the small diameter region 36, the thick shaft portion 35 starts to be inserted into the large diameter region 38. However, the guide portion 40 is connected so as to protrude from a distal end of the thin shaft portion 34, and sliding continues while the guide portion 40 stays in the small diameter region 36. Therefore, the O-ring 37a as the airtight seal member provided in the small diameter region 36 maintains airtightness between a surface of the guide portion 40 and the O-ring 37a without being compressed by withdrawal of the thin shaft portion 34. However, communication between the small diameter region 36 and the through hole 33b of the nozzle 33 and the floating region 39 starts via the ventilation hole 41 and the lateral hole 42 as a ventilation passage provided in the guide portion 40. Suction of gas passing through the opening 33a of the nozzle 33 into the floating region 39 and the large diameter region 38 is disclosed.

(36) In FIG. 4(4), when the plunger 32 is moved from the bottom dead center by a distance d (=D>d3), the thin shaft portion 34 and the guide portion 40 move in the floating region 39, the thick shaft portion 35 slides in the large diameter region 38, and liquid is sucked into the dispensing tip attached to the nozzle 33 in an amount of S1×{d−(d0−d1)}=S1×(D−d3). Here, S1 and S2 are cross-sectional areas cut perpendicularly to the axial direction of the large diameter region 38 and the small diameter region 36, respectively. (actually, S1 and S2 also depend on the shape of a distal end of the thick shaft portion or the thin shaft portion).

(37) FIG. 5 illustrates a block diagram of a dispensing device 100 using the dispensing cylinders 10.sub.1 to 10.sub.n according to the first embodiment of the present invention.

(38) The dispensing device 100 roughly includes: a stage 20 in which a plurality of (n in this example, n≥1) container groups 20.sub.i (i=1, . . . , n) is arranged; a nozzle arrangement unit 70 in which dispensing cylinders 10.sub.i including a plurality of (n in this example) nozzles 13.sub.i for detachably attaching dispensing tips 211.sub.1 to 211.sub.n (for a minute amount) or dispensing tips 212.sub.1 to 212.sub.n (for a large amount) are arranged; a suction/discharge driving unit 53 for moving a plunger of each of the dispensing cylinders in the vertical direction and causing the dispensing tips to suck and discharge liquid; a tip detaching mechanism 59 capable of detaching the dispensing tip 211.sub.i detachably attached to the nozzle 13.sub.i; a magnetic force unit 57 capable of applying a magnetic field to the dispensing tips 211.sub.i attached to the nozzles 13 arranged in the nozzle arrangement unit 70; a dispensing head 50 having a nozzle Z-axis moving mechanism 75 capable of moving the nozzle arrangement unit 70 in the Z-axis with respect to the container group 20.sub.i; a dispensing head moving mechanism 51 capable of moving the dispensing head 50 relatively to the container group, for example, with respect to the Y-axis direction; a temperature controller 29 for performing a predetermined temperature control on a reaction container group 23.sub.i of the container group; a CPU+program+memory 60 including CPU, ROM, RAM, various external memories, a communication function such as LAN, and a program stored in ROM or the like; and an operation panel 64 including a display unit such as liquid crystal display and an operation unit such as an operation key or a touch panel.

(39) Here, the dispensing head moving mechanism 51 and the nozzle Z-axis moving mechanism correspond to the nozzle moving mechanism.

(40) Each of the container groups 20.sub.i includes: a reaction container the temperature of which can be controlled; liquid-storing units capable of storing various reagents, various specimens, a magnetic particle suspension, or the like; and tip-storing units for storing the respective dispensing tips 211.sub.i in a state of being able to be attached to attachment portions formed at distal ends of the nozzles 13.

(41) The CPU+program+memory 60 includes: an extraction control unit 61 for controlling extraction of a nucleic acid or a fragment thereof; a nucleic acid amplification control unit 62 for giving an instruction for an amplification treatment on an extracted nucleic acid; and a minute amount/large amount judgement and instruction unit 63 for judging whether a predetermined amount of liquid to be sucked and discharged is a minute amount or a large amount based on an instruction from the extraction control unit 61, the nucleic acid amplification control unit 62, or the operation panel 64 and giving an instruction to the suction/discharge driving unit 53. The extraction control unit 61 gives an instruction of extraction to the dispensing head moving mechanism 51, the nozzle Z-axis moving mechanism 75, the tip detaching mechanism 59, the temperature controller 29, the magnetic force unit 57, or the minute amount/large amount judgement and instruction unit 63. The nucleic acid amplification control unit 62 controls the dispensing head moving mechanism 51, the tip detaching mechanism 59, the suction/discharge driving unit 53, the nozzle Z-axis moving mechanism 75, and the minute amount/large amount judgement and instruction unit 63.

(42) Hereinafter, a more specific embodiment example of the above-described dispensing device 100 according to the first embodiment of the present invention will be described with reference to FIGS. 6 to 8.

(43) FIG. 6 is an overall perspective view of the dispensing device 100 according to an embodiment example of the present invention.

(44) For example, the dispensing device 100 has a size of about 600 mm in a longitudinal direction (Y-axis direction), a transverse direction (X-axis direction), and a height (Z-axis direction). On the stage 20, mainly, the container groups 20.sub.1 to 20.sub.8 (n=8), the dispensing head 50 capable of moving in a serial arrangement direction (Y-axis direction) with respect to the container groups 20.sub.1 to 20.sub.8, the dispensing head moving mechanism 51 for moving the dispensing head 50 in the Y-axis direction, and the temperature controller 29 are provided on the stage 20. Note that the operation panel 64 and the CPU+program+memory 60 are attached to a housing (not illustrated) for housing these container groups 20.sub.1 to 20.sub.8 and the dispensing head 50.

(45) The dispensing head 50 includes the nozzle arrangement unit 70 in which eight dispensing cylinders 10, provided so as to be movable in the vertical direction (Z-axis direction) with respect to a Y-axis moving frame 512 provided so as to be movable in the serial arrangement direction (Y-axis direction) are arranged in the X-axis direction at a predetermined pitch (for example, 18 mm) and the nozzles 13 (to which eight dispensing tips 211.sub.i can be attached) provided in the dispensing cylinder 10.sub.i (i=1 to 8, omitted hereinafter).

(46) The dispensing head moving mechanism 51 includes: a Y-axis moving motor 511; a coupling tool 513 connected to a timing belt 515 driven by the Y-axis moving motor 511 to be movable in the Y-axis direction by movement of the timing belt 515; and two rotatable pulleys 514 engaged with the timing belt 515.

(47) The nozzle arrangement unit 70 of the dispensing head 50 includes support plates 701 and 702 for supporting the cylinders 10.sub.1 to 10.sub.8 and the nozzles 13 communicating with the cylinders so as to arrange the cylinders 10.sub.1 to 10.sub.8 and the nozzles 13 at the above pitch, and the support plates 701 and 702 are supported so as to be movable in the Z-axis direction with respect to the Y axis moving frame 512. A Z-axis moving motor for moving the nozzle arrangement unit 70 in the Z-axis direction is provided in the Y-axis moving frame 512.

(48) A plunger driving plate 532 for vertically driving the eight plungers 12 slidable in the dispensing cylinders 10.sub.1 to 10.sub.8 arranged in the nozzle arrangement unit 70 and a suction and discharge driving motor 531 for driving the plunger driving plate 532 are provided. Here, the plunger driving plate 532 and the motor 531 correspond to the suction/discharge driving unit 53.

(49) A tip detaching member 591 is provided below the nozzle arrangement unit 70. The tip detaching member 591 is attached to lower ends of two shafts 592 supported by the nozzle arrangement unit 70 and movable downward while being urged upward, at both end portions thereof, and is thereby supported horizontally by the nozzle arrangement unit 70. An upper end 593 of each of the shafts 592 is located above the upper ends of the dispensing cylinders 10.sub.1 to 10.sub.8 but below the bottom dead center of the stroke (D) of normal suction and discharge of the plunger driving plate 532. The length of each of the shafts 592 is related to the above-described “r0 (≠0)”. The tip detaching mechanism 59 is provided in which the plunger driving plate 532 is lowered to the vicinity of upper ends of the dispensing cylinders 10.sub.1 to 10.sub.8 beyond the stroke (D), the upper ends 593 of the shafts 592 are thereby pushed downward, the shafts 592 are lowered to the vicinity of upper ends of the dispensing cylinders 10.sub.1 to 10.sub.8, and the tip detaching member 591 is thereby pushed downward and lowered. In the tip detaching member 591, eight holes each having an inner diameter larger than the outer diameter of an attachment portion of the nozzle 13 but smaller than the maximum outer diameter of the dispensing tip 211.sub.i, are formed at the above pitch so that the nozzles 13 penetrate through the eight holes.

(50) The magnetic force unit 57 is obtained by providing ten magnets 571 capable of applying a magnetic field to an inside of the dispensing tip 211.sub.i and removing the magnetic field therefrom by being provided so as to be able to come into and out of contact with a small diameter portion 211.sub.ia of the dispensing tip 211.sub.i in a movable body 572 movable in the Y-axis direction. Note that one magnet on each side is provided in order to apply a uniform magnetic field to each of the dispensing tips 211, and is different from eight magnets provided so as to be able to come into and out of each of the dispensing tips.

(51) As illustrated in FIG. 9, in each of the container groups 20.sub.1 to 20.sub.8 on the stage 20, for example, a cartridge container 24.sub.i in which 14 reaction containers or various storing units are arranged in series, a cartridge container 28.sub.i in which four various storing units are arranged in series, and a parent specimen tube 26.sub.i are arranged. The container groups are arranged in parallel with each other in the serial arrangement direction such that the same type of storing units, the same type of reaction containers, and the same type of parent specimen tubes are located at the same positions respectively in the serial arrangement direction (Y-axis direction).

(52) Here, the cartridge container 24.sub.i includes two reaction containers 23.sub.i having different volumes, a liquid-storing unit group 27i of ten pre-packed or emμLy liquid-storing units, and a tip-storing unit group 210.sub.i for storing two dispensing tips 211.sub.i and 212.sub.i. The cartridge container 28.sub.i includes a storing unit for storing two scattering prevention plugs 221.sub.ia and a storing unit for storing piercing tips 213.sub.i and 214.sub.i. Note that the scattering prevention plugs 221.sub.i are used for sealing a container (not illustrated) to which ultrasonic waves can be applied, applying ultrasonic waves or the like to a sample or the like inside, and crushing a cell to facilitate extraction of a nucleic acid, and attached to the nozzle 13 so as to be movable.

(53) Subsequently, regarding operation of the dispensing device 100 according to the present example, a series of treatment operations until real-time PCR of a nucleic acid of a specimen containing bacteria is performed will be described below.

(54) In step S1, a specimen to be tested, various cleaning liquids, and various reagents are supplied onto the stage 20 in advance, and liquid-storing units pre-packed with reagents or the like is attached thereto.

(55) By operating a touch panel or the like of the operation panel 64, an instruction to start a separation and extraction treatment and an amplification treatment is made.

(56) Then, in step S2, the extraction control unit 61 provided in the CPU+program+memory 60 of the dispensing device 100 instructs the dispensing head moving mechanism 51 to move the dispensing head 50 in the Y-axis direction to locate the dispensing head 50 in a corresponding tip-storing unit of a cartridge container 28.sub.i of each of the container groups 20.sub.i. The piercing tip 213 is attached to the nozzle 13 by the nozzle Z-axis moving mechanism 75. The dispensing head 50 is further moved in the Y-axis direction to locate the piercing tip 213.sub.i above the first liquid-storing unit of the liquid-storing unit group 27.sub.i of the container group, and the nozzle is lowered by the nozzle Z-axis moving mechanism 75 to pierce a film covering an opening of the liquid-storing unit. Similarly, the dispensing head 50 is moved in the Y-axis direction to sequentially pierce the other liquid-storing units of the liquid-storing unit group 27.sub.i and the reaction container group 23.sub.i, and the piercing tip 213.sub.i is detached into the tip-storing unit by the tip detaching mechanism 59.

(57) In step S4, the dispensing head 50 is again moved in the Y-axis direction and moved to a tip or the like-storing unit group 21.sub.i. The respective nozzles 13.sub.i are lowered by the nozzle Z-axis moving mechanism 75, and the dispensing tip 211.sub.i is attached thereto. Next, the dispensing tip 211.sub.i is raised by the nozzle Z-axis moving mechanism 75 and then moved along the Y-axis by the dispensing head moving mechanism 51 to reach the eighth liquid-storing unit of the liquid-storing unit group 27.sub.i, and a predetermined amount of isopropanol is sucked from the liquid-storing unit. The dispensing tip 211.sub.i is moved again along the X-axis, and dispensing is performed into a solution component (NaCl and SDS solution) stored in each of the third and fifth liquid-storing units and distilled water stored in the sixth liquid-storing unit in a predetermined amount. As a result, 500 μL of a bonding buffer solution (NaCl, SDS, and isopropanol), 700 μL of cleaning liquid 1 (NaCl, SDS, and isopropanol), and 700 μL of cleaning liquid 2 (water: 50%, isopropanol: 50%) are prepared in the third, fifth, and sixth liquid-storing units as separation and extraction solutions, respectively.

(58) At this time, the minute amount/large amount judgement and instruction unit 63 judges that the predetermined amount is a large amount, and the thick shaft portion 15 is located in the large amount suction and discharge section to slide in the large diameter region 18 by a distance D corresponding to the predetermined amount.

(59) In step S5, the dispensing tip 211.sub.i is moved to the parent specimen tube 26.sub.i storing a specimen separately, in the tip or the like-storing unit group 21.sub.i. Thereafter, the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i is lowered and inserted thereinto using the nozzle Z-axis moving mechanism 75, and the driving plate 532 of the suction/discharge driving unit 53 is raised and lowered. Suction and discharge of a suspension of the specimen stored in the parent specimen tube 26.sub.i are thereby repeated to make the specimen suspended in the liquid. Thereafter, the specimen suspension is sucked into the dispensing tip 211.sub.i. The specimen suspension is moved to the first liquid-storing unit of the liquid-storing unit group 27.sub.i storing Lysis 1 (enzyme) as a separation and extraction solution along the Y-axis by the dispensing head moving mechanism 51. The small diameter portion 211.sub.ia of the dispensing tip 211.sub.i is inserted thereinto through a hole of a pierced film, and suction and discharge are repeated in order to stir the specimen suspension and the Lysis 1.

(60) In step S6, the whole amount of the stirred liquid is sucked by the dispensing tip 211.sub.i to be stored in the reaction container 232.sub.i formed of a reaction tube held in the storing hole set at 55° C. by the constant temperature controller, and is incubated. As a result, a protein contained in the specimen is destroyed to reduce the molecular weight thereof. After a lapse of a predetermined time, the dispensing tip 211.sub.i is moved to the second liquid-storing unit of the liquid-storing unit group 27.sub.i by the dispensing head moving mechanism 51 while the reaction solution is left in the reaction tube. The whole amount of the liquid stored in the second liquid-storing unit is sucked using the nozzle Z-axis moving mechanism 75 and the suction/discharge driving unit 53, and transferred by the dispensing head moving mechanism 51 using the dispensing tip 211.sub.i. The small diameter portion is inserted into the third liquid-storing unit through the hole of the film to discharge the reaction solution.

(61) In step S7, a bonding buffer solution as a separation and extraction solution stored in the third liquid-storing unit and the reaction solution are stirred to further dehydrate a solubilized protein, and a nucleic acid or a fragment thereof is dispersed in the solution.

(62) In step S8, the small diameter portion of the dispensing tip 211.sub.i is inserted into the third liquid-storing unit through the hole of the film using the dispensing tip 211.sub.i, and the whole amount is sucked. The dispensing tip 211.sub.i is raised by the nozzle Z-axis moving mechanism 75. The reaction solution is transferred to the fourth liquid-storing unit. A magnetic particle suspension stored in the fourth liquid-storing unit and the reaction solution are stirred. A cation structure in which a Na.sup.+ ion is bonded to a hydroxy group formed on surfaces of magnetic particles contained in the magnetic particle suspension is formed. Therefore, negatively charged DNA is caμLured by the magnetic particles.

(63) In step S9, by making the magnet 571 of the magnetic force unit 57 approach the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i, the magnetic particles are adsorbed by an inner wall of the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i. The dispensing tip 211.sub.i is raised by the nozzle Z-axis moving mechanism 75 while the magnetic particles are adsorbed by the inner wall of the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i. The dispensing tip 211.sub.i is moved from the fourth liquid-storing unit to the fifth liquid-storing unit using the dispensing head moving mechanism 51, and the small diameter portion 211.sub.ia is inserted thereinto through the hole of the film.

(64) While a magnetic force to an inside of the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i is removed by separating the magnet 571 of the magnetic force unit 57 from the small diameter portion 211.sub.ia, a cleaning liquid 1 (NaCl, SDS, and isopropanol) stored in the fifth liquid-storing unit is repeatedly sucked and discharged. The magnetic particles are thereby detached from the inner wall, are stirred in the cleaning liquid 1, and a protein is thereby cleaned. Thereafter, while the magnetic particles are adsorbed by the inner wall of the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i by making the magnet 571 of the magnetic force unit 57 again approach the small diameter portion 211.sub.ia, the dispensing tip 211.sub.i is moved by the nozzle Z-axis moving mechanism 75 from the fifth liquid-storing unit to the sixth liquid-storing unit by the dispensing head moving mechanism 51.

(65) In step S10, the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i is inserted through the hole of the film using the nozzle Z-axis moving mechanism 75. While the magnetic force to the inside of the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i is removed by separating the magnet 571 of the magnetic force unit 57 from the small diameter portion 211.sub.ia, a cleaning liquid 2 (isopropanol) stored in the sixth liquid-storing unit is repeatedly sucked and discharged. The magnetic particles are thereby stirred in the liquid, NaCl and SDS are removed, and a protein is cleaned. Thereafter, while the magnetic particles are adsorbed by the inner wall of the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i by making the magnet 571 of the magnetic force unit 57 again approach the small diameter portion 211.sub.ia, the dispensing tip 211.sub.i is raised by the nozzle Z-axis moving mechanism 75, and then moved from the sixth liquid-storing unit to the seventh liquid-storing unit storing distilled water by the dispensing head moving mechanism 51.

(66) In step S11, the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i is lowered by the nozzle Z-axis moving mechanism 75 through the hole. By repeating suction and discharge of the distilled water at a slow flow rate while the magnetic force is applied to the inside of the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i, the cleaning liquid 2 (isopropanol) is replaced with the water and removed. Thereafter, by sucking and discharging the magnetic particles repeatedly in distilled water as the dissociated solution while the magnet 571 of the magnetic force unit 57 is separated from the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i and the magnetic force is removed, stirring is performed, and a nucleic acid retained by the magnetic particles or a fragment thereof is dissociated (eluted) from the magnetic particles into the liquid. Thereafter, by making the magnet 571 approach the small diameter portion 211.sub.ia of the dispensing tip 211.sub.i, the magnetic field is applied to the inside of the small diameter portion, the magnetic particles are adsorbed by the inner wall, and the solution containing the extracted nucleic acid or the like is left in the eighth liquid-storing unit. The dispensing tip 211.sub.i is moved to the storing unit in which the dispensing tip 211.sub.i of the tip or the like-storing unit group 21.sub.i was stored by the dispensing head moving mechanism 51. The dispensing tip 211.sub.i which has adsorbed the magnetic particles is detached from the nozzle 71.sub.i into the storing unit together with the magnetic particles using the detaching member 591 of the tip detaching mechanism 59.

(67) Subsequently, steps S12 to S15 correspond to a nucleic acid amplification step.

(68) In step S12, a new minute amount dispensing tip 212.sub.i is attached to the nozzle 13.sub.i. The solution containing the nucleic acid or the like stored in the eighth liquid-storing unit is sucked, transferred to the PCR tube 231.sub.i storing an amplification solution 234.sub.i in advance, discharged, and introduced into the container. Hereinafter, in accordance with a predetermined procedure, in the PCR tube 231.sub.i, the nucleic acid amplification control unit 62 instructs the temperature controller 29 to repeat a temperature control cycle by real time PCR, for example, a cycle of heating the PCR tube 231.sub.i at 96° C. for five seconds and heating the PCR tube 231.sub.i at 60° C. for 15 seconds, for example, 49 times.

(69) Next, in order to compare the accuracy of the dispensing cylinder according to the present embodiment with a conventional cylinder, an operation of sucking and discharging a predetermined amount (for example, 5 μL and 100 μL) of liquid was performed using the dispensing device 100. A coefficient of variation indicating how much the amount of liquid actually sucked and discharged varies from a target predetermined amount was measured.

(70) Results thereof are illustrated below.

(71) TABLE-US-00001 5 μL dispensing with 2.5 mm φ dispensing cylinder according to the present embodiment Acquired data (μL) Lane1 Lane2 Lane3 Lane4 Lane5 Lane6 Lane7 Lane8 N = 1 5.1 5.0 4.9 4.8 4.9 4.7 4.8 5.1 N = 2 5.0 5.1 4.9 4.8 4.9 4.8 4.9 4.8 N = 3 4.8 4.8 4.8 4.7 4.9 4.8 4.7 4.7 Average Standard deviation Coefficient of variation (CV %) 4.8 0.12 2.5%

(72) TABLE-US-00002 100 μL dispensing with 10 mm φ dispensing cylinder according to the present embodiment Acquired data (μL) Lane1 Lane2 Lane3 Lane4 Lane5 Lane6 Lane7 Lane8 N = 1 99.8 99.3 99.3 99.5 100.0 100.2 100.4 101.1 N = 2 99.2 98.8 99.3 99.5 99.2 99.8 99.7 99.6 N = 3 100.0 99.1 99.5 99.5 97.7 99.7 100.0 100.6 Average Standard deviation Coefficient of variation (CV %) 99.6 0.64 0.65%

(73) Meanwhile, a measurement example is illustrated in a case where 5 μL as a predetermined amount is sucked and discharged using a device including eight rows of container groups having conventional cylinders incorporated therein.

(74) TABLE-US-00003 5 μL dispensing with 4 mm φ dispensing cylinder according to conventional example Acquired data (μL) Lane1 Lane2 Lane3 Lane4 Lane5 Lane6 Lane7 Lane8 N = 1 5.7 5.3 5.3 5.9 5.0 5.0 5.7 5.6 N = 2 5.5 5.3 5.8 6.0 5.4 5.3 5.7 5.7 N = 3 5.3 5.4 5.1 6.0 5.5 5.7 5.6 5.9 Average Standard deviation Coefficient of variation (CV %) 5.5 0.29 5.2%

(75) Comparison among the above coefficients of variation clearly indicates that the dispensing cylinder according to the present embodiment has a small coefficient of variation and high accuracy even when a minute amount of liquid is dispensed.

(76) A reason for this is as follows. That is, in a case of dispensing a minute amount of liquid and in a case of dispensing a large amount of liquid, in a dispensing cylinder according to a conventional example, a single dispensing cylinder is used, and the dispensing cylinder has a plunger and a cavity having the same cross sectional area. Meanwhile, in the dispensing cylinder according to the present embodiment, although a single dispensing cylinder is used, in a case of dispensing a minute amount of liquid, a thin shaft portion having a sufficiently smaller cross sectional area than the cross sectional area is slid, and in a case of dispensing a large amount of liquid, a thick shaft portion having a sufficiently larger sectional area than the cross sectional area is slid. Therefore, regarding a minute amount of liquid, a sufficient sliding distance can be ensured, and therefore highly accurate dispensing is possible. Furthermore, in a case of dispensing a large amount of liquid, a sliding distance can be reduced, and therefore a compact dispensing cylinder can be provided. Actually, in the dispensing cylinder according to the present embodiment, even in a case of a minute amount (5 μL) or a large amount (100 μL), the sliding distance is about 1 mm. Therefore, highly accurate measurement is possible. In order to dispense 1000 μL of liquid, a sliding distance of at most 13 mm is enough. Meanwhile, in a device according to a conventional example, in a case of a small volume, the sliding distance is only about 0.5 mm, but in a case of a large volume (100 μL), the sliding distance is about 9 mm, and the sliding distance is about 90 mm in order to dispense 1000 μL of liquid. This means that the scale of the device may be expanded as compared with the present embodiment.

(77) The above-described embodiments have been described specifically for the purpose of better understanding of the present invention, and do not limit another embodiment. Therefore, the above-described embodiments can be modified within a range not changing the gist of the invention. For example, as an embodiment, only the case where the airtight seal members are provided in the small diameter region and the thick shaft portion has been described. However, the present invention is not limited to this case. Needless to say, the present invention can also be applied to a case where the airtight seal members are provided in the thin shaft portion and the thick shaft portion, a case where the airtight seal members are provided in the thin shaft portion and the large diameter region, and a case where the airtight seal members are provided in the small diameter region and the large diameter region. In the above description, the case of r0=0 is illustrated in the drawings, but it is sufficiently possible to estimate a case of r0≠0 from the drawings. Only the dispensing cylinder 10.sub.i according to the first embodiment has been described as an example of the dispensing device. However, needless to say, the dispensing cylinder 30.sub.i according to the second embodiment can be used. As an example of the ventilation passage, the case where only the ventilation hole and the lateral hole are present has been described. However, the present invention can also be applied to a case where a groove and a slit are formed. Regarding the dispensing device, an example of a nucleic acid treatment has been described, but the present invention is not limited to this example. Needless to say, the present invention can also be applied to a treatment of a protein or the like.

(78) In addition, the numerical values, the number of times, the shape, the number (for example, the number of the dispensing cylinders used in the dispensing device is not limited to eight), the amount, and the like are not limited to those in these cases.

INDUSTRIAL APPLICABILITY

(79) The present invention relates to a dispensing cylinder, and a dispensing device and a dispensing treatment method using the dispensing cylinder, performs dispensing of a specimen collected from a patient or the like, a treatment thereof, a test thereof, optical measurement thereof, and recording thereof, and can be used particularly in a field requiring handling of a biopolymer such as a gene, an immune system, an amino acid, a protein, or a sugar, and a low molecular biological substance, for example, in various fields such a biochemistry field, an industrial field, an agriculture field such as food, agriculture, or fishery processing, a pharmaceutical field, and a medical field such as hygiene, health, immunity, diseases, or genetics.

REFERENCE SIGNS LIST

(80) 10, 30 Dispensing cylinder 10.sub.1 to 10.sub.n Dispensing cylinder 11, 31 Cylinder (main body) 12, 32 Plunger 13, 33 Nozzle 14, 34 Thin shaft portion 15, 35 Thick shaft portion 16, 36 Small diameter region 17, 37 Airtight seal member 18, 38 Large diameter region 19, 39 Floating region 20 Stage 20.sub.1 to 20.sub.n Container group 211i to 211.sub.n Large amount dispensing tip 212.sub.1 to 212.sub.n Minute amount dispensing tip 40 Guide portion 41 Ventilation hole (ventilation passage) 42 Lateral hole (ventilation passage) 50 Dispensing head 51 Dispensing head moving mechanism (nozzle moving mechanism) 59 Tip detaching mechanism 60 CPU+program+memory 100 Dispensing device