JP6560054B2 - Specimen introduction member, specimen introduction method to well - Google Patents
Specimen introduction member, specimen introduction method to well Download PDFInfo
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- JP6560054B2 JP6560054B2 JP2015155464A JP2015155464A JP6560054B2 JP 6560054 B2 JP6560054 B2 JP 6560054B2 JP 2015155464 A JP2015155464 A JP 2015155464A JP 2015155464 A JP2015155464 A JP 2015155464A JP 6560054 B2 JP6560054 B2 JP 6560054B2
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- 230000002209 hydrophobic effect Effects 0.000 claims description 43
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- 238000007847 digital PCR Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
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- 108020004707 nucleic acids Proteins 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 229920002379 silicone rubber Polymers 0.000 description 2
- 239000004945 silicone rubber Substances 0.000 description 2
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- 238000003752 polymerase chain reaction Methods 0.000 description 1
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- 238000011002 quantification Methods 0.000 description 1
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Description
本発明は、ウェルへ検体を導入する際に使用される検体導入部材およびこれを用いたウェルへの検体導入方法に関するものである。 The present invention relates to a sample introduction member used when introducing a sample into a well and a method for introducing a sample into a well using the same.
デジタルＰＣＲ（Ｐｏｌｙｍｅｒａｓｅ Ｃｈａｉｎ Ｒｅａｃｔｉｏｎ）法は、ごく微量のＤＮＡやＲＮＡを短時間で増幅する方法として知られている。デジタルＰＣＲは、検体をプライマ、試薬等とともに（以下略して単に検体とする。）微細なマイクロウェルに導入し、遺伝子増幅した後に、蛍光などでターゲット遺伝子を検知して、元の検体の核酸数を定量化しようとするものである。 Digital PCR (Polymerase Chain Reaction) method is known as a method for amplifying a very small amount of DNA or RNA in a short time. In digital PCR, a sample is introduced into a fine microwell together with primers, reagents, and the like (hereinafter simply referred to as a sample), amplified, and then the target gene is detected by fluorescence or the like to detect the number of nucleic acids in the original sample. Is to be quantified.
特に、近年は、複数のウェルのそれぞれに検体が入れられ、これらを順次、自動で計測することで、より多くの検体を効率良く測定する方法が要求される。このため、複数のウェルのアレイを有するマイクロウェルプレートが提案されている（例えば、特許文献１、特許文献２）。 In particular, in recent years, there is a demand for a method for efficiently measuring a larger number of specimens by placing specimens in each of a plurality of wells and sequentially and automatically measuring them. For this reason, a microwell plate having an array of a plurality of wells has been proposed (for example, Patent Document 1 and Patent Document 2).
デジタルＰＣＲで用いられるマイクロウェルプレートは、ウェルの数が多ければ多いほど検体の分割数が増加し、定量化のダイナミックレンジが向上する。一方で、同一サイズのマイクロウェルプレートに対してウェルの数を増やそうとすると、ウェルの直径が小さくなることになる。 In a microwell plate used in digital PCR, the greater the number of wells, the greater the number of sample divisions and the more the dynamic range of quantification. On the other hand, if the number of wells is increased with respect to the microwell plate having the same size, the diameter of the wells is reduced.
このような微細なウェルに検体を導入する方法としては、例えば、多数の微細なウェルが設けられたマイクロウェルプレート上に検体を滴下し、検体をへら状のものでなでることで、検体をウェル内に導入する方法が採られている。このようにして、複数のそれぞれのウェルに検体を導入し、それぞれのウェルを互いに独立した反応容器とすることで、元の検体の核酸数を定量化することができる。 As a method for introducing a specimen into such a fine well, for example, the specimen is dropped on a microwell plate provided with a large number of fine wells, and the specimen is then sprinkled with a spatula to remove the specimen from the well. The method to introduce in is taken. In this way, the number of nucleic acids in the original sample can be quantified by introducing the sample into each of a plurality of wells and making each well a reaction container independent of each other.
一方、隣り合うウェル内の検体がマイクロウェルプレート上でつながってしまうと、正確な測定ができなくなる。このため、通常、ウェル部分を除くマイクロウェルプレートの表面には、疎水化処理を施す必要がある。 On the other hand, if the samples in adjacent wells are connected on the microwell plate, accurate measurement cannot be performed. For this reason, it is usually necessary to subject the surface of the microwell plate excluding the well portion to a hydrophobic treatment.
しかし、マイクロウェルプレートの表面に疎水化処理が施されていると、検体を滴下した際に、検体の液滴がマイクロウェルプレート上で玉状になりやすく、検体をマイクロウェルプレートの全体に行きわたらせることが困難である。このため、前述した様なヘラ状の部材を用いても、マイクロウェルプレートの全ウェルへ検体を導入することが困難であった。 However, if the surface of the microwell plate has been hydrophobized, when the sample is dropped, the sample droplet tends to be ball-shaped on the microwell plate, and the sample goes to the entire microwell plate. Difficult to pass. For this reason, it is difficult to introduce the specimen into all the wells of the microwell plate even if the spatula-shaped member as described above is used.
本発明はこのような問題に鑑みてなされたもので、検体を効率良くウェルに導入することが可能な検体導入部材およびこれを用いたウェルへの検体導入方法を提供することを目的とする。 The present invention has been made in view of such problems, and an object of the present invention is to provide a sample introduction member capable of efficiently introducing a sample into a well and a method for introducing a sample into a well using the same.
前述した目的を達するために第１の発明は、マイクロウェルプレートに形成されたウェルへ検体を導入するための導入部材であって、前記導入部材は板状であり、前記導入部材の少なくとも一方の面が、前記検体との接触面であって、前記検体との接触面の下端部近傍であって前記マイクロウェルプレートとの接触部近傍の少なくとも一部に親水部が形成され、少なくとも前記親水部の上方に疎水部が形成されることを特徴とする検体導入部材である。
In order to achieve the above object, a first invention is an introduction member for introducing a specimen into a well formed in a microwell plate, wherein the introduction member is plate-shaped, and at least one of the introduction members The surface is a contact surface with the specimen, and a hydrophilic portion is formed in at least a part near the lower end portion of the contact surface with the specimen and in the vicinity of the contact portion with the microwell plate, and at least the hydrophilic portion A specimen introduction member characterized in that a hydrophobic part is formed above .
また、第１の発明は、マイクロウェルプレートに形成されたウェルへ検体を導入するための導入部材であって、前記導入部材は板状であり、前記導入部材の少なくとも一方の面が、前記検体との接触面であって、前記検体との接触面の下端部近傍であって前記マイクロウェルプレートとの接触部近傍の少なくとも一部に親水部が形成され、前記親水部は、前記検体導入部材の全幅に形成されず、少なくとも前記親水部の幅方向の両側に疎水部が形成されてもよい。
The first invention is an introduction member for introducing a specimen into a well formed in a microwell plate, wherein the introduction member is plate-shaped, and at least one surface of the introduction member is the specimen. A hydrophilic portion is formed in at least a part of the contact surface with the microwell plate and in the vicinity of the lower end portion of the contact surface with the sample, and the hydrophilic portion is formed by the sample introduction member. The hydrophobic portion may be formed at least on both sides in the width direction of the hydrophilic portion.
前記検体導入部材の前記親水部および前記疎水部が形成された面であって、前記親水部の上方には、ディスポチップの先端を保持する保持部が設けられ、前記保持部にディスポチップを装着可能であってもよい。 A surface of the sample introduction member on which the hydrophilic part and the hydrophobic part are formed, and a holding part that holds a tip of a disposable chip is provided above the hydrophilic part, and the disposable chip is attached to the holding part It may be possible.
第１の発明によれば、導入部材の検体との接触面であって、マイクロウェルプレートとの接触部近傍に親水部が形成されるため、当該部分に検体を接触させると、検体が導入部材の親水部に対して濡れ広がり、検体を導入部材の幅方向に容易に広げることができる。また、その周囲の一部に疎水部が形成されるため、親水部を超えて検体が過剰に広がることを抑制することができる。 According to the first invention, since the hydrophilic portion is formed in the vicinity of the contact portion with the microwell plate on the contact surface of the introduction member with the sample, the sample is brought into contact with the portion when the sample is brought into contact therewith. Thus, the specimen can be easily spread in the width direction of the introduction member. Moreover, since a hydrophobic part is formed in a part of the periphery, it is possible to prevent the specimen from spreading excessively beyond the hydrophilic part.
また、疎水部が親水部の上方に形成されれば、検体が親水部の上方に対して過剰に広がり、検体を無駄にすることを抑制することができる。 Further, if the hydrophobic part is formed above the hydrophilic part, it is possible to prevent the specimen from being excessively spread above the hydrophilic part and wasting the specimen.
また、親水部を検体導入部材の全幅に形成せず、親水部の幅方向の両側に疎水部を形成することで、マイクロウェルプレート上に滴下された検体を、導入部材でなでる際に、検体が導入部材の両側方から、導入部材の後方に流れることを抑制することができる。 In addition, by forming the hydrophobic part on both sides in the width direction of the hydrophilic part without forming the hydrophilic part over the entire width of the specimen introducing member, the specimen dropped on the microwell plate can be sampled with the introducing member. Can be prevented from flowing from both sides of the introduction member to the rear of the introduction member.
また、検体導入部材の親水部および疎水部が形成された面の上方に、ディスポチップの先端を保持する保持部を設けることで、検体を保持したディスポチップの先端に、直接導入部材を装着することが可能である。 In addition, the introduction member is directly attached to the tip of the disposable chip holding the sample by providing a holding unit for holding the tip of the disposable chip above the surface of the sample introducing member where the hydrophilic portion and the hydrophobic portion are formed. It is possible.
第２の発明は、検体導入部材を用いた、マイクロウェルプレートに形成されたウェルへの検体の導入方法であって、前記検体導入部材は板状であり、前記検体導入部材の少なくとも一方の面が、検体との接触面であって、前記検体との接触面の下端部近傍であって前記マイクロウェルプレートとの接触部近傍の表面の一部に親水部が形成され、少なくとも前記親水部の上方に疎水部が形成されており、複数のウェルが形成された前記マイクロウェルプレート上に、ディスポチップで検体を滴下し、前記検体に前記検体導入部材を接触させて、前記検体を前記親水部の形成範囲に広げ、前記検体導入部材で前記マイクロウェルプレートの表面をなぞり、前記検体を、前記ウェルに導入することを特徴とするウェルへの検体導入方法である。
A second invention is a method for introducing a sample into a well formed in a microwell plate using a sample introduction member, wherein the sample introduction member is plate-shaped and at least one surface of the sample introduction member Is a contact surface with the specimen, and a hydrophilic portion is formed in a part of the surface near the lower end portion of the contact surface with the specimen and in the vicinity of the contact portion with the microwell plate, and at least the hydrophilic portion A hydrophobic part is formed on the top, a specimen is dropped with a disposable chip on the microwell plate having a plurality of wells, the specimen introduction member is brought into contact with the specimen, and the specimen is brought into contact with the hydrophilic part. The specimen introduction method is characterized in that the specimen is introduced into the well by tracing the surface of the microwell plate with the specimen introduction member.
前記検体導入部材の前記親水部および前記疎水部が形成された面であって、前記親水部の上方には、ディスポチップの先端を保持する保持部が設けられ、前記保持部にディスポチップを装着し、前記マイクロウェルプレート上に、前記ディスポチップで検体を滴下した後、前記検体導入部材で前記マイクロウェルプレートの表面をなぞり、前記検体を、前記ウェルに導入してもよい。 A surface of the sample introduction member on which the hydrophilic part and the hydrophobic part are formed, and a holding part that holds a tip of a disposable chip is provided above the hydrophilic part, and the disposable chip is attached to the holding part Then, after dropping a sample on the microwell plate with the disposable chip, the sample may be introduced into the well by tracing the surface of the microwell plate with the sample introduction member.
第２の発明によれば、親水部が形成された導入部材を用いることで、検体を導入部材の幅方向に容易に広げることができる。また、その周囲の一部に疎水部が形成されるため、親水部を超えて検体が過剰に広がることを抑制することができる。このため、容易に検体をマイクロウェルプレートの全体に行きわたらせることができる。 According to the second invention, the sample can be easily spread in the width direction of the introduction member by using the introduction member in which the hydrophilic portion is formed. Moreover, since a hydrophobic part is formed in a part of the periphery, it is possible to prevent the specimen from spreading excessively beyond the hydrophilic part. For this reason, the specimen can be easily distributed throughout the microwell plate.
また、検体導入部材の親水部および疎水部が形成された面の上方に、ディスポチップの先端を保持する保持部を設けることで、検体を保持したディスポチップの先端に、直接導入部材を装着することが可能である。このため、ディスポチップによって、マイクロウェルプレート上に検体を滴下する動作と、滴下された検体をマイクロウェルプレートの全体に広げる工程とを連続して行うことができる。 In addition, the introduction member is directly attached to the tip of the disposable chip holding the sample by providing a holding unit for holding the tip of the disposable chip above the surface of the sample introducing member where the hydrophilic portion and the hydrophobic portion are formed. It is possible. For this reason, the operation of dropping the specimen onto the microwell plate and the process of spreading the dropped specimen over the entire microwell plate can be performed continuously by the disposable chip.
本発明によれば、検体を効率良くウェルに導入することが可能な検体導入部材およびこれを用いたウェルへの検体導入方法を提供することができる。 ADVANTAGE OF THE INVENTION According to this invention, the sample introduction member which can introduce a sample into a well efficiently, and the sample introduction method to a well using the same can be provided.
以下、図面を参照しながら、本発明の実施形態について説明する。図１は、検体導入部材１を示す斜視図である。検体導入部材１は、主に、ヘラ部３、柄部７等から構成される。 Hereinafter, embodiments of the present invention will be described with reference to the drawings. FIG. 1 is a perspective view showing the sample introduction member 1. The sample introduction member 1 mainly includes a spatula portion 3, a handle portion 7, and the like.
ヘラ部３は、例えばシリコーンゴムなどの柔軟性のある樹脂製である。ヘラ部３は、略板状であり、一方の面が検体接触面５となる。なお、両方の面を検体接触面５として、表裏いずれの面でも利用できるようにしてもよい。以下の説明では、一方の面を検体接触面５として説明する。 The spatula portion 3 is made of a flexible resin such as silicone rubber. The spatula portion 3 is substantially plate-shaped, and one surface serves as the specimen contact surface 5. Both surfaces may be used as either the front or back surface as the specimen contact surface 5. In the following description, one surface will be described as the specimen contact surface 5.
ヘラ部３の上部には柄部７が接合される。柄部７は、検体導入部材１を操作するための部位である。柄部７の材質は問わないが、例えば、ある程度の剛性を有する樹脂製である。 A handle 7 is joined to the top of the spatula 3. The handle 7 is a part for operating the sample introduction member 1. The material of the handle 7 is not limited, but is made of resin having a certain degree of rigidity, for example.
図２は、ヘラ部３の正面図（検体接触面５の正面図）である。ヘラ部３の検体接触面５の所定の部位には、親水部９と疎水部１１が設けられる。親水部９は、例えば、親水コーティング剤などによって形成される。親水部９は、ヘラ部３の下端部近傍（後述するマイクロウェルプレートとの接触部近傍）の少なくとも一部に形成される。図示した例では、ヘラ部３の幅方向の全幅であって、ヘラ部３の下端から、高さＡ（例えば約０．５〜３ｍｍ程度）の範囲に親水部９が形成される。 FIG. 2 is a front view of the spatula portion 3 (front view of the specimen contact surface 5). A hydrophilic part 9 and a hydrophobic part 11 are provided in a predetermined part of the specimen contact surface 5 of the spatula part 3. The hydrophilic portion 9 is formed by, for example, a hydrophilic coating agent. The hydrophilic portion 9 is formed at least in the vicinity of the lower end portion of the spatula portion 3 (in the vicinity of a contact portion with a microwell plate described later). In the illustrated example, the hydrophilic portion 9 is formed in the range of the height A (for example, about 0.5 to 3 mm) from the lower end of the spatula portion 3 in the entire width in the width direction of the spatula portion 3.
親水部９の周囲の少なくとも一部には、疎水部１１が形成される。図示した例では、親水部９の上方に疎水部１１が形成される。なお、親水部９の上方に形成される疎水部１１は、親水部９の上方から１ｍｍ以上の範囲（高さＢ）に形成されることが望ましい。疎水部１１の高さが１ｍｍ未満であると、検体が疎水部１１を超えて、疎水部１１の上方に付着する恐れがある。なお、さらに好ましくは、疎水部１１は、親水部９の上方から２ｍｍ以上の範囲に形成されることが望ましい。 A hydrophobic portion 11 is formed on at least a part of the periphery of the hydrophilic portion 9. In the illustrated example, the hydrophobic portion 11 is formed above the hydrophilic portion 9. The hydrophobic part 11 formed above the hydrophilic part 9 is preferably formed in a range (height B) of 1 mm or more from above the hydrophilic part 9. If the height of the hydrophobic part 11 is less than 1 mm, the specimen may exceed the hydrophobic part 11 and adhere to the upper part of the hydrophobic part 11. More preferably, the hydrophobic portion 11 is desirably formed in a range of 2 mm or more from above the hydrophilic portion 9.
なお、疎水部１１は、例えば疎水コーティング剤によって形成してもよいが、シリコーンゴムのように、材質として疎水性を有するものであれば、疎水化処理は不要である。また、ヘラ部３の材質が親水性である場合には、親水化処理を行わずに、親水部９を残して疎水部１１に疎水化処理のみを施してもよい。 The hydrophobic portion 11 may be formed by, for example, a hydrophobic coating agent, but the hydrophobic treatment is not necessary if the material has hydrophobicity such as silicone rubber. Further, when the material of the spatula portion 3 is hydrophilic, the hydrophobic portion 11 may be subjected only to the hydrophobizing treatment without leaving the hydrophilic portion 9 without performing the hydrophilizing treatment.
次に、検体導入部材１を用いた、マイクロウェルへの検体の導入方法について説明する。図３は、マイクロウェルプレート１７上に、ディスポチップ１３によって検体１５を滴下した状態を示す図である。なお、ディスポチップ１３は、例えば、自動化された分注器に取り付けられて動作する。この場合には、まず、ディスポチップ１３を、検体貯留部（図示せず）に移動させ、検体１５をディスポチップ１３に吸引させた後、あらかじめセットされたマイクロウェルプレート１７上に移動し、所定量の検体１５をマイクロウェルプレート１７上に滴下する。 Next, a method for introducing the sample into the microwell using the sample introduction member 1 will be described. FIG. 3 is a diagram showing a state in which the specimen 15 is dropped on the microwell plate 17 by the disposable chip 13. The disposable chip 13 is attached to an automated dispenser and operates, for example. In this case, first, the disposable chip 13 is moved to a specimen storage section (not shown), the specimen 15 is aspirated by the disposable chip 13, and then moved onto the microwell plate 17 set in advance. A fixed amount of sample 15 is dropped on the microwell plate 17.
図３に示すように、マイクロウェルプレート１７の表面には、複数のウェル１９が形成される。なお、図示した例では、ウェル１９は有底穴である例を示すが、貫通孔であってもよい。 As shown in FIG. 3, a plurality of wells 19 are formed on the surface of the microwell plate 17. In the illustrated example, the well 19 is a bottomed hole, but may be a through hole.
ウェル１９の内面には、親水化処理２１が施されている。このため、検体１５を効率よくウェル１９内に導入することができる。 The inner surface of the well 19 is subjected to a hydrophilic treatment 21. For this reason, the specimen 15 can be efficiently introduced into the well 19.
また、マイクロウェルプレート１７の表面であって、ウェル１９以外の部位には疎水化処理２３が施されている。ウェル１９同士の間のマイクロウェルプレート１７の表面が親水性であると、隣り合うウェル１９の内部の検体１５同士が、マイクロウェルプレート１７の表面を濡らす検体１５によってつながり、混ざってしまう恐れがある。したがって、ウェル１９の内面以外の部位において、マイクロウェルプレート１７の表面に疎水化処理２３を施すことで、マイクロウェルプレート１７の表面が検体１５によって濡れにくくなり、隣り合うウェル１９内の検体同士をより確実に分離することができる。 Further, the surface of the microwell plate 17 other than the well 19 is subjected to a hydrophobic treatment 23. If the surface of the microwell plate 17 between the wells 19 is hydrophilic, the specimens 15 in the adjacent wells 19 may be connected and mixed by the specimen 15 that wets the surface of the microwell plate 17. . Therefore, by subjecting the surface of the microwell plate 17 to the hydrophobic treatment 23 at a portion other than the inner surface of the well 19, the surface of the microwell plate 17 is less likely to be wetted by the specimen 15, and the specimens in the adjacent wells 19 are separated from each other. Separation can be performed more reliably.
一方、このように、マイクロウェルプレート１７の表面に疎水化処理２３を施すことで、マイクロウェルプレート１７上に滴下された検体１５は、玉状になる。したがって、検体１５は、マイクロウェルプレート１７の表面の一部にのみ留まり、マイクロウェルプレート１７の全体に広がることがない。 On the other hand, by subjecting the surface of the microwell plate 17 to the hydrophobic treatment 23 in this way, the specimen 15 dropped on the microwell plate 17 becomes a ball. Therefore, the specimen 15 stays only on a part of the surface of the microwell plate 17 and does not spread over the entire microwell plate 17.
次に、検体導入部材１を検体１５の近傍に配置する。図４（ａ）は、検体１５の近傍に検体導入部材１を配置した状態を示す側面図、図４（ｂ）は正面図である。検体導入部材１は、例えば、自動化された装置で柄部７が保持されて動作する。検体導入部材１は、ヘラ部３の検体接触面５が検体１５と対向するように配置される。この際、ヘラ部３の下端は、マイクロウェルプレート１７と接触する。 Next, the sample introduction member 1 is disposed in the vicinity of the sample 15. 4A is a side view showing a state in which the sample introduction member 1 is disposed in the vicinity of the sample 15, and FIG. 4B is a front view. The sample introduction member 1 operates by holding the handle 7 with an automated apparatus, for example. The sample introduction member 1 is arranged so that the sample contact surface 5 of the spatula part 3 faces the sample 15. At this time, the lower end of the spatula portion 3 comes into contact with the microwell plate 17.
図５（ａ）は、この状態から、検体導入部材１を検体１５の方向に移動させた状態を示す側面図、図５（ｂ）は正面図である。検体導入部材１を検体１５の方向に移動させると（図中矢印Ｄ）、検体導入部材１が検体１５へ接触する。前述した様に、ヘラ部３の検体接触面５の一部には、親水部９が形成されており、検体１５は、親水部９に対して濡れやすい。このため、検体導入部材１の検体接触面５の一部に接触した検体１５は、ヘラ部３に形成された親水部９の形成範囲に広げられる。すなわち、検体１５は、ヘラ部３の下端近傍の幅方向に広げられる。 FIG. 5A is a side view showing a state where the sample introduction member 1 is moved in the direction of the sample 15 from this state, and FIG. 5B is a front view. When the sample introduction member 1 is moved in the direction of the sample 15 (arrow D in the figure), the sample introduction member 1 contacts the sample 15. As described above, the hydrophilic part 9 is formed on a part of the specimen contact surface 5 of the spatula part 3, and the specimen 15 is easily wetted with the hydrophilic part 9. For this reason, the sample 15 that has contacted a part of the sample contact surface 5 of the sample introduction member 1 is expanded to the formation range of the hydrophilic portion 9 formed in the spatula portion 3. That is, the specimen 15 is expanded in the width direction near the lower end of the spatula portion 3.
この状態で、検体導入部材１（ヘラ部３）で、マイクロウェルプレート１７の表面をなぞることで、検体１５を、ウェル１９に導入することができる。この際、検体１５が、ヘラ部３の幅方向に広げられているため、マイクロウェルプレート１７の全面に対して検体１５を行きわたらせ、各ウェル１９へ効率よく検体１５を導入することができる。 In this state, the specimen 15 can be introduced into the well 19 by tracing the surface of the microwell plate 17 with the specimen introduction member 1 (spatial portion 3). At this time, since the specimen 15 is expanded in the width direction of the spatula portion 3, the specimen 15 can be spread over the entire surface of the microwell plate 17, and the specimen 15 can be efficiently introduced into each well 19.
以上、本実施の形態によれば、マイクロウェルプレート１７上で、玉状になりやすい検体１５を、容易にマイクロウェルプレート１７の全面に広げて、効率よく、検体１５を各ウェル１９に導入することができる。特に、ウェル１９の内面に親水化処理２１が施されているため、検体１５がウェル１９内に導入されやすく、ウェル１９内にエア溜りなどが生じにくい。 As described above, according to the present embodiment, on the microwell plate 17, the specimen 15 that tends to be ball-shaped is easily spread over the entire surface of the microwell plate 17, and the specimen 15 is efficiently introduced into each well 19. be able to. In particular, since the hydrophilic treatment 21 is performed on the inner surface of the well 19, the specimen 15 is easily introduced into the well 19, and air accumulation or the like is unlikely to occur in the well 19.
また、親水部９の上方には、疎水部１１が設けられる。このため、マイクロウェルプレート１７と接触することがない、ヘラ部３の上方に、検体１５が広がることを抑制することができる。このため、ヘラ部３の上部に付着する検体１５の量が削減され、検体１５を無駄なくウェル１９に導入することができる。特に、疎水部１１が親水部９の上方に１ｍｍ以上あれば、検体１５が親水部９の上方に付着することをより確実に抑制することができる。なお、親水部９の幅方向の中央部近傍は、液面の高さが高くなりやすいため、疎水部１１を親水部９の上方に２ｍｍ以上形成することで、より確実に、検体１５が親水部９の上方に付着することをより確実に抑制することができる。 A hydrophobic part 11 is provided above the hydrophilic part 9. For this reason, it can suppress that the sample 15 spreads above the spatula part 3 which does not contact the microwell plate 17. For this reason, the amount of the specimen 15 adhering to the upper portion of the spatula portion 3 is reduced, and the specimen 15 can be introduced into the well 19 without waste. In particular, if the hydrophobic part 11 is 1 mm or more above the hydrophilic part 9, it is possible to more reliably suppress the specimen 15 from adhering above the hydrophilic part 9. Since the height of the liquid surface tends to be high near the center of the hydrophilic portion 9 in the width direction, the specimen 15 is more reliably made hydrophilic by forming the hydrophobic portion 11 2 mm or more above the hydrophilic portion 9. It can suppress more reliably that it adheres above the part 9. FIG.
次に、第２の実施形態について説明する。なお、以下の説明において、検体導入部材１と同一の機能を奏する構成については、図１〜図５等と同一の符号を付し、重複する説明を省略する。図６は、第２の実施形態にかかる検体導入部材１ａの正面図である。 Next, a second embodiment will be described. In the following description, components having the same functions as those of the sample introduction member 1 are denoted by the same reference numerals as those in FIGS. 1 to 5 and the like, and redundant description is omitted. FIG. 6 is a front view of the sample introduction member 1a according to the second embodiment.
検体導入部材１ａは、検体導入部材１とほぼ同様の構成であるが、親水部９の形態が異なる。検体導入部材１ａは、疎水部１１が、親水部９の上方のみでなく、親水部９の周囲を囲むように形成される。すなわち、検体導入部材１ａでは、親水部９は、検体導入部材１ａ（ヘラ部３）の全幅に形成されず、疎水部１１が親水部９の幅方向の両側と上方とに形成される。なお、親水部９の両側の疎水部１１の幅は、それぞれ１ｍｍ以上であることが望ましい。 The sample introduction member 1a has substantially the same configuration as the sample introduction member 1, but the form of the hydrophilic portion 9 is different. The sample introduction member 1 a is formed so that the hydrophobic portion 11 surrounds the periphery of the hydrophilic portion 9 as well as the upper portion of the hydrophilic portion 9. In other words, in the sample introduction member 1a, the hydrophilic portion 9 is not formed in the entire width of the sample introduction member 1a (the spatula portion 3), and the hydrophobic portion 11 is formed on both sides in the width direction of the hydrophilic portion 9 and above. The width of the hydrophobic part 11 on both sides of the hydrophilic part 9 is preferably 1 mm or more.
次に、検体導入部材１ａを用いた、マイクロウェルへの検体の導入方法について説明する。図７（ａ）は、検体１５に検体導入部材１ａを接触させた状態を示す正面図、図７（ｂ）は平面図である。まず、前述した様に、マイクロウェルプレート１７上に検体１５を滴下し、検体導入部材１ａを検体１５の近傍に配置する。この状態から、検体導入部材１ａを検体１５の方向に移動させると、検体導入部材１ａが検体１５へ接触する。 Next, a method for introducing the sample into the microwell using the sample introduction member 1a will be described. FIG. 7A is a front view showing a state in which the sample introduction member 1a is brought into contact with the sample 15, and FIG. 7B is a plan view. First, as described above, the specimen 15 is dropped on the microwell plate 17 and the specimen introduction member 1a is disposed in the vicinity of the specimen 15. From this state, when the sample introduction member 1 a is moved in the direction of the sample 15, the sample introduction member 1 a comes into contact with the sample 15.
前述した様に、検体導入部材１ａの検体接触面５の一部に接触した検体１５は、ヘラ部３に形成された親水部９の形成範囲に広げられる。すなわち、検体１５は、ヘラ部３の下端近傍の幅方向に広げられる。 As described above, the specimen 15 that has contacted a part of the specimen contact surface 5 of the specimen introduction member 1 a is expanded to the formation range of the hydrophilic portion 9 formed in the spatula portion 3. That is, the specimen 15 is expanded in the width direction near the lower end of the spatula portion 3.
図７（ｂ）に示すように、この状態で、検体導入部材１ａ（ヘラ部３）で、マイクロウェルプレート１７の表面をなぞることで（図中矢印Ｈ）、検体１５を、ウェル１９に導入することができる。この際、検体１５は、親水部９の幅に広げられているため、親水部９の幅に対応する範囲Ｇの範囲に対して検体１５を行きわたらせ、各ウェル１９へ効率よく検体１５を導入することができる。 As shown in FIG. 7B, in this state, the specimen 15 is introduced into the well 19 by tracing the surface of the microwell plate 17 (arrow H in the figure) with the specimen introduction member 1a (the spatula portion 3). can do. At this time, since the specimen 15 is expanded to the width of the hydrophilic portion 9, the specimen 15 is spread over the range of the range G corresponding to the width of the hydrophilic portion 9, and the specimen 15 is efficiently introduced into each well 19. can do.
一方、図７（ｃ）は、検体１５に検体導入部材１を接触させて移動させた状態を示す平面図である。親水部９が全幅に形成された検体導入部材１の場合には、検体１５は、ヘラ部３の全幅に広げられる。このため、検体導入部材１を移動させると（図中矢印Ｉ）、検体１５の量によっては、検体１５がヘラ部３の両側方から後方にこぼれ落ちる（図中矢印Ｊ）。検体導入部材１の両側方からこぼれ落ちた検体１５は、マイクロウェルプレート１７上で再び玉状にまとまり、ウェル１９への導入効率が悪くなるおそれがある。 On the other hand, FIG. 7C is a plan view showing a state in which the sample introduction member 1 is moved in contact with the sample 15. In the case of the sample introduction member 1 in which the hydrophilic portion 9 is formed to the full width, the sample 15 is expanded to the full width of the spatula portion 3. For this reason, when the sample introduction member 1 is moved (arrow I in the figure), the sample 15 spills backward from both sides of the spatula portion 3 depending on the amount of the sample 15 (arrow J in the figure). The specimen 15 spilled from both sides of the specimen introduction member 1 is collected again in the shape of a ball on the microwell plate 17 and the introduction efficiency into the well 19 may be deteriorated.
これに対し、検体導入部材１ａは、検体１５がヘラ部３の幅方向に広がる範囲が規制されるため、検体１５をより確実に、マイクロウェルプレート１７の全面に行きわたらせて、効率よくウェル１９に検体１５を導入することができる。 On the other hand, the sample introduction member 1a restricts the range in which the sample 15 extends in the width direction of the spatula portion 3, so that the sample 15 is more reliably distributed over the entire surface of the microwell plate 17 and efficiently the well 19 The specimen 15 can be introduced into
第２の実施の形態によれば、第１の実施の形態と同様の効果を得ることができる。また、検体導入部材１ａは、親水部９がヘラ部３の幅方向の全幅に形成されず、親水部９の両側に疎水部１１が形成されるため、検体１５がヘラ部３の幅方向に広がる範囲が規制される。このため、検体導入部材１ａでマイクロウェルプレート１７をなぞる際に、検体１５がヘラ部３の後方にこぼれ落ちることを抑制することができる。 According to the second embodiment, an effect similar to that of the first embodiment can be obtained. Further, in the sample introduction member 1 a, the hydrophilic portion 9 is not formed in the full width in the width direction of the spatula portion 3, and the hydrophobic portions 11 are formed on both sides of the hydrophilic portion 9, so that the sample 15 extends in the width direction of the spatula portion 3. The range of expansion is regulated. For this reason, when the microwell plate 17 is traced by the sample introduction member 1a, the sample 15 can be prevented from spilling behind the spatula portion 3.
次に、第３の実施形態について説明する。図８は、第３の実施形態にかかる検体導入部材１ｂの分解斜視図、図９は、組立斜視図である。検体導入部材１ｂは、検体導入部材１等とほぼ同様の構成であるが、柄部に代えてディスポチップ１３が用いられる点で異なる。 Next, a third embodiment will be described. FIG. 8 is an exploded perspective view of the specimen introducing member 1b according to the third embodiment, and FIG. 9 is an assembled perspective view. The sample introduction member 1b has substantially the same configuration as the sample introduction member 1 and the like, but differs in that a disposable chip 13 is used instead of the handle.
検体導入部材１ｂのヘラ部３には、保持部２５が設けられる。保持部２５は、検体導入部材１ｂの検体接触面５（親水部９および疎水部１１が形成された面）の親水部９および疎水部１１の上方（ヘラ部３の上端近傍）に設けられる。保持部２５は略筒状である。保持部２５にはディスポチップ１３を装着可能であり、保持部２５によって、ディスポチップ１３の先端が保持される。 The spatula portion 3 of the sample introduction member 1b is provided with a holding portion 25. The holding part 25 is provided above the hydrophilic part 9 and the hydrophobic part 11 (near the upper end of the spatula part 3) on the specimen contact surface 5 (surface on which the hydrophilic part 9 and the hydrophobic part 11 are formed) of the specimen introduction member 1b. The holding part 25 is substantially cylindrical. The disposable chip 13 can be attached to the holding unit 25, and the tip of the disposable chip 13 is held by the holding unit 25.
保持部２５にディスポチップ１３を装着した状態において、ディスポチップ１３の先端は、親水部９の上方に位置する。すなわち、ディスポチップ１３の先端は、ヘラ部３の下端から所定の高さに位置する。 In a state where the disposable chip 13 is attached to the holding part 25, the tip of the disposable chip 13 is located above the hydrophilic part 9. That is, the tip of the disposable chip 13 is located at a predetermined height from the lower end of the spatula portion 3.
次に、検体導入部材１ｂを用いた、ウェルへの検体の導入方法について説明する。まず、ディスポチップ１３を、図示を省略した分注器に取り付け、検体貯留部（図示せず）に移動させ、検体をディスポチップ１３で吸引させる。次に、検体を保持した状態のディスポチップ１３を、ヘラ部３（保持部２５）に装着する。 Next, a method for introducing the specimen into the well using the specimen introduction member 1b will be described. First, the disposable chip 13 is attached to a dispenser (not shown), moved to a sample reservoir (not shown), and the sample is aspirated by the disposable chip 13. Next, the disposable chip 13 holding the sample is attached to the spatula portion 3 (holding portion 25).
次に、図１０（ａ）に示すように、検体導入部材１ｂを、あらかじめセットされたマイクロウェルプレート１７上に移動し、所定量の検体１５をマイクロウェルプレート１７上に滴下する（図中矢印Ｌ）。すなわち、ヘラ部３の検体接触面５の前方に、検体１５を滴下する。 Next, as shown in FIG. 10A, the sample introduction member 1b is moved onto the microwell plate 17 set in advance, and a predetermined amount of the sample 15 is dropped onto the microwell plate 17 (arrow in the figure). L). That is, the specimen 15 is dropped in front of the specimen contact surface 5 of the spatula portion 3.
次に、図１０（ｂ）に示すように、検体導入部材１ｂを検体１５の方向に移動させると（図中矢印Ｍ）、検体導入部材１ｂが検体１５へ接触する。前述した様に、ヘラ部３の検体接触面５の一部には、親水部９が形成されており、検体１５は、親水部９に対して濡れやすい。このため、検体導入部材１ｂの検体接触面５の一部に接触した検体１５は、ヘラ部３に形成された親水部９の形成範囲に広げられる。すなわち、検体１５は、ヘラ部３の下端近傍の幅方向に広げられる。 Next, as shown in FIG. 10B, when the sample introduction member 1b is moved in the direction of the sample 15 (arrow M in the figure), the sample introduction member 1b comes into contact with the sample 15. As described above, the hydrophilic part 9 is formed on a part of the specimen contact surface 5 of the spatula part 3, and the specimen 15 is easily wetted with the hydrophilic part 9. For this reason, the specimen 15 that has contacted a part of the specimen contact surface 5 of the specimen introduction member 1 b is expanded to the formation range of the hydrophilic portion 9 formed in the spatula portion 3. That is, the specimen 15 is expanded in the width direction near the lower end of the spatula portion 3.
この状態で、検体導入部材１（ヘラ部３）で、マイクロウェルプレート１７の表面をなぞることで、検体１５を、ウェルに導入することができる。この際、検体１５が、ヘラ部３の幅方向に広げられているため、効率よく、マイクロウェルプレート１７の全面に対して、検体１５を行きわたらせ、各ウェルへ効率よく検体１５を導入することができる。 In this state, the specimen 15 can be introduced into the well by tracing the surface of the microwell plate 17 with the specimen introduction member 1 (spatial portion 3). At this time, since the specimen 15 is expanded in the width direction of the spatula portion 3, the specimen 15 is efficiently distributed over the entire surface of the microwell plate 17, and the specimen 15 is efficiently introduced into each well. Can do.
第３の実施の形態によれば、第１の実施の形態と同様の効果を得ることができる。また、検体導入部材１ｂは、ヘラ部３にディスポチップ１３を装着可能であるため、検体１５の滴下と、ヘラ部３による検体１５のウェルへの導入を連続して行うことができる。このため、効率よく、検体１５を各ウェルに導入することができる。なお、ディスポチップ１３を保持可能な保持部２５は、親水部９等が形成されない従来のヘラ部にも適用可能である。 According to the third embodiment, an effect similar to that of the first embodiment can be obtained. In addition, since the sample introduction member 1b can be mounted with the disposable chip 13 on the spatula portion 3, the sample 15 can be dropped and introduced to the well of the sample 15 by the spatula portion 3 continuously. For this reason, the specimen 15 can be efficiently introduced into each well. The holding part 25 that can hold the disposable chip 13 can also be applied to a conventional spatula part in which the hydrophilic part 9 or the like is not formed.
以上、添付図を参照しながら、本発明の実施の形態を説明したが、本発明の技術的範囲は、前述した実施の形態に左右されない。当業者であれば、特許請求の範囲に記載された技術的思想の範疇内において各種の変更例または修正例に想到し得ることは明らかであり、それらについても当然に本発明の技術的範囲に属するものと了解される。 As mentioned above, although embodiment of this invention was described referring an accompanying drawing, the technical scope of this invention is not influenced by embodiment mentioned above. It is obvious for those skilled in the art that various modifications or modifications can be conceived within the scope of the technical idea described in the claims, and these are naturally within the technical scope of the present invention. It is understood that it belongs.
１、１ａ、１ｂ………検体導入部材
３………ヘラ部
５………検体接触面
７………柄部
９………親水部
１１………疎水部
１３………ディスポチップ
１５………検体
１７………マイクロウェルプレート
１９………ウェル
２１………親水化処理
２３………疎水化処理
２５………保持部
1, 1a, 1b... Specimen introduction member 3... Spatula part 5... Specimen contact surface 7 ...... Pattern part 9 ...... Hydrophilic part 11 ...... Hydrophobic part 13. …… Sample 17 ………… Microwell plate 19 ………… Well 21 ………… Hydrophilic treatment 23 ………… Hydrophobic treatment 25 ……… Holding part
Claims (5)
前記導入部材は板状であり、前記導入部材の少なくとも一方の面が、前記検体との接触面であって、前記検体との接触面の下端部近傍であって前記マイクロウェルプレートとの接触部近傍の少なくとも一部に親水部が形成され、少なくとも前記親水部の上方に疎水部が形成されることを特徴とする検体導入部材。 An introduction member for introducing a specimen into a well formed in a microwell plate,
The introduction member is plate-shaped, and at least one surface of the introduction member is a contact surface with the sample, and is in the vicinity of a lower end portion of the contact surface with the sample and a contact portion with the microwell plate at least part of the vicinity of the hydrophilic portion is formed on the analyte introduction member, wherein a hydrophobic portion is formed above at least the hydrophilic portion.
前記導入部材は板状であり、前記導入部材の少なくとも一方の面が、前記検体との接触面であって、前記検体との接触面の下端部近傍であって前記マイクロウェルプレートとの接触部近傍の少なくとも一部に親水部が形成され、
前記親水部は、前記検体導入部材の全幅に形成されず、少なくとも前記親水部の幅方向の両側に疎水部が形成されることを特徴とする検体導入部材。 An introduction member for introducing a specimen into a well formed in a microwell plate,
The introduction member is plate-shaped, and at least one surface of the introduction member is a contact surface with the sample, and is in the vicinity of a lower end portion of the contact surface with the sample and a contact portion with the microwell plate A hydrophilic part is formed in at least a part of the vicinity,
The sample introduction member, wherein the hydrophilic portion is not formed over the entire width of the sample introduction member, and a hydrophobic portion is formed at least on both sides in the width direction of the hydrophilic portion.
前記検体導入部材は板状であり、前記検体導入部材の少なくとも一方の面が、検体との接触面であって、前記検体との接触面の下端部近傍であって前記マイクロウェルプレートとの接触部近傍の表面の一部に親水部が形成され、少なくとも前記親水部の上方に疎水部が形成されており、
複数のウェルが形成された前記マイクロウェルプレート上に、ディスポチップで検体を滴下し、
前記検体に前記検体導入部材を接触させて、前記検体を前記親水部の形成範囲に広げ、前記検体導入部材で前記マイクロウェルプレートの表面をなぞり、前記検体を、前記ウェルに導入することを特徴とするウェルへの検体導入方法。 A method for introducing a sample into a well formed in a microwell plate using a sample introduction member,
The sample introduction member is plate-shaped, and at least one surface of the sample introduction member is a contact surface with the sample, and is in the vicinity of the lower end portion of the contact surface with the sample and in contact with the microwell plate. hydrophilic portion is formed on part of the surface of the parts near and the hydrophobic part is formed above at least the hydrophilic portion,
A specimen is dropped with a disposable chip on the microwell plate on which a plurality of wells are formed,
The sample introduction member is brought into contact with the sample, the sample is spread over the formation range of the hydrophilic portion, the surface of the microwell plate is traced with the sample introduction member, and the sample is introduced into the well. Sample introduction method to the well.
前記保持部にディスポチップを装着し、
前記マイクロウェルプレート上に、前記ディスポチップで検体を滴下した後、前記検体導入部材で前記マイクロウェルプレートの表面をなぞり、前記検体を、前記ウェルに導入することを特徴とする請求項４記載のウェルへの検体導入方法。
On the surface where the hydrophilic part and the hydrophobic part of the sample introduction member are formed, a holding part for holding the tip of the disposable chip is provided above the hydrophilic part,
A disposable chip is attached to the holding part,
The microwell plate, wherein after dropping the sample in a disposable chip, the at sample introduction member tracing the surface of the microwell plate, the specimen, according to claim 4, wherein the introduction into the well Sample introduction method to well.
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