Patent Application
20200240921
The present invention relates to and asserts priority from Japanese patent application No. 2019-011323 filed on Jan. 25, 2019, and incorporates the entirely of the contents and subject matter of all the above application herein by reference.
The present invention relates to a technology on an apparatus, a system, and a method for luminescence measurement.
Mixing a specimen and a luminescent reagent in a microplate causes chemiluminescence and, evaluating an amount of the luminescence allows to obtain biological information of the specimen. To detect such a luminescence at higher sensitivity, a Photomultiplier Tube (PMT) is generally often used as a luminescence detector. Further, in most cases, dispensing a luminescent reagent and measuring luminescence are performed in the same apparatus, and both a dispensing mechanism and a measuring mechanism are mounted together in the same apparatus.
Vertical positioning of a nozzle that is used to dispense a luminescent reagent is performed according to a specimen, a type of a luminescent reagent, its liquid surface level, and a shape of a microplate. Further, the photosensitive surface of a photomultiplier tube can receive light at higher sensitivity as positioned closer to a luminous point, and therefore, the photomultiplier tube is generally positioned as close to the microplate as possible to perform luminescence measurement.
A chemiluminescence measurement apparatus using a luminescent reagent called “flash type” requires performing luminescence measurement as early as possible (typically, within several seconds) after mixing a luminescent reagent and a specimen. To do so, it is desirable to arrange a nozzle for dispensing the luminescent reagent and a photomultiplier tube for measuring luminescence as close to each other as possible. In most of chemiluminescence measurement apparatuses that are commonly used today the nozzle and the photomultiplier tube are placed on the same stage, with the aims of high throughput, downsizing the apparatus, and cost reduction, as illustrated in Nonpatent Literature 1 (refer to
Moreover, Patent Literature 1 discloses a dispensing apparatus and an analysis apparatus equipped therewith described as: “a dispensing apparatus of the present invention comprises a dispensing tip 2, a syringe part 3, and a syringe base 4. The syringe part 3 includes a nozzle 11 extending in a vertical direction and having the dispensing tip 2 attached to its bottom end in the vertical direction. The syringe base 4 is provided with an elastic member 8 which supports the syringe part 3 movably and urges the syringe part 3 upward in the vertical direction. The syringe part 3 and the syringe base 4 are movable with respect to each other in the vertical direction.”
Nonpatent Literature 1: “Navigator System OPERATING MANUAL Instructions for Use of Products GM2000 and GM2010” (Online), GloMax, (Searched out on Dec. 28, 2018) Internet <URL: https://www.promega.jp/-/media/files/resources/protocols/technical-manuals/101/glomax-navigator-system-operating-manual.pdf?la=en>
As described in Nonpatent Literature 1, when the nozzle and the photomultiplier tube are fixed on the same stage, “adjusting a height of the nozzle” and “adjusting a height of the photomultiplier tube” interlock with each other and the height of each part cannot be adjusted independently. As a result, dispensing and measurement cannot be performed except at a height designed in advance and a specification change of a microplate makes a design change needed.
Accordingly, a technology described in Nonpatent Literature 1 poses the following problems:
(A1) Redesign may be needed when the specifications of a microplate (such as depth) are changed;
(A2) Even if a contact of the nozzle with a specimen should not be allowed, a change in a liquid quantity (liquid level) of a specimen may cause the contact
(A3) When a tip of the nozzle is positioned higher than a top surface of a plate, a luminescent reagent may spatter on the surface of the plate, when being dispensed.
(A4) When the photomultiplier tube floats from a well of a microplate, its sensitivity degrades and, besides, incident light from an adjacent well may induce luminous crosstalk.
(A5) When a measurement is performed for a well positioned at an end of a microplate under a condition of the tip of the nozzle being positioned lower than a photosensitive surface of the photomultiplier tube, the nozzle may hit against, for example, the top surface (deck) of an end portion of the microplate or the like when the photomultiplier tube moves down, which may contaminate the nozzle.
The present invention is developed in view of the technical background and the above-described problems, and it is an object of the invention to provide an apparatus performing an efficient luminescence measurement.
To solve the above-noted problem, the present invention provides an apparatus comprising: a nozzle for dispensing liquid into a well in a microplate, a luminescence measurement unit configured for measuring luminescence occurring in the well, and a moving unit configured for moving the nozzle and the luminescence measurement unit together vertically and horizontally, wherein the nozzle is secured to the moving unit, and the luminescence measurement unit is mounted to be vertically movable with respect to the moving unit.
Other solutions are described, as appropriately, in Description of Embodiments below.
The present invention enables a more efficient luminescence measurement.
Next, embodiments for carrying out the present invention (referred to as simply “embodiments”) are described in detail with reference to the drawings as appropriately as needed. Note that exaggeration, deformation, etc. are made in some of the figures and dimensions of each part in the figures are not always consistent with each other.
As depicted in
As depicted in
In addition, as depicted in
Thereby, the luminescence measurement unit 120 is allowed to move vertically with respect to the holder 111 by being guided by the supporting posts 11 and the holes serving. Furthermore, each supporting post 113 is inserted through a spring 114. One end of the spring 114 is fixed to the luminescence measurement unit 120 and the other end is fixed to the top board 116 of the holder 111.
Additionally, the luminescence measurement unit 20 is provided at the bottom surface thereof with a photosensitive portion 122 to receive light emitted by a mixture liquid 302, as depicted in
Further, pawls 115 are provided in the opening in the bottom surface of the holder 111. Also, first protrusions 121 are provided in lateral surfaces of the luminescence measurement unit 120. The first protrusions 121 of the luminescence measurement unit 120 are lathed by the pawls 115 so that the luminescence measurement unit 120 are prevented from falling from the holder 111 or can be avoided from extending from the holder 111 more than necessary.
The stage 101 is fixedly provided with a nozzle position adjusting part 130. The nozzle position adjusting part 130 is described below. The nozzle position adjusting part 130 is provide with a nozzle 140 to dispense a luminescent reagent (liquid) or the like into a well W of the microplate M. That is, the stage 101 is provided with the nozzle 140 via the nozzle position adjusting part 130. Because the nozzle position adjusting part 130 is fixed to the stage 101, the nozzle 140 moves accompanying with movement of the stage 101. Note that the nozzle 140 is essential, but the nozzle position adjusting part 130 may be dispensed with.
The nozzle 140 is connected to a syringe which is not depicted. The syringe is controlled to regulate the discharge of the luminescent reagent through the nozzle 140.
Also, as depicted in
The chemiluminescence measurement system Z includes the chemiluminescence measurement apparatus 1 described with
The control device (control unit) 2 may be a PC (Personal Computer), a PLC (programmable Logic Controller) or the like, and includes a memory 201, a CPU 202, and a communication device 203 for communicating with the chemiluminescence measurement apparatus 1.
The memory 201 includes a program stored in and loaded from a storage device which is not depicted. The loaded programs are then executed by the CPU 202 and, thereby, a processing unit 201 and a movement controller 211, a dispensing controller 212, and a measurement controller 213 which are included in the processing unit 201 are embodied.
The movement controller 211 controls movement of the stage 101.
The dispensing controller 212 controls the syringe (not depicted) to dispense a luminescent reagent or the like.
The measurement controller 213 controls luminescence measurement performed by the luminescence measurement unit 120.
Next, with reference to
Here is assumed that the chemiluminescence measurement apparatus 1 is in a state that a specimen 301 has been dispensed in wells W of the microplate M and a mixture liquid 302 is produced by dispensing a luminescent reagent from the nozzle 140 into the specimen 301. The specimen 301 may be dispensed by either human or through the nozzle 140. If the specimen 301 is dispensed by the nozzle 140, the nozzle 140 is replaced after dispensing the specimen 301. Here, when a luminescent reagent is dispensed into the specimen 301, chemical reaction of the specimen 301 with the luminescent reagent makes the mixture liquid 302 produce luminescence. Measuring an amount of such luminescence enables the luminescence measurement unit 120 to measure a quantity of an intended substance.
In
First, as depicted in
Next, the movement controller 211 moves the stage 101 down, as indicated by a blank arrow in
Then, the luminescence measurement unit 120 contacts a deck D of the microplate M, as depicted in
This results in shrinking of the springs 114 through which the supporting posts 113 are inserted, as depicted in
Then, the dispensing controller 212 makes the nozzle 140 to dispense the luminescent reagent into the well Wb. This results in a mixture liquid 302 in the well Wb. Also, at the same time of dispensing the luminescent reagent by the nozzle 140, luminescence produced by the mixture liquid 302 in the well Wa is measured by the luminescence measurement unit 120.
When the dispensing and the measurement are finished, the movement controller 211 moves the stage 101 upward, as indicated by a blank arrow in
When the nozzle 140 and the luminescence measurement unit 120 move up to the previous position before they are set over the respective wells W, the movement controller 211 horizontally moves the stage 101, as indicated by a blank arrow in
That is, the movement controller 211 horizontally moves the stage 101 to position the luminescence measurement unit 120 just above the well Wb into which the luminescent reagent is dispensed by the nozzle 140 and the nozzle 140 just above the well We into which the luminescent reagent is dispensed next.
Further, the microplate M is horizontally moved in a direction indicated by a bold arrow in
Subsequently, by repeating the operation illustrated in
Note that an interval between the nozzle 140 and the luminescence measurement unit 120 is adjusted beforehand by the nozzle position adjusting part 130 so as to be equal to an interval between wells W.
First, a user sets the microplate M with the specimen 301 dispensed (S101). Further, the user pours a luminescent reagent into a cylinder (not depicted) which is connected to the nozzle 140 (S102). Furthermore, the user performs a horizontal position adjustment of the nozzle 140 using the nozzle position adjusting part 130, if necessary.
Next, the movement controller 211 horizontally moves the stage 101 to position the nozzle 140 just above the well W into which the luminescent reagent is first dispensed (S111).
Then, the movement controller 211 moves the stage 101 down to a position appropriate for dispensing (S112). The nozzle position sensor 160 detects whether or not the position is appropriate. It should be noted that an appropriate position differs depending on measurement conditions. Thus, detecting whether or not the position is appropriate by the nozzle position sensor 160 allows adjustment by the user to be unnecessary and to improve throughput.
Subsequently, the dispensing controller 212 causes the nozzle 140 to dispense the luminescent reagent (S113). Note that because there is no well W for which luminescence measurement can be performed at the stage of step S113, luminescence measurement is not performed.
When the dispensing finishes, the movement controller 211 horizontally moves the stage 101 (S121). Here, after moving the stage 101 up, the movement controller 211 horizontally moves the stage 101 to position the nozzle 140 just above the well W into which the luminescent reagent is to be dispensed next and position the luminescence measurement unit 120 just above the well W into which the luminescent reagent is dispensed.
Next, the movement controller 211 moves the stage 101 down (S122;
When the dispensing and luminescence measurement finish, the movement controller 211 moves the stage 101 up until the nozzle 140 and the luminescence measurement unit 120 arrive at the position before they are set over the respective wells W (
After that, the dispensing controller 212 determines whether dispensing the reagent into all wells is complete or not (S131).
As a result of step S131, if it is determined that dispensing the reagent into all the wells is not complete (S131→No), the processing unit 210 causes the process to return to the step S121.
As a result of the step S131, if it is determined that dispensing the reagent into all the wells is complete (S131→Yes), the movement controller 211 horizontally moves the stage 101 to position the luminescence measurement unit 120 just above the last well W into which the luminescent reagent is dispensed (S132).
Then, the movement controller 211 moves the stage 101 down (S133).
When moving down the stage 101 finishes, the measurement controller 213 causes the luminescence measurement unit 120 to perform luminescence measurement (S134). Here, the finish of moving down of the stage 101 is determined by the position of the nozzle 140, which position is detected by the nozzle position sensor 160. Besides, because there is no well into which the reagent is to be dispensed in the step S134, luminescence measurement only is performed.
Note that here is assumed that dispending and luminescence measurement are performed for wells forming a single line; the steps S111 through S134 may be, however, repeated for wells W in all lines formed on the microplate M.
Then, the movement controller 211 horizontally moves the stage 101 (S141). Here, the movement controller 211 horizontally moves the stage 101 so that the nozzle 140 arrives at a nozzle cleaning part which is not depicted.
After that, cleaning the nozzle 140 by the nozzle cleaning part, not depicted, is performed (S142).
Additionally, the first nozzle support 135 has its front-end face fixed with a second nozzle support 136. And, the nozzle 140 is inserted through the second nozzle support 136 so that the nozzle 140 is secured to the nozzle position adjusting part 130.
Also, the first support 131 is provided with a knob 137 at its reverse side of a side opposite to the second support 132. The knob 137 is connected to the feed screw 133.
When the user turns the knob 137, the feed screw 133 rotates. With the rotation of the feed screw 133, the first nozzle support 135 moves horizontally (in the y-axis direction). With movement of the first nozzle support 135, the second nozzle support 136 moves and, thereby, the nozzle 140 moves horizontally (in the y-axis direction). This means that the nozzle position adjusting part 130 is operated by hand.
Note that the structure of the nozzle position adjusting part 130 depicted in
Besides, the nozzle position adjusting part 130 may be a linear actuator that can move linearly in the y-axis direction. In this case, the linear actuator may be moved according to a command entered via an input device, which is not depicted, of the control device 2 to adjust the horizontal position of the nozzle 140.
As depicted in
As noted above, a pitch between wells W in the microplate M depends on specifications, manufacturers, and the like. Because the horizontal position of the nozzle 140 can be adjusted by the provision of the nozzle position adjusting part 130, it is possible to make an interval between the luminescence measurement unit 120 and the nozzle 140 keep a pitch between wells W. That is, it is possible to readily respond to a specification change of the pitch between the wells W.
As described above, the provision of the nozzle position adjusting part 130 allows to easily make the positional relation between the nozzle 140 and the luminescence measurement unit 120 equal to an interval between wells in the microplate M. Thus, it is possible to easily perform a dispensing operation into a next well W, while performing the luminescence measurement by the luminescence measurement unit 120. This results in the improvement in the throughput.
According to the first embodiment, the luminescence measurement unit 120 is mounted to be movable vertically from/to the holder 111 (stage 101), as in
The luminescence measurement unit 120a, as depicted in
The structure provided as above allows to press the luminescence measurement unit 120a more tightly against the microplate M than in the first embodiment. Such a structure may improve adhesion between the microplate M and the luminescence measurement unit 120a and prevent a crosstalk caused by light from an adjacent well.
Moreover, the chemiluminescence measurement apparatus 1 is provided with the plate mask 400 as depicted in
Microplates M have minute unevenness that differs depending on standard specifications and manufacturers. Consequently, some of the microplates M poses a problem that adhesion may be deteriorated when pressing the photosensitive surface of the photosensitive portion against the microplate M. The deterioration of adhesion brings about a problem of a crosstalk caused by the light from the well W adjacent to the target well for luminescence measurement.
To cope with such a problem, in the second embodiment, the microplate M is placed thereon with the plate mask 400 having the rubber portion 403 contacting the microplate M. Thus, the rubber portion 403 contacting the microplate M absorbs the difference in unevenness of the microplate M. Moreover, a surface of the metal portion 402 not provided with the rubber portion (the surface abutting the luminescence measurement unit 120) is flat. By placing such a plate mask 400 onto the microplate M, the surface that the luminescence measurement unit 120a contacts can be flat. Moreover, the soft rubber portion 403 allows to improve adhesion when the luminescence measurement unit 120a is pressed against the microplate. Then, this improvement of adhesion allows to prevent the crosstalk caused by the light from the well W adjacent to the target well for luminescence measurement.
Furthermore, use of the luminescence measurement unit 120a provided with the rubber portion 124 around the photosensitive portion 122 together with the plate mask 400 can prevent crosstalk.
Alternatively, use of only the luminescence measurement unit 120a provided with the rubber portion around the photosensitive portion 122 without using the plate mask 400 can also prevent crosstalk.
When dispensing a fixed quantity of a luminescent reagent continuously and at high speed, dripping from the tip of the nozzle 140 becomes a problem. Occurrence of dripping poses a problem that an accurate liquid volume cannot be dispensed. Also, there is a problem that a dripped sample and/or luminescent reagent remaining on the deck D (see
In the following, reference is made to
First, before dispensing a reagent into wells W, the dispensing controller 212 causes the nozzle 140 to eject a luminescent reagent 303 (S201; see
Next, after completion of ejecting the luminescent reagent 303, the dispensing controller 212 controls the cylinder (not depicted) to suck in air as much as a liquid volume q from the nozzle 140 (S202). This causes an interior of the nozzle 140 to be filled with air from the tip to a level to suck in as much as the liquid volume q, as depicted in
Then, moving the stage 101 horizontally and downward (S21) are performed by the movement controller 211 to position the nozzle 140 just above a well into which the luminescent reagent 303 is dispensed first. This process corresponds to the process of the steps S111, S112, S121, and S122 in
Next, the dispensing controller 212 performs dispensing the luminescent reagent 303 into the well W (S212). This process corresponds to the step S113 or step S123 in
When dispensing finishes, the dispensing controller 212 controls the cylinder (not depicted) to suck in air as much as the liquid volume q through the nozzle 140 (S213). This causes an interior of the nozzle 140 to be filled with air from the tip to the level of as much as the liquid volume q, as depicted in
Subsequently, the processing unit 210 repeats steps S211 through 213 until dispensing finishes.
In the third embodiment, sucking in air as much as the liquid volume q is done beforehand and the dispensing controller 212 performs the dispensing the luminescent reagent 303 of a liquid volume Q desired to be dispensed+vacant liquid volume q. After dispensing, air as much as the liquid volume q is sucked in to prevent dripping. Thus, dripping can be prevented, and an accurate liquid volume can be dispensed.
Next, a description is given of a procedure for dispensing the luminescent reagent 303 and performing the luminescence measurement in a fourth embodiment with reference to
(B1) First, dispensing the luminescent reagent 303 into the well Wa is performed by the nozzle 140, as depicted in
(B2) Next, horizontally moving the stage 101 is performed by the movement controller 211. At this time, the stage 101 is horizontally moved above the well Wc skipping the well Wb adjacent to the well Wa into which the reagent is already dispensed at (B1) so that dispensing the reagent into the well Wc may be performed. That is, dispensing the luminescent reagent 303 into the well Wc is performed by the nozzle 140, as depicted in
(B3) Then, horizontally moving the stage 101 is performed by the movement controller 211. At this time, the stage 101 is horizontally moved skipping a well Wd adjacent to the well Wd into which the reagent was dispensed in (B2) so that dispensing the reagent into a well We is performed. That is, dispensing the luminescent reagent 303 into the well We is performed through the nozzle 140, as depicted in
(B4) After several steps, dispensing the reagent into a well Wi located at a terminal side of the microplate M is performed through the nozzle 140, as depicted in
(B5) After that, a measurement is performed of luminescence produced by the mixture liquid 302 in the well Wi into which the reagent is dispensed at (B4). At this time, dispensing through the nozzle 140 is not performed.
(B6) Then, the movement controller 211 returns the nozzle 140 and the luminescence measurement unit 120 to a starting side of the microplate M. At this time, the stage 101 is horizontally moved to perform dispensing the reagent into the well Wb which is located near the starting side of the microplate M and into which the reagent is not yet dispensed, as depicted in
(B7) Then, the movement controller 211 horizontally moves the stage 101. At this time, the stage 101 is horizontally moved to perform dispensing the reagent into the well Wd, skipping the well Wc adjacent to the well Wb into which the reagent is dispensed at (B6). Note that the well Wc is already dispensed with the luminescent reagent 303at (B2). That is, dispensing the luminescent reagent 303 into the well Wd is performed through the nozzle 140, as depicted in
(B8) Then, horizontally moving the stage 101 is performed by the movement controller 211. At this time, the stage 101 is horizontally moved skipping the well We adjacent to the well Wd into which the reagent is dispensed at (B7) to perform, dispensing the reagent into a well Wf. Note that the luminescent reagent 303 is already dispensed into the well We at (B3). That is, dispensing the luminescent reagent 303 into the well Wf is performed through the nozzle 140, as depicted in
(B9) After several steps, dispensing the reagent into a well Wj located near the terminal side of the microplate M is performed through the nozzle 140, as depicted in
(B10) After that, a measurement is performed of luminescence produced by the mixture liquid 302 in the well Wi into which the reagent is dispensed in (B9). At this time, dispensing through the nozzle 140 is not performed.
As depicted in
It would be understandable that dispensing and luminescence measurement are performed every other well, as in
In
In the chemiluminescence measurement apparatus 1a depicted in
The chemiluminescence measurement apparatus 1 may be equipped with plural nozzles 140.
Further, the chemiluminescence measurement apparatus 1 is covered with a cover or the like which is not depicted, which allows to prevent crosstalk caused by light in a room.
Note that the springs 114 may be omitted in both the first and second embodiments.
Furthermore, the embodiments may be provided with plural nozzles 140 and plural luminescence measurement units 120. In this case, the number of the nozzles 140 is preferably equal to the number of the luminescence measurement units 120, which allows to perform dispensing a luminescent reagent and luminescence measurement for wells W arranged in plural lines at the same time.
Note that the present invention is not limited to the embodiments described above and includes various modifications. For example, the above-described embodiments are described in detail to explain the present invention clearly and the invention is not necessarily limited to those including all components described. Further, a subset of components of an embodiment may be replaced by one or more components of another embodiment and a subset of components of an embodiment may be added with one or more components of another embodiment. Furthermore, for a subset of the components of each embodiment, other components of the embodiment may be added thereto or replace the subset, or the subset may be deleted.
Besides, a subset or all of the aforementioned components, functions, each unit 210 to 213, or the like, may be implemented by hardware; for example, by a design of an integrated circuit. Also, the aforementioned components, functions, and the like, as depicted in
Moreover, in each of the embodiments, control lines and information lines which are considered as necessary for explanation are delineated and all control lines and information lines involved in a product are not necessarily delineated. Actually, almost all components may be considered to be interconnected.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2019-011323 | Jan 2019 | JP | national |
