JP2020043769A - Multiwell plate for model organisms - Google Patents

Abstract

To make it possible to increase the accuracy of simultaneously maintaining the postures of a plurality of model organisms in a multiwell plate for model organisms.SOLUTION: A multiwell plate for model organisms of the present invention has a plurality of wells 3 which are formed so as to be recessed from a plate top part 1 toward a plate bottom part 2, the plurality of wells 3 are capable of housing a plurality of model organisms M, a well bottom wall 6 of each well 3 has a recess 7 recessed toward the plate bottom part 2, the recesses 7 of the plurality of wells 3 are capable of housing a plurality of the model organisms M, each recess 7 includes a head corresponding area 8 and a body corresponding area 9 having peripheral surfaces 8a, 9a which are severally formed corresponding to standard shapes of a head m1 and a body part m2 of the model organism M, the plurality of recesses 7 face the same direction, and the postures of the plurality of model organisms M are determined on the basis of the peripheral surfaces of the plurality of recesses 7.SELECTED DRAWING: Figure 3

Description

  The present invention relates to a multi-well plate for model organisms configured to be capable of accommodating a plurality of model organisms in a plurality of wells.

  In the medical field, the biological field, and the like, particularly in the drug discovery process, attention has been paid to using fish such as medaka and zebrafish as model organisms for drug efficacy evaluation tests. In particular, in drug discovery screening, screening of a plurality of fishes should be carried out using a multi-well plate for model organisms having a plurality of hollow-shaped wells (hereinafter simply referred to as “multi-well plate” as necessary). Is attracting attention. Specifically, in screening using a multi-well plate for model organisms (hereinafter referred to as “multi-well plate-type screening” as necessary), a plurality of fish were individually housed in a plurality of wells. In this state, it is visually observed or photographed by an imaging device.

  In such a multi-well plate type screening, it is desirable to maintain a plurality of fish in the same posture for accurate observation. However, it is difficult to maintain a plurality of fish stored in a plurality of wells in the same posture. For this reason, a multi-well plate type screening that enables a plurality of fish to be maintained in the same posture has been proposed.

  As an example of such a multiwell plate type screening, the use of the following small fish arraying device can be mentioned. Specifically, the small fish array device includes a well array plate corresponding to a multi-well plate, and each well of the well array plate has an opening that opens on the top surface side of the well array plate; A flat bottom located on the bottom side of the well array plate so as to face and formed in a rectangular shape, and a side wall formed to taper from the periphery of the opening toward the periphery of the flat bottom, The flat bottoms of the plurality of wells are oriented so as to face in the same direction as each other, and each small fish is configured to be urged by the urging means to the flat bottom of the well containing the small fish. Then, such an urging means includes a water suction pump for sucking liquid in the well from a hole formed near the flat bottom, a vibrating device for vibrating the well array plate, and a direction from the top surface of the well array plate to the bottom surface. A rotating body that rotates the well array plate so as to generate a centrifugal force, or a permanent magnet that can magnetically attract a magnetic substance contained in the body of a small fish. (For example, see Patent Document 1)

JP 2014-169951 A

  However, in an example of the multiwell plate type screening described above, the small fish can be maintained in a posture in which the body length direction is aligned with the long side direction of the rectangular flat bottom portion. There is a possibility that a part of the head faces one side in the long side direction of the flat bottom and the head of the remaining part of the plurality of small fishes faces the other side in the long side direction of the flat bottom. There is a possibility that the back fin of a part of the fish faces one side in the short side direction of the flat bottom, and the back fin of the remaining part of the plurality of small fish faces the other side of the short side of the flat bottom. That is, it is difficult to determine the same orientation of the head of the small fish, and it is also difficult to determine the same orientation of the back fin of the small fish.

  Furthermore, in an example of the above-described multi-well plate type screening, since the small fish is urged by the urging means, the equipment for performing the screening may be complicated or large.

  In view of such a situation, in a multi-well plate for model organisms, it is desired to improve the accuracy of maintaining the same posture of a plurality of model organisms. Furthermore, optionally, it is desired to prevent equipment for performing screening using a multi-well plate for model organisms from becoming complicated or large, and to perform high-throughput screening with a small amount of drug.

  In order to solve the above problem, the multi-well plate for model organisms according to one aspect is a plate bottom arranged along the horizontal direction, from a plate bottom opposed to the plate top in a direction intersecting the horizontal direction. A plurality of wells formed so as to be depressed toward each other, each well having a well opening and a well bottom wall located near the plate top and the plate bottom, respectively, and the plurality of wells each having a plurality of model organisms. A multi-well plate for model organisms, wherein the well bottom walls of the plurality of wells are respectively formed so as to be recessed toward the plate bottom, and the plurality of wells are respectively formed. Having a plurality of recesses configured to be capable of accommodating the model organism, and A plurality of models including a head corresponding area and a body corresponding area each having a peripheral surface formed corresponding to a standard shape of a head and a body, wherein the plurality of recesses are oriented so as to face the same direction; The posture of the creature is determined based on the peripheral surfaces of the plurality of recesses.

  According to the multi-well plate according to one aspect, the accuracy of maintaining the same posture of a plurality of model organisms can be increased. Also, optionally, it is possible to prevent equipment for performing screening using the multi-well plate for model organisms from becoming complicated or large, and to perform high-throughput screening with a small amount of drug.

It is a perspective view showing roughly the multi-well plate for model organisms concerning a 1st embodiment. It is a top view showing roughly the multi-well plate for model organisms concerning a 1st embodiment. FIG. 3 is an enlarged view showing part A of FIG. 2 in a state where medaka is accommodated in one well. FIG. 4 is a sectional view taken along line BB of FIG. 3.

  The multi-well plate for model organisms according to the first and second embodiments will be described below. As an example of a model organism that can be accommodated in a multi-well plate for model organisms (hereinafter, simply referred to as “multi-well plate” as necessary) according to each embodiment, medaka (South medaka, scientific name: Oryzias latipes) is cited. Explain. However, the model organisms that can be accommodated in the multiwell plate according to each embodiment include fish other than medaka (preferably, small fish), nematodes (scientific name: Caenorhabditis elegans), and larvae (ie, tadpoles). Xenopus laevis (scientific name: Xenopus laevis). For example, fish other than medaka include other medaka species, zebrafish (scientific name: Danio rerio), platy (scientific name: Xiphophorus maculatus), swordtail (scientific name: Xiphophorus hellerii), torafugu (scientific name: Takifugu rubripes), and midrifugu (scientific name: Tetraodon nigroviridis). These living organisms can be preferably used because they enable screening as living organisms themselves, are easy to observe and handle, and can avoid ethical problems. Further, the multi-well plate according to each embodiment is preferably a disposable type, but this does not limit the present invention.

  1 and 2 used in the following description, the arrow P indicates the proximal side of the worker using the multi-well plate, and the arrow D indicates the distal side of the worker. However, the present invention is not limited to this, and the operator may use the multi-well plate in a state where the multi-well plate is located in a direction other than the direction indicated by the arrow P with respect to the multi-well plate.

[First Embodiment]
The multi-well plate according to the first embodiment will be described.

[About multi-well plate]
The outline of the multi-well plate will be described with reference to FIGS. As shown in FIGS. 1 and 2, the multi-well plate has a plurality of wells 3 formed so as to be recessed from a plate top 1 toward a plate bottom 2. Here, the plate top 1 and the plate bottom 2 face each other in a direction intersecting with the horizontal direction (hereinafter referred to as “intersecting direction”), and the plate top 1 is arranged along the horizontal direction. In addition, one side in the first direction of the horizontal direction corresponds to the proximal side of the worker indicated by the arrow P in FIGS. 1 and 2, and the other side in the first direction corresponds to FIGS. 2 is defined as corresponding to the distal side of the worker indicated by arrow D. The cross direction (indicated by an arrow C in FIGS. 1 and 2) may be substantially orthogonal to the horizontal direction.

  Referring to FIGS. 3 and 4, each of the plurality of wells 3 intersects with a well opening 4 located near the plate top 1 and a well peripheral wall 5 extending from the periphery of the well opening 4 toward the plate bottom 2. And a well bottom wall 6 located near the plate bottom 2 so as to face the well opening 4 in the direction. The well peripheral wall 5 extends between the periphery of the well opening 4 and the periphery of the well bottom wall 6. Each of the plurality of wells 3 is configured to be capable of accommodating a plurality of model organisms M, that is, a plurality of medaka M (shown in FIG. 3), one per well.

  Such a multi-well plate is preferably manufactured using a resin material, a glass material, a metal material, or the like. For example, as the resin material, polyethylene, polypropylene, polystyrene, ABS resin, polycarbonate, polyamide, PMMA, or the like can be used, but is not limited thereto. It is preferable that these materials do not include a substance that emits fluorescence. In particular, in consideration of enabling observation from the well bottom wall 6, the well bottom wall 6 is preferably substantially transparent. Further, the multi-well plate may be substantially transparent. However, depending on the application of the multi-well plate, the multi-well plate may be partially colored, or the entire multi-well plate may be colored.

  As shown in FIGS. 1 and 2 again, the outer shape of the multi-well plate, in particular, the outer shape of the plate top 1 of the multi-well plate may be formed in a substantially square shape in plan view. In this case, the substantially quadrangular outer shape is formed by a pair of first sides along the first direction in the horizontal direction and a second direction substantially orthogonal to the first direction in the horizontal direction. And a pair of second sides.

  However, the present invention is not limited to this, and the outline of the multi-well plate may be as follows. The crossing direction may cross diagonally without being substantially orthogonal to the horizontal direction. In this case, each well may be recessed in a direction oblique to the horizontal direction. Also, the multi-well plate may be substantially non-transparent, ie, the multi-well plate may be translucent or opaque. For example, the well bottom wall of the well of the multi-well plate can be made transparent, and the portion other than the well bottom wall of the well can be made opaque. The well bottom wall of the multi-well plate may be made transparent and the well peripheral wall may be made a color that can prevent light transmission, for example, white or black. The outer shape of the multi-well plate or the top of the plate may be formed in a shape other than a substantially square shape in plan view. In addition, some colors of the plurality of wells may be different from other colors of the plurality of wells.

[About well details]
Next, the well 3 will be described in detail with reference to FIGS. As shown in FIGS. 3 and 4, each of the plurality of recesses 7 is formed so as to accommodate a plurality of medaka M. In particular, each recess 7 is formed so as to be able to accommodate one medaka M.

  In the present embodiment, as an example, the site of the medaka M, which is a model organism, may be defined as follows, but this does not limit the definition of the site of the model organism. As shown in FIG. 4, the head m1 of the medaka M may be defined as a portion from the end on the mouth (not shown) side to the gill (not shown) in the body length direction. The torso m2 of the medaka M may be defined as a portion for accommodating internal organs in the body length direction. Specifically, the torso m2 of the medaka M may be defined as a portion from the gill (not shown) to the anus (not shown) in the body length direction. The tail m3 of the medaka M may be defined as a portion from the anus (not shown) to the tail fin end in the body length direction.

  Furthermore, the abdomen of the medaka M may be defined as an anal edge in the body height direction. The back of the medaka M may be defined as an edge on the back fin side in the body height direction. The side surface portion m4 of the medaka M may be defined as a portion between the abdomen and the back in the body height direction. Each of the abdomen, side part m4, and back of the medaka M may include the head m1, the body m2, and the tail m3 of the medaka M, and conversely, the head m1, the body m2 of the medaka M. , And tail m3 may include the abdomen, side m4, and back of medaka M.

  The definition of the medaka M site is applicable to fish other than medaka M and model organisms such as Xenopus pre-larvae. Furthermore, when the model organism is a nematode, the head of the nematode may be defined as a portion from the mouth end to the pharynx in the body length direction. The body part of a nematode may be defined as a part for accommodating an internal organ in a body length direction. Specifically, the body of the nematode may be defined as a portion from the pharynx to the anus in the body length direction. The tail of a nematode may be defined as the portion from the anus to the tail end in the body length direction.

  Furthermore, the abdomen of the nematode may be defined as a marginal portion on the anal side in the body height direction. The back of the nematode may be defined as a portion facing the abdomen in the body height direction. The side part of the nematode may be defined as a part between the abdomen and the back in the body height direction. Each of the abdomen, side, and back of the nematode may include the head, body, and tail of the nematode, and conversely, each of the head, body, and tail of the nematode , The abdomen, side, and back of the nematode.

  The well bottom walls 6 of the plurality of wells 3 each have a plurality of recesses 7 formed so as to be recessed toward the plate bottom 2. Each recess 7 is opposed to the recess opening 7a in a crossing direction with a recess opening 7a located near the plate top 1, a recess peripheral surface 7b extending from the peripheral edge of the recess opening 7a toward the plate bottom 2. And a concave bottom surface 7c located near the plate bottom 2. The concave peripheral surface 7b extends between the peripheral edge of the concave opening 7a and the peripheral edge of the concave bottom surface 7c. The concave peripheral surface 7b may be tapered so that the area of the cross section increases from the concave bottom surface 7c toward the concave opening 7a.

  Each recess 7 has a head corresponding area 8 configured to accommodate the head m1 of the medaka M and a body corresponding area 9 configured to accommodate the body m2 of the medaka M. Further, each recess 7 may have a tail corresponding area 10 configured to accommodate the tail m3 of the medaka M.

  The peripheral surface 7b of each recess 7 is formed corresponding to the standard shape of the medaka M. More specifically, the head corresponding area 8 forms a part of the recessed peripheral surface 7b, and is formed to correspond to the standard shape of the head m1 of the medaka M (hereinafter, as required, , “Head-corresponding peripheral surface”) 8a. The torso portion corresponding area 9 constitutes a part of the concave portion peripheral surface 7b and has a circumferential surface formed corresponding to the standard shape of the torso portion m2 of the medaka M (hereinafter referred to as “the torso corresponding peripheral portion” as necessary). Surface 9a). The tail-corresponding area 10 constitutes a part of the concave peripheral surface 7b and is formed in accordance with the standard shape of the tail m3 of the medaka M (hereinafter referred to as "tail-corresponding peripheral" as necessary). ) 10a. The details of the “standard shape” will be described later.

  Each of the recesses 7 may be formed so that the medaka M accommodated in the recess 7 is oriented in such a manner that the side surface m4 of the medaka M faces the plate top 1 (hereinafter, referred to as “lateral orientation”). In this case, the head-corresponding peripheral surface 8a is formed corresponding to the standard shape of the head m1 of the medaka M in the lateral position, and the trunk corresponding peripheral surface 9a is formed of the trunk m2 of the medaka M in the lateral posture. The tail corresponding peripheral surface 10a is formed corresponding to the standard shape, and is formed corresponding to the tail m3 of the medaka M in the horizontal posture.

  The plurality of recesses 7 are oriented so as to face the same direction. In a state where the plurality of medakas M are accommodated in the plurality of recesses 7, respectively, the postures of the plurality of medakas M are determined to be the same based on the peripheral surfaces 7b of the plurality of recesses 7, respectively.

  In the well 3 of the multi-well plate according to the present embodiment, the periphery of the well opening 4, the cross section of the well peripheral wall 5, and the periphery of the well bottom wall 6 are each formed in a substantially square shape. In this case, the concave portion 7 is preferably arranged along a diagonal line of the well bottom wall 6 formed in a substantially square shape. That is, the concave portion 7 may be arranged so that the longitudinal direction along the body length direction of the medaka M is directed obliquely to the first or second direction in the horizontal direction. Each well bottom wall 6 may have a bottom wall base surface 11 located near the plate top 1. In this case, the recess 7 is formed so as to be recessed from the bottom wall base surface 11 toward the plate bottom 2. The bottom wall base surface 11 is preferably formed substantially flat.

  The recess 7 may be formed so as to be capable of accommodating the entire medaka M. The peripheral surface 7b of the concave portion 7 is preferably formed so that a gap is formed between the peripheral surface 7b and the medaka M accommodated in the concave portion 7. The distance of this gap is preferably about 5 μm to about 10 μm. The recess 7 may also be asymmetrical in the longitudinal direction corresponding to the length direction of the medaka M. The recess 7 may be asymmetric in a direction substantially orthogonal to the longitudinal direction of the recess 7 in the horizontal direction. The shape of the peripheral surface 7b of the concave portion 7 formed in each of the plurality of wells included in one multi-well plate may be the same. It is preferable that the peripheral surface 7b of the concave portion 7 is as close as possible to the well peripheral wall 5. For example, it is preferable that the end area of the peripheral surface 7b in the longitudinal direction of the concave portion 7 be arranged so as to substantially coincide with the well peripheral wall 5.

  However, the shape of the well of the present invention is not limited to the above, and can be formed as follows. At least one of the peripheral portion of the well opening, the cross section of the peripheral wall of the well, and the peripheral portion of the bottom wall of the well may be formed in a shape other than a substantially square shape. For example, the peripheral portion of the well opening and the peripheral portion of the well peripheral wall may be formed. At least one of the cross section and the peripheral edge of the well bottom wall can be formed in a substantially circular shape, a substantially elliptical shape, a substantially polygonal shape other than a substantially square shape, or the like. The recess may be arranged so that the longitudinal direction along the body length direction of the medaka is directed to the first or second direction of the horizontal direction. The bottom wall base surface of the well bottom wall can be formed in a shape other than a substantially flat shape. For example, the bottom wall base surface may be formed in a substantially curved shape, a substantially bent shape, a substantially conical shape, a substantially pyramid shape, or the like. The well bottom wall may be configured to have no bottom wall base surface. Further, depending on the use of the multi-well plate, a plurality of wells in one multi-well plate have a plurality of recesses respectively formed in different shapes based on a standard shape having a different growth stage described later. You may.

  Further, the arrangement of the well 3 is preferably as follows. As shown in FIGS. 1 and 2, the plurality of wells 3 are arranged in a matrix along a plurality of rows and a plurality of columns in the horizontal direction. As shown in FIG. 2, with respect to the orientation direction of the matrix, a plurality of rows are arranged along a second direction orthogonal to the first direction among the horizontal directions, and a plurality of columns are arranged in the horizontal direction. Are arranged along the first direction.

  In the present embodiment, the plurality of wells 3 are arranged in a matrix having 16 rows and 24 columns in the horizontal direction, and 24 wells 3 are arranged in each row, and each column has Sixteen wells 3 are arranged. The multi-well plate has 384 wells 3.

  However, the arrangement of the wells of the present invention is not limited to this, and may be as follows. A plurality of wells can be arranged in a matrix having n rows and m columns in the horizontal direction, m wells can be arranged in each row, and n wells can be arranged in each column. Can be arranged. In this case, the multi-well plate has (n × m) wells. Note that each of n and m is preferably an integer of 2 or more, particularly an integer of 3 or more. For example, the number of wells arranged in a matrix can be 6, 12, 24, 48, 96, 1536, or the like. As a specific example, in a multi-well plate, 96 wells can be arranged in a matrix having 8 rows and 12 columns in the horizontal direction.

  Further, the plurality of wells can be arranged in a state other than the matrix. For example, the number of wells arranged along a part of the plurality of rows may be different from the number of wells arranged along another part of the plurality of rows. Regarding the orientation direction of the matrix regarding the well, if the orientation directions of the plurality of rows and the orientation directions of the plurality of columns are substantially orthogonal to each other, the orientation direction of the plurality of rows is set to a direction other than the second direction, and May be set to a direction other than the first direction.

[Details of standard shape]
The details of the above-described standard shape will be described with reference to FIG. The standard shape of the medaka M may be an average shape of the medaka M calculated based on a result of previously measuring the shapes of a plurality of samples related to the medaka M. It is generally known that medaka has many inbred lines whose genes are substantially uniform, and that individual differences in the shape of embryos at the developmental stage of each inbred line are small. Specifically, the standard shape of the head m1, the body m2, and the tail m3 of the medaka M is a plurality of samples of the head m1, the body m2, and the tail m3, for example, about 3 to about 5 animals. The average shape of the head m1, the body m2, and the tail m3 of the medaka M, which is calculated based on the result of measuring the shape of the sample in advance, may be used. Further, the standard shapes of the medaka M, the head m1, the body m2, and the tail m3 can be determined according to the stage at which the medaka M is generated. The development stage may be an embryo stage or a fry stage after a predetermined number of days have passed since fertilization. Therefore, for example, the standard shape of the embryo on the 5th day after fertilization, the standard shape of the embryo on the 6th day after fertilization, and the like are calculated, and a multiwell plate for observation of the embryo on the 5th day after fertilization, A multi-well plate or the like for observation can be separately prepared.

  For example, such an average shape can be calculated by manual calculation or automatic calculation based on dimensions related to the shape of a plurality of samples measured using a dimension measuring device such as a ruler. For example, the average shape can also be calculated by analyzing image data on a plurality of samples acquired using an image acquisition device such as a laser scanner or a camera.

  As described above, according to the multi-well plate according to the present embodiment, the medaka M accommodated in the concave portion 7 of each well 3 is such that the head m1 and the body m2 of the medaka M correspond to the head corresponding area 8 of the concave portion 7, respectively. The plurality of recesses 7 are arranged so as to correspond to the body corresponding area 9 and are oriented so as to face the same direction. Therefore, it is possible to improve the accuracy of maintaining the same posture of the plurality of medakas M in the multi-well plate. Further, when screening using a multi-well plate is performed, a medaka M in each well 3 is filled with a drug solution in each well 3 so that the head of each medaka M is formed in the recess 7. m1 and the torso m2 are guided to be arranged in the head corresponding area 8 and the torso corresponding area 9. It is preferable that the amount of the drug solution supplied at this time is an amount corresponding to the volume of each well and determined so that the medaka M can be guided to the concave portion 7. For example, the amount of such a drug solution may have a minimum volume of about 40 μL or less, and preferably 20 μL or less. Further, for example, the amount of the drug solution is preferably about 10 μL or more. However, the amount of the drug solution is not limited to these. In the case where medaka that has already been treated with a drug is injected into the well and used, a buffer salt solution or the like having a similar amount may be injected into the well 3 instead of the drug solution. Therefore, the posture of the medaka M can be maintained in the well 3 without using the conventional urging means, so that the equipment for performing the screening using the multi-well plate becomes complicated or large. It is possible to perform high-throughput screening with a small amount of drug.

  According to the multi-well plate according to the present embodiment, the standard shape of medaka M is an average shape calculated based on the results of measuring the shapes of a plurality of samples related to medaka M. Therefore, the concave portions 7 of the plurality of wells 3 formed based on the average shape improve the accuracy of maintaining the plurality of medakas M in the same posture without being affected by individual differences of the medakas M. Can be.

  According to the multi-well plate according to the present embodiment, the concave portion 7 of each well 3 further includes the tail corresponding area 10 having the peripheral surface 10a formed corresponding to the standard shape of the tail m3 of the medaka M. Therefore, the plurality of recesses 7 formed corresponding to the standard shapes of the head m1, the body m2, and the tail m3 of the medaka M can improve the accuracy of maintaining the plurality of medaka M in the same posture. .

  According to the multi-well plate according to the present embodiment, the concave portion 7 of each well 3 defines the medaka M accommodated in the concave portion 7 such that the side surface m4 of the medaka M faces the plate top 1. Is formed. Therefore, it is possible to increase the accuracy of maintaining the plurality of medakas M in the same posture with their side surfaces m4 facing the plate top 1.

[Second embodiment]
A multi-well plate according to the second embodiment will be described. The multi-well plate according to the present embodiment has the same configuration as the multi-well plate according to the first embodiment, except for the following points.

  Although not particularly shown, in the multi-well plate according to the present embodiment, each of the concave portions is configured such that the medaka accommodated in the concave portion has a posture in which the back portion of the medaka is directed toward the top of the plate (hereinafter, the “prone position”). Medaka) or a posture in which the abdomen of the medaka is directed toward the top of the plate (hereinafter, referred to as a “supine posture”). In this case, the head-corresponding peripheral surface is formed corresponding to the standard shape of the medaka head in the prone or supine position, and the torso-corresponding peripheral surface is the medaka torso in the prone or supine position. And the tail-corresponding peripheral surface is formed corresponding to the tail of the medaka in the prone or supine position.

  As described above, according to the multi-well plate according to the present embodiment, the recess 7 of each well 3 causes the medaka M accommodated in the recess 7 to face the side m4 of the medaka M toward the top 1 of the plate. Except for the effect based on the configuration formed as defined, the same effect as the multi-well plate according to the first embodiment can be obtained.

  Further, according to the multi-well plate according to the present embodiment, the recess of each well is formed so as to determine the medaka accommodated in the recess in a posture in which the back or the abdomen of the medaka is directed toward the top of the plate. Therefore, the accuracy of maintaining the plurality of medakas in the same posture with their back or abdomen facing the top of the plate can be enhanced.

[How to use multi-well plate]
Here, a method of using the multi-well plate according to each of the first and second embodiments will be briefly described. The method of using the multi-well plate includes a step of injecting the model organism into the multi-well plate, and a step of acquiring an image of the model organism. According to the multi-well plate according to each of the first and second embodiments, the model organisms are aligned without any special operation due to the structural characteristics, and image acquisition becomes easy. The growth stage (day after fertilization) of the model organism to be injected is not particularly limited, and it can be injected at the stage of a fertilized egg or at the stage after hatching. The model organism to be injected may have already been treated with the candidate drug to be screened. Alternatively, at the same time as or after the step of injecting the model organism into the multiwell plate, a compound and / or a candidate drug having a different concentration can be injected into each well, and drug treatment can be performed in the wells. The injection of the model organism and / or the candidate drug can be performed using an automatic dispensing device, but the injection method is not particularly limited. Although the growth stage of the model organism at the time of image acquisition is not particularly limited, if the shape of the concave portion 7 of the multi-well plate is designed so as to conform to the predetermined growth stage, the predetermined growth stage It is preferable to acquire an image. This is because a higher alignment ratio can be obtained.

  According to such a method of use, the steps from drug treatment of the model organism to image acquisition can be performed in one multi-well plate, and an operation for aligning the model organism is not required. Therefore, high-throughput screening becomes possible. Note that, in one embodiment, the present invention is a drug screening method, wherein a step of contacting a candidate drug with a model organism, and a step of injecting the model organism into the multi-well plate according to the first and second embodiments, Obtaining an image of the model organism. The step of bringing the candidate agent into contact with the model organism may be performed before, after, or simultaneously with the step of injecting the model organism into the multiwell plate.

  Although the embodiments of the present invention have been described so far, the present invention is not limited to the above-described embodiments, and the present invention can be modified and changed based on the technical idea.

  An example will be described. In this example, a multi-well plate according to the first embodiment having 384 wells 3 was manufactured. Such a multiwell plate was substantially the same as that shown in FIG. Using this multi-well plate, medaka was phenotype-screened for candidate drugs. Two or three days after fertilization, embryos at the stage of embryos are fed one by one to one well, and about 20 μL of a candidate drug or a candidate drug-containing solution is injected into each well to perform drug treatment. did. Thereafter, an image of medaka M at the stage of embryo 3 days after the drug treatment was obtained.

  Specifically, first, medaka eggs at 2 to 3 days after fertilization and at the stage of an embryo having substantially the same growth stage are rubbed with sandpaper, and then the eggshell is processed by hatching enzyme treatment. Removed. Next, the medaka embryo from which the eggshell has been removed is washed twice or three times with a buffered saline solution, and then the multiwell plate is placed with its plate top 1 and plate bottom 2 facing upward and downward, respectively, using a dropper. Then, together with about 10 μL of the buffered saline, one medaka egg per well was placed. Further, about 384 kinds of drug solutions having different compositions were injected into each of the 384 wells 3 by about 10 μL per well to carry out drug treatment. Thereafter, an anesthetic (MS222) was added to the medaka M at the stage of the embryo three days after the drug treatment so that the final concentration was about 0.04%, and about 5 minutes to about 10 minutes. Later, it was confirmed that Medaka M was anesthetized. At this time, the medaka M in each well 3 had substantially the same posture due to the shape of the recess 7 of the well 3. On the other hand, when there is a medaka M having a posture different from the shape of the concave portion 7, a straight needle is bent at approximately a middle portion of the medaka M at approximately 45 ° to approximately 60 ° with respect to the medaka M. The medaka M was prepared using the needle handle formed in the same manner as that of the other medaka M. Once the medaka M in each well 3 in which the posture has been adjusted is supported by the shape in the concave portion 7, the medaka M is kept in the substantially same posture unless the medaka M is moved by the release of anesthesia. Maintained.

  Further, using an image analyzer, Opera Phenix manufactured by PerkinElmer, images of 384 medaka M contained in the multiwell plate were obtained. As a result, an image in which all the 384 medaka M are arranged in a matrix having 16 rows and 24 columns and arranged in the same posture with each other is acquired in a short time of less than about 10 minutes. And good screening could be performed using such images. In particular, in the obtained image, 384 medaka M are captured in the same posture, and an enlarged image of a desired portion can be obtained. Therefore, for example, in drug screening using a transgenic medaka in which a specific organ is visualized with a fluorescent protein, it becomes easy to obtain an enlarged image of only the specific organ.

DESCRIPTION OF SYMBOLS 1 ... Plate top part, 2 ... Plate bottom part, 3 ... Well, 4 ... Well opening, 6 ... Well bottom wall 7 ... Concave part, 7b ... Concave peripheral surface, 8 ... Head corresponding area, 8a ... Peripheral surface (Head corresponding peripheral) 9) ... peripheral surface (body-corresponding peripheral surface), 10 ... tail-corresponding area, 10a ... peripheral surface (tail-corresponding peripheral surface), M ... medaka (model organism), m1 ... head Part, m2 ... body part, m3 ... tail part

In order to solve the above problem, the multi-well plate for model organisms according to one aspect is a plate bottom arranged along the horizontal direction, from a plate bottom opposed to the plate top in a direction intersecting the horizontal direction. toward comprising a plurality of wells formed as recessed, each well has a respective said plate top portion and the plate bottom near the well opening and the well bottom wall located in said plurality of wells, multiple model organisms the is configured to be accommodated respectively, a multi-well plate for model organisms, well bottom walls of each well has a recess formed so as to be recessed toward the plate bottom, the plurality recess of the well wall in the well is configured of the plurality of model organisms so as to accommodate each of the recesses, the recess Having a head corresponding areas with the peripheral surface which is formed to correspond to the standard shape of the head of the model organisms that are accommodated, the peripheral surface is formed corresponding to the standard shape of the body portion of the model organism and a body portion corresponding region, wherein the plurality of recesses are aligned so that the same direction, the orientation of the plurality of model organisms, so that the respective determined based on the circumferential surface of the plurality of recesses .

The well bottom wall 6 of each well 3 has a recess 7 formed so as to be recessed toward the plate bottom 2. Each recess 7 includes a recess opening 7a located Plate top 1 close, and the recess inner peripheral surface 7b extending from the peripheral edge of the recess opening 7a to the plate bottom 2, so as to face the recess opening 7a at the cross direction And a concave bottom surface 7c located near the plate bottom 2. The concave peripheral surface 7b extends between the peripheral edge of the concave opening 7a and the peripheral edge of the concave bottom surface 7c. The concave peripheral surface 7b may be tapered so that the area of the cross section increases from the concave bottom surface 7c toward the concave opening 7a.

However, the shape of the well of the present invention is not limited to the above, and can be formed as follows. At least one of the peripheral portion of the well opening, the cross section of the peripheral wall of the well, and the peripheral portion of the bottom wall of the well may be formed in a shape other than a substantially square shape. For example, the peripheral portion of the well opening and the peripheral portion of the well peripheral wall may be formed. At least one of the cross section and the peripheral edge of the well bottom wall can be formed in a substantially circular shape, a substantially elliptical shape, a substantially polygonal shape other than a substantially square shape, or the like. The recess may be arranged so that the longitudinal direction along the body length direction of the medaka is directed to the first or second direction of the horizontal direction. The bottom wall base surface of the well bottom wall can be formed in a shape other than a substantially flat shape. For example, the bottom wall base surface may be formed in a substantially curved shape, a substantially bent shape, a substantially conical shape, a substantially pyramid shape, or the like. The well bottom wall may be configured to have no bottom wall base surface. Further, depending on the use of the multi-well plate, each of the plurality of wells in one multi-well plate has a concave portion, and the concave portions of the plurality of wells have different shapes based on a standard shape having a different growth stage described later. May be formed respectively.

Claims (5)

From the top of the plate arranged along the horizontal direction, comprising a plurality of wells formed so as to be depressed toward the bottom of the plate facing the top of the plate in a direction intersecting with the horizontal direction,
Each well has a well opening and a well bottom wall located near the plate top and the plate bottom, respectively.
The plurality of wells are configured to be capable of accommodating a plurality of model organisms, respectively, a model organism multi-well plate,
Well bottom walls of the plurality of wells each have a plurality of recesses formed to be recessed toward the plate bottom and configured to be capable of accommodating the plurality of model organisms, respectively.
Each recess includes a head corresponding area and a body corresponding area each having a peripheral surface formed corresponding to the standard shape of the head and the body of the model organism,
The plurality of recesses are oriented so as to face the same direction,
A multi-well plate for model organisms, wherein postures of the plurality of model organisms are determined based on peripheral surfaces of the plurality of recesses, respectively.   The multi-well plate for a model organism according to claim 1, wherein the standard shape is an average shape calculated based on a result of measuring shapes of a plurality of samples related to the model organism. The model organism is a fish,
2. The multi-well plate for model organisms according to claim 1, wherein each recess further includes a tail corresponding area having a peripheral surface formed corresponding to a standard shape of the fish tail.   4. The multi-well plate for a model organism according to claim 3, wherein each recess is formed so as to determine the fish accommodated in the recess in a posture in which a side portion of the fish faces the top of the plate. 5.   The multi-well plate for a model organism according to claim 3, wherein each recess is formed so as to determine the fish accommodated in the recess in a posture in which the back or the abdomen of the fish faces the top of the plate. .

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