JPH04330272A - Incubation apparatus - Google Patents
Incubation apparatusInfo
- Publication number
- JPH04330272A JPH04330272A JP12526691A JP12526691A JPH04330272A JP H04330272 A JPH04330272 A JP H04330272A JP 12526691 A JP12526691 A JP 12526691A JP 12526691 A JP12526691 A JP 12526691A JP H04330272 A JPH04330272 A JP H04330272A
- Authority
- JP
- Japan
- Prior art keywords
- reaction
- reaction vessel
- heating
- cooling
- reaction container
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000011534 incubation Methods 0.000 title claims description 22
- 238000006243 chemical reaction Methods 0.000 claims abstract description 100
- 238000010438 heat treatment Methods 0.000 claims abstract description 27
- 238000001816 cooling Methods 0.000 claims abstract description 19
- 239000000463 material Substances 0.000 claims description 13
- 229910052751 metal Inorganic materials 0.000 claims description 11
- 239000002184 metal Substances 0.000 claims description 11
- 229920003002 synthetic resin Polymers 0.000 claims description 10
- 239000000057 synthetic resin Substances 0.000 claims description 10
- 239000002826 coolant Substances 0.000 claims description 9
- 230000004544 DNA amplification Effects 0.000 abstract description 3
- 238000000137 annealing Methods 0.000 abstract description 2
- 230000015572 biosynthetic process Effects 0.000 abstract description 2
- 230000000694 effects Effects 0.000 abstract description 2
- 230000036425 denaturation Effects 0.000 abstract 1
- 238000004925 denaturation Methods 0.000 abstract 1
- 238000004904 shortening Methods 0.000 abstract 1
- 238000003786 synthesis reaction Methods 0.000 abstract 1
- 229920002545 silicone oil Polymers 0.000 description 20
- 239000000523 sample Substances 0.000 description 7
- 239000010409 thin film Substances 0.000 description 7
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 239000012488 sample solution Substances 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 239000010408 film Substances 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 2
- 239000004809 Teflon Substances 0.000 description 2
- 229920006362 Teflon® Polymers 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000002480 mineral oil Substances 0.000 description 2
- 235000010446 mineral oil Nutrition 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 108091028026 C-DNA Proteins 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
Landscapes
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Sustainable Development (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Thermal Sciences (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Devices For Use In Laboratory Experiments (AREA)
Abstract
Description
【０００１】0001
【産業上の利用分野】本発明は、例えば遺伝子増幅反応
（ＰＣＲ反応）等に利用することのできるインキュベー
ション装置に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an incubation device that can be used, for example, in gene amplification reactions (PCR reactions).
【０００２】0002
【従来の技術】例えばＰＣＲ反応を行なうには、サンプ
ル溶液を充填した反応容器を、異なった温度で順次加熱
冷却するサイクルを複数回繰り返す必要がある。従来、
そのようなＰＣＲ反応に用いるインキュベーション装置
は、反応容器を加熱冷却する単一の加熱冷却用ブロック
を備えたものであった。そして、そのブロックをヒータ
ー又はクーラーにより所定の温度まで加熱冷却すること
により、そのブロックに挿入される反応容器を加熱冷却
し、次にその加熱冷却用ブロックを異なった所定の温度
にまで温度調節するという動作を繰り返すことによりＰ
ＣＲ反応を行なっていた。2. Description of the Related Art For example, in order to carry out a PCR reaction, it is necessary to repeat a cycle of successively heating and cooling a reaction vessel filled with a sample solution several times at different temperatures. Conventionally,
The incubation device used for such a PCR reaction was equipped with a single heating and cooling block that heated and cooled the reaction container. Then, by heating and cooling the block to a predetermined temperature using a heater or cooler, the reaction vessel inserted into the block is heated and cooled, and then the temperature of the heating and cooling block is adjusted to a different predetermined temperature. By repeating this action, P
A CR reaction was being carried out.
【０００３】0003
【発明が解決しようとする課題】従来のインキュベーシ
ョン装置にあっては、反応容器を加熱冷却するためのブ
ロックが単一であったため、反応容器を異なった温度に
調節するには、そのたびに加熱冷却用ブロックの温度制
御を行なう必要があって無駄な時間、電力が必要となっ
ていた。[Problems to be Solved by the Invention] Conventional incubation devices have a single block for heating and cooling the reaction container, so in order to adjust the reaction container to a different temperature, it is necessary to heat the reaction container each time. It was necessary to control the temperature of the cooling block, which required wasted time and power.
【０００４】また、反応容器の材質としては反応試薬と
反応しないことと作成の容易さから合成樹脂材等が用い
られているが、熱伝導率が悪いためにサンプル溶液を目
的温度に加熱冷却するのに無駄な時間や電力が必要以上
にかかるという問題があった。[0004] In addition, synthetic resin materials are used as the material for the reaction container because they do not react with the reaction reagent and are easy to manufacture, but because of their poor thermal conductivity, the sample solution must be heated and cooled to the target temperature. However, there was a problem in that it required more time and energy than necessary.
【０００５】本発明は上記従来技術の問題を解決するこ
とを目的とする。The present invention aims to solve the problems of the prior art described above.
【０００６】[0006]
【課題を解決するための手段】本発明の特徴とするとこ
ろは、予め互いに異なった温度に温度調節された複数の
反応容器加熱冷却用媒体を備え、各加熱冷却用媒体は互
いに独立して一定の順序で前記反応容器に接触される点
にある。その反応容器が金属薄膜を合成樹脂により被覆
した材料から形成されているが好ましい。[Means for Solving the Problems] The present invention is characterized in that it is equipped with a plurality of reaction vessel heating and cooling media whose temperatures are adjusted to different temperatures in advance, and each heating and cooling medium is maintained at a constant temperature independently of each other. The points are contacted with the reaction vessel in this order. Preferably, the reaction vessel is made of a material in which a thin metal film is coated with a synthetic resin.
【０００７】[0007]
【作用】本発明の構成によるインキュベーション装置に
より反応容器を加熱冷却するには、まず各加熱冷却用媒
体を予め互いに異なった温度に調節する。具体的な温度
は反応の種類に応じて定める。次に、各加熱冷却用媒体
により反応容器を順次加熱冷却する。これにより、各加
熱冷却用媒体による加熱冷却順序に従って反応容器は異
なった温度に調節される。各加熱冷却用媒体による反応
容器の加熱冷却順序は反応の種類に応じて一定のものと
する。[Operation] In order to heat and cool a reaction container using the incubation apparatus constructed according to the present invention, first, each heating and cooling medium is adjusted to different temperatures in advance. The specific temperature is determined depending on the type of reaction. Next, the reaction vessel is sequentially heated and cooled using each heating and cooling medium. Thereby, the temperature of the reaction vessel is adjusted to different temperatures according to the order of heating and cooling by each heating and cooling medium. The order of heating and cooling the reaction vessel using each heating and cooling medium is constant depending on the type of reaction.
【０００８】反応容器が金属薄膜を合成樹脂により被覆
した材料から形成されることにより、反応容器は熱伝導
率がよく反応試薬と反応しないものとなる。[0008] Since the reaction container is formed from a material in which a thin metal film is coated with a synthetic resin, the reaction container has good thermal conductivity and does not react with the reaction reagent.
【０００９】[0009]
【実施例】以下、図面を参照して本発明の実施例を説明
する。Embodiments Hereinafter, embodiments of the present invention will be described with reference to the drawings.
【００１０】図１および図２に示すインキュベーション
装置１は複数の加熱冷却用媒体を備えている。本実施例
では、加熱冷却用媒体は３つのブロック２ａ、２ｂ、２
ｃにより構成されている。各ブロック２ａ、２ｂ、２ｃ
は図１において左右方向に沿って並列し、内部にヒータ
ーもしくは冷却素子（図示省略）を内蔵し、そのヒータ
ーもしくは冷却素子により各ブロック２ａ、２ｂ、２ｃ
は互いに異なった温度に調節される。また、各ブロック
２ａ、２ｂ、２ｃにそれぞれ複数の上向開口の凹部３ａ
、３ｂ、３ｃが形成されている。それら凹部３ａ、３ｂ
、３ｃは図２において左右方向に沿って並列する。The incubation apparatus 1 shown in FIGS. 1 and 2 is equipped with a plurality of heating and cooling media. In this embodiment, the heating and cooling medium is used in three blocks 2a, 2b, 2
It is composed of c. Each block 2a, 2b, 2c
are arranged in parallel along the left and right direction in FIG.
are adjusted to different temperatures. In addition, each block 2a, 2b, 2c has a plurality of upwardly opening recesses 3a.
, 3b, 3c are formed. Those recesses 3a, 3b
, 3c are arranged in parallel along the left-right direction in FIG.
【００１１】図３に示すように、ブロック２ａ、２ｂ、
２ｃが上方に移動することで凹部３ａ、３ｂ、３ｃに反
応容器４が挿入され、これにより反応容器４の加熱冷却
が行なわれるものとされている。また、各ブロック２ａ
、２ｂ、２ｃは互いに独立して一定の順序で反応容器４
を加熱冷却可能とされている。As shown in FIG. 3, blocks 2a, 2b,
The reaction container 4 is inserted into the recesses 3a, 3b, and 3c by moving the container 2c upward, thereby heating and cooling the reaction container 4. In addition, each block 2a
, 2b, 2c are arranged in a certain order independently of each other in the reaction vessel 4.
can be heated and cooled.
【００１２】すなわち、各ブロック２ａ、２ｂ、２ｃは
それぞれ台車１４にガイドレール５を介して上下動可能
に取り付けられると共に、それぞれ流体圧シリンダ６ａ
、６ｂ、６ｃにより上下駆動される。その台車１４は、
流体圧シリンダ７により図１においてレール１５上で左
右方向に移動可能とされる。前記反応容器４を図２にお
いて左方から右方に搬送するベルトコンベヤ８が設けら
れ、このベルトコンベヤ８により反応容器４は、ブロッ
ク２ａ、２ｂ、２ｃの上方において保持される。すなわ
ち、図４に示すようにベルトコンベヤ８のベルト８ａに
は、ブロック２ａ、２ｂ、２ｃの凹部３ａ、３ｂ、３ｃ
の形成ピッチと同ピッチで保持孔９が形成され、反応容
器４にはフランジ４ａが形成されている。これにより、
保持孔９に反応容器４を挿入すると共に、フランジ４ａ
をベルト８ａの上面に接するものとすることで、反応容
器４はベルトコンベヤ８に保持される。That is, each of the blocks 2a, 2b, 2c is attached to a truck 14 via a guide rail 5 so as to be movable up and down, and each block is connected to a hydraulic cylinder 6a.
, 6b, and 6c. The trolley 14 is
The hydraulic cylinder 7 allows movement in the left-right direction on the rail 15 in FIG. A belt conveyor 8 is provided to convey the reaction container 4 from left to right in FIG. 2, and the belt conveyor 8 holds the reaction container 4 above the blocks 2a, 2b, and 2c. That is, as shown in FIG. 4, the belt 8a of the belt conveyor 8 has recesses 3a, 3b, 3c of blocks 2a, 2b, 2c
Holding holes 9 are formed at the same pitch as the formation pitch, and a flange 4a is formed in the reaction vessel 4. This results in
While inserting the reaction container 4 into the holding hole 9, the flange 4a
The reaction container 4 is held by the belt conveyor 8 by making it in contact with the upper surface of the belt 8a.
【００１３】上記インキュベーション装置１を用いてＰ
ＣＲ反応を行なう場合の動作を説明する。なお、ＰＣＲ
反応における温度サイクルは、図６に示すように、反応
容器に充填されたサンプルをまず９４℃の温度に１分間
保つことで熱変性させ、次に５５℃の温度に１分間保つ
ことでプライマーのアニーリングを行ない、次に７２℃
の温度に１分間保つことでＤＮＡ鎖の合成を行なう。こ
の温度サイクルを１６回繰り返すものとする。この場合
、各ブロック２ａ、２ｂ、２ｃに形成される凹部３ａ、
３ｂ、３ｃの数は、温度サイクルの繰り返し回数と等し
く１６とされる。[0013] Using the above incubation device 1, P
The operation when performing the CR reaction will be explained. In addition, PCR
As shown in Figure 6, the temperature cycle in the reaction is such that the sample filled in the reaction vessel is first kept at a temperature of 94°C for 1 minute to denature it, and then kept at a temperature of 55°C for 1 minute to denature the primer. Annealing and then 72℃
DNA strands are synthesized by keeping the temperature at 1 min for 1 minute. This temperature cycle shall be repeated 16 times. In this case, recesses 3a formed in each block 2a, 2b, 2c,
The numbers 3b and 3c are 16, which is equal to the number of repetitions of the temperature cycle.
【００１４】まず、各ブロック２ａ、２ｂ、２ｃを予め
９４℃、５４℃、７２℃にそれぞれ温度調節する。また
、流体圧シリンダ７により台車４を移動させることで、
９４℃に温度調節したブロック２ａをベルトコンベヤ８
のベルト８ａの下方に位置させる。また、ベルトコンベ
ヤ８の反応容器４の搬送方向後端に位置する凹部３ａの
上方位置で、サンプル溶液を充填した反応容器４をベル
トコンベヤ８により保持する。First, the temperatures of the blocks 2a, 2b, and 2c are adjusted in advance to 94°C, 54°C, and 72°C, respectively. In addition, by moving the cart 4 using the fluid pressure cylinder 7,
The block 2a whose temperature has been adjusted to 94°C is transferred to the belt conveyor 8.
is located below the belt 8a. Further, the belt conveyor 8 holds the reaction container 4 filled with the sample solution at a position above the recess 3 a located at the rear end of the reaction container 4 in the conveyance direction of the belt conveyor 8 .
【００１５】この状態から、図５に示すように、まず９
４℃に温度調節されたブロック２ａを流体圧シリンダ６
ａにより上方移動させ、凹部３ａ内に反応容器４を挿入
して加熱し、これによりサンプルの熱変性を行なう。次
に、ブロック２ａを下降させ、台車４を移動させること
で、５５℃に温度調節したブロック２ｂを反応容器４の
下方に位置させる。しかる後に、その５５℃に温度調節
したブロック２ｂを上方移動させ、凹部３ｂ内に反応容
器４を挿入して冷却して一定温度に保ち、これにより、
サンプルのプライマーのアニーリングを行なう。次に、
ブロック２ｂを下降させ、台車４を移動させることで、
反応容器４の下方に７２℃に温度調節したブロック２ｃ
を位置させる。しかる後に、その７２℃に温度調節した
ブロック２ｃを上方移動させ、凹部３ｃ内に反応容器４
を挿入して加熱して一定温度に保ち、これにより、サン
プルのＤＮＡ鎖の合成を行なう。次に、ブロック２ｃを
下降させ、台車４を移動させることで、反応容器４の下
方に９４℃に温度調節されたブロック２ａを位置させる
。また、ベルトコンベヤ８を凹部３ａ、３ｂ、３ｃの１
ピッチ分だけ搬送方向前方に移動させ、搬送方向後端に
位置する凹部３ａの上方位置で、サンプル溶液を充填し
た新たな反応容器４をベルトコンベヤ８により保持する
。しかる後に、各ブロック２ａ、２ｂ、２ｃによる反応
容器４の加熱冷却を上記と同様に行なう。From this state, as shown in FIG.
The block 2a whose temperature is adjusted to 4°C is transferred to the fluid pressure cylinder 6.
a, and the reaction container 4 is inserted into the recess 3a and heated, thereby thermally denaturing the sample. Next, by lowering the block 2a and moving the cart 4, the block 2b whose temperature has been adjusted to 55° C. is positioned below the reaction vessel 4. After that, the block 2b whose temperature has been adjusted to 55°C is moved upward, and the reaction vessel 4 is inserted into the recess 3b to cool it and maintain it at a constant temperature.
Anneal the sample primers. next,
By lowering the block 2b and moving the trolley 4,
A block 2c whose temperature is controlled to 72°C is placed below the reaction vessel 4.
position. Thereafter, the block 2c whose temperature has been adjusted to 72°C is moved upward, and the reaction vessel 4 is placed in the recess 3c.
is inserted and heated to maintain a constant temperature, thereby synthesizing the DNA strand of the sample. Next, by lowering the block 2c and moving the cart 4, the block 2a whose temperature is adjusted to 94° C. is positioned below the reaction container 4. Also, the belt conveyor 8 is connected to one of the recesses 3a, 3b, and 3c.
The belt conveyor 8 holds a new reaction container 4 filled with the sample solution by moving it forward in the conveyance direction by the pitch, and at a position above the recess 3a located at the rear end in the conveyance direction. Thereafter, the reaction vessel 4 is heated and cooled by the blocks 2a, 2b, and 2c in the same manner as described above.
【００１６】以上の動作を１６回繰り返すことで、最初
に反応が行なわれた反応容器４の内部のサンプルの処理
が終了し、この反応容器４をベルトコンベヤ８から取り
外す。この取外しは作業者が手で行なってもよいし、あ
るいはマニピュレータを用いて行なってもよい。By repeating the above operation 16 times, processing of the sample inside the reaction container 4 in which the reaction was first performed is completed, and this reaction container 4 is removed from the belt conveyor 8. This removal may be performed manually by an operator, or may be performed using a manipulator.
【００１７】上記実施例によれば、互いに異なった温度
に調節されるブロック２ａ、２ｂ、２ｃを用い、しかも
、バッチ処理でなく逐次処理によりＰＣＲ反応を行なう
ことができ、無駄な時間、電力を従来よりも大幅に減少
させることができる。According to the above embodiment, the blocks 2a, 2b, and 2c which are adjusted to different temperatures can be used, and the PCR reaction can be carried out by sequential processing instead of batch processing, thereby saving wasted time and power. This can be significantly reduced compared to conventional methods.
【００１８】なお、インキュベーション装置１を例えば
シーケンスコントローラーにより作動制御し、その制御
プログラムを変更することで、異なった温度サイクルの
反応にも対応できるようにするのが好ましい。It is preferable to control the operation of the incubation apparatus 1 using, for example, a sequence controller, and to change the control program so as to be able to respond to reactions with different temperature cycles.
【００１９】また、処理中に反応溶液が蒸発しないよう
に、反応溶液の上部にミネラルオイルを載せたり、反応
容器４に蓋を溶着するようにしてもよい。また、ブロッ
ク２ａ、２ｂ、２ｃの凹部３ａ、３ｂ、３ｃの内部にミ
ネラルオイルを充填することで、反応溶液とブロック２
ａ、２ｂ、２ｃとの間の熱伝導を向上するようにしても
よい。Furthermore, in order to prevent the reaction solution from evaporating during processing, mineral oil may be placed on top of the reaction solution, or a lid may be welded to the reaction container 4. Also, by filling the insides of the recesses 3a, 3b, 3c of the blocks 2a, 2b, 2c with mineral oil, the reaction solution and the blocks 2
A, 2b, and 2c may be improved in heat conduction.
【００２０】図７〜図１１を参照して本発明の第２実施
例を説明する。A second embodiment of the present invention will be described with reference to FIGS. 7 to 11.
【００２１】図７に示すインキュベーション装置５１は
、反応容器５２を保持する保持具５３と、３基のタンク
５４ａ、５４ｂ、５４ｃを備えている。各タンク５４ａ
、５４ｂ、５４ｃには、反応容器５２の加熱冷却用媒体
としてシリコンオイルが貯留される。また、各タンク５
４ａ、５４ｂ、５４ｃの加熱用ヒーターもしくは冷却素
子（図示省略）が設けられ、各タンク５４ａ、５４ｂ、
５４ｃに貯留されたシリコンオイルは互いに異なった温
度に調節される。各タンク５４ａ、５４ｂ、５４ｃから
シリコンオイルを導出する導出管５５ａ、５５ｂ、５５
ｃの途中に、それぞれ開閉弁５６ａ、５６ｂ、５６ｃが
設けられている。各導出管５５ａ、５５ｂ、５５ｃは互
いに連結されて単一の供給管５７とされている。この供
給管５７は前記保持具５３に接続されている。The incubation device 51 shown in FIG. 7 includes a holder 53 for holding a reaction container 52, and three tanks 54a, 54b, and 54c. Each tank 54a
, 54b, and 54c, silicone oil is stored as a medium for heating and cooling the reaction vessel 52. In addition, each tank 5
4a, 54b, 54c heating heaters or cooling elements (not shown) are provided, and each tank 54a, 54b,
The silicone oils stored in 54c are adjusted to different temperatures. Outlet pipes 55a, 55b, 55 that lead out silicone oil from each tank 54a, 54b, 54c
On-off valves 56a, 56b, and 56c are provided in the middle of c, respectively. The respective outlet pipes 55a, 55b, and 55c are connected to each other to form a single supply pipe 57. This supply pipe 57 is connected to the holder 53.
【００２２】この保持具５３は上向開口のシリコンオイ
ル貯留層とされ、供給管５７から供給されるシリコンオ
イルが貯留される。この貯留されたオイルを排出するた
めに保持具５３に形成された排出孔５３ａにパイプ５８
が接続され、このパイプ５８の途中にポンプ５９が設け
られている。そのパイプ５８は分岐されて前記タンク５
４ａ、５４ｂ、５４ｃに接続される。このパイプ５８か
ら各タンク５４ａ、５４ｂ、５４ｃへの分岐管５８ａ、
５８ｂ、５８ｃの途中に、それぞれ開閉弁５９ａ、５９
ｂ、５９ｃが設けられている。なお、保持具５３の内部
に貯留されたシリコンオイルの温度制御用のヒーターお
よびクーラーを設けるのが好ましい。The holder 53 is a silicone oil storage layer with an upward opening, and stores silicone oil supplied from the supply pipe 57. A pipe 58 is connected to a discharge hole 53a formed in the holder 53 to discharge this stored oil.
is connected, and a pump 59 is provided in the middle of this pipe 58. The pipe 58 is branched to the tank 5.
4a, 54b, and 54c. A branch pipe 58a from this pipe 58 to each tank 54a, 54b, 54c,
On-off valves 59a and 59 are provided in the middle of 58b and 58c, respectively.
b, 59c are provided. Note that it is preferable to provide a heater and a cooler for temperature control of the silicone oil stored inside the holder 53.
【００２３】反応容器５２は、図９および図１０に示す
ように、金属薄膜６０を樹脂６１により被覆した材料か
ら形成されており、反応溶液を入れる多数の上向開口の
凹部６２を有する。金属薄膜６０としては、例えばアル
ミニウムのような熱伝導率の良いものを用いる。また、
金属薄膜６０を被覆する合成樹脂６１としては、例えば
テフロンのような反応試薬と反応しないものを用いる。
その凹部６２の数は本実施例では９６個とされる。この
反応容器５２により保持具５３の上向の開口を覆うよう
にすることで、反応容器５２は保持具５３により保持さ
れる。なお、保持具５３の内部に、反応容器５２を支持
するための支柱５３ｂが互いに間隔をおいて複数設けら
れている。As shown in FIGS. 9 and 10, the reaction container 52 is made of a material in which a metal thin film 60 is coated with a resin 61, and has a number of upwardly opening recesses 62 into which the reaction solution is placed. As the metal thin film 60, a material having good thermal conductivity such as aluminum is used, for example. Also,
As the synthetic resin 61 covering the metal thin film 60, a material that does not react with the reaction reagent, such as Teflon, is used. The number of recesses 62 is 96 in this embodiment. The reaction container 52 is held by the holder 53 by covering the upward opening of the holder 53 with the reaction container 52 . Note that a plurality of pillars 53b for supporting the reaction container 52 are provided inside the holder 53 at intervals.
【００２４】図８に示すように、反応容器５２は、アル
ミニウム等の金属薄膜６０をテフロン等の合成樹脂６１
により被覆した材料から成る蓋６３により覆われる。そ
の蓋６３と反応容器５２とは接着材等で接着するように
する。また、蓋６３を反応容器５２に押し付けると共に
保温するため、その蓋６３を、内部にヒーターを内蔵し
たプレート６４と前記保持具５３とにより挟み込む。As shown in FIG. 8, the reaction vessel 52 includes a metal thin film 60 made of aluminum or the like and a synthetic resin 61 made of Teflon or the like.
It is covered with a lid 63 made of a material coated with. The lid 63 and the reaction container 52 are bonded together using an adhesive or the like. Further, in order to press the lid 63 against the reaction container 52 and keep it warm, the lid 63 is sandwiched between the holder 53 and a plate 64 having a built-in heater therein.
【００２５】上記インキュベーション装置５１を用いて
ＰＣＲ反応を行なう場合の動作を説明する。なお、ＰＣ
Ｒ反応の温度サイクルは前記図６に示した条件と同様と
する。The operation when performing a PCR reaction using the incubation device 51 will be explained. In addition, PC
The temperature cycle of the R reaction is the same as the conditions shown in FIG. 6 above.
【００２６】まず、反応容器５２にサンプル溶液を充填
し、蓋６３により密閉し、しかる後に保持具５３とプレ
ート６４により反応容器５２を挟み込むようにして保持
する。また、タンク５４ａ、５４ｂ、５４ｃに貯留され
たシリコンオイルを予め９４℃、５４℃、７２℃にそれ
ぞれ温度調節する。First, the reaction container 52 is filled with a sample solution and sealed with a lid 63, and then the reaction container 52 is held between the holder 53 and the plate 64 so as to be sandwiched therebetween. Further, the temperature of the silicone oil stored in the tanks 54a, 54b, and 54c is adjusted in advance to 94°C, 54°C, and 72°C, respectively.
【００２７】次に、開閉弁５６ａを開くことにより、タ
ンク５４ａに貯留された９４℃に温度調節されたシリコ
ンオイルを供給管５７を介して保持具５３の内部に供給
する。この供給は、保持具５３に設けられた開口５３ｃ
からシリコンオイルがオーバーフローするまで行ない、
供給後に開閉弁５６ａを閉じる。この際、保持具５３を
ヒーターにより９４℃に温度調節するのが好ましい。こ
れにより、９４℃に温度調節されたシリコンオイルによ
り反応容器５２を加熱し、サンプルの熱変性を行なう。Next, by opening the on-off valve 56a, silicone oil whose temperature has been adjusted to 94° C. stored in the tank 54a is supplied to the inside of the holder 53 via the supply pipe 57. This supply is carried out through an opening 53c provided in the holder 53.
Continue until the silicone oil overflows.
After supplying, the on-off valve 56a is closed. At this time, it is preferable to adjust the temperature of the holder 53 to 94° C. using a heater. Thereby, the reaction vessel 52 is heated with silicone oil whose temperature is controlled to 94° C., and the sample is thermally denatured.
【００２８】次に、開閉弁５９ａを開くと共にポンプ５
９を駆動することで、保持具５３内のシリコンオイルを
タンク５４ａに還流し、還流後に開閉弁５９ａを閉じる
と共にポンプ５９を停止する。Next, the on-off valve 59a is opened and the pump 5 is turned on.
By driving 9, the silicone oil in the holder 53 is returned to the tank 54a, and after the return, the on-off valve 59a is closed and the pump 59 is stopped.
【００２９】次に、開閉弁５６ｂを開き、供給管５７を
介して保持具５３の内部に５５℃に温度調節されたシリ
コンオイルを供給する。この供給は開口５３ｃからシリ
コンオイルがオーバーフローするまで行ない、供給後に
開閉弁５６ｂを閉じる。この際、保持具５３をクーラー
により５５℃まで冷却するのが好ましい。これにより、
反応容器５２を５５℃のシリコンオイルで冷却し、サン
プルのプライマーのアニーリングを行なう。Next, the on-off valve 56b is opened and silicone oil whose temperature is adjusted to 55° C. is supplied into the holder 53 through the supply pipe 57. This supply is continued until the silicone oil overflows from the opening 53c, and after the supply, the on-off valve 56b is closed. At this time, it is preferable to cool the holder 53 to 55° C. using a cooler. This results in
The reaction vessel 52 is cooled with silicone oil at 55° C., and the sample primer is annealed.
【００３０】次に、開閉弁５９ｂを開くと共にポンプ５
９を駆動し、保持具５３内のシリコンオイルをタンク５
４ｂに還流し、還流後に開閉弁５９ａを閉じると共にポ
ンプ５９を停止する。Next, open the on-off valve 59b and turn off the pump 5.
9 to drain the silicone oil in the holder 53 into the tank 5.
4b, and after the reflux, the on-off valve 59a is closed and the pump 59 is stopped.
【００３１】次に、開閉弁５６ｃを開き、供給管を介し
て保持具５３の内部に７２℃に温度調節されたシリコン
オイルを供給する。この供給は開口５３ｃからシリコン
オイルがオーバーフローするまで行ない、供給後に開閉
弁５６ｃを閉じる。この際、保持具５３をヒーターによ
り７２℃に温度調節するのが好ましい。これにより、シ
リコンオイルにより反応容器５２を加熱し、ＤＮＡ鎖の
合成を行なう。しかる後に、開閉弁５９ｃを開くと共に
ポンプ５９を駆動することでシリコンオイルをタンク５
４ｃに還流し、還流後に開閉弁５９ｃを閉じると共にポ
ンプ５９を停止する。Next, the on-off valve 56c is opened and silicone oil whose temperature is adjusted to 72° C. is supplied into the holder 53 through the supply pipe. This supply is continued until the silicone oil overflows from the opening 53c, and after the supply, the on-off valve 56c is closed. At this time, it is preferable to adjust the temperature of the holder 53 to 72° C. using a heater. Thereby, the reaction vessel 52 is heated by the silicone oil, and DNA chains are synthesized. After that, by opening the on-off valve 59c and driving the pump 59, the silicone oil is pumped into the tank 5.
4c, and after the reflux, the on-off valve 59c is closed and the pump 59 is stopped.
【００３２】以上により、一回の温度サイクルを完了す
る。この温度サイクルを１６回行なうことで反応を終了
する。With the above steps, one temperature cycle is completed. The reaction is completed by performing this temperature cycle 16 times.
【００３３】上記第２実施例によれば、互いに異なった
温度に調節されるシリコンオイルにより反応容器５２を
加熱冷却することができるので、無駄な時間を従来より
も低減することができる。しかも、反応容器５２は金属
薄膜６０を合成樹脂により被覆した材料から形成されて
いるので、熱伝導率がよく、かつ反応試薬と反応するこ
とがないものとできる。また、容易に製造することがで
きるので、通常は使い捨てられる反応容器として好まし
いものである。According to the second embodiment, since the reaction vessel 52 can be heated and cooled using silicone oils that are adjusted to different temperatures, wasted time can be reduced compared to the conventional method. Moreover, since the reaction container 52 is made of a material in which the metal thin film 60 is coated with a synthetic resin, it has good thermal conductivity and does not react with the reaction reagent. Moreover, since it can be easily manufactured, it is preferable as a reaction container that is normally disposable.
【００３４】なお、図１１に示すように金属薄膜を合成
樹脂により被覆した帯状材料をリール６７に巻取り、こ
のリールから繰り出す材料を金型６５、６６を用いて反
応容器５２に成形する成形装置を、インキュベーション
装置に組み込むようにしてもよい。As shown in FIG. 11, there is a molding device that winds up a strip-shaped material in which a metal thin film is coated with a synthetic resin onto a reel 67, and molds the material fed from the reel into the reaction vessel 52 using molds 65 and 66. may be incorporated into the incubation device.
【００３５】[0035]
【発明の効果】本発明によるインキュベーション装置に
よれば、互いに異なった温度に調節される加熱冷却用媒
体を用いて反応容器を一定の順序で加熱冷却することが
でき、反応容器に充填されたサンプルの処理時間を短縮
でき、かつ電力を最小限にできる。また、金属薄膜を合
成樹脂により被覆した材料から反応容器を形成すること
で、反応容器を熱伝導率がよく、かつ反応試薬と反応し
ないものとすることができ、処理時間および電力の無駄
を低減することができる。Effects of the Invention According to the incubation device of the present invention, the reaction vessels can be heated and cooled in a fixed order using heating and cooling media that are adjusted to different temperatures, and the samples filled in the reaction vessels can be heated and cooled in a certain order. processing time can be shortened and power consumption can be minimized. In addition, by forming the reaction container from a material in which a thin metal film is coated with a synthetic resin, the reaction container has good thermal conductivity and does not react with the reaction reagent, reducing waste of processing time and power. can do.
【図１】本発明の第１実施例に係るインキュベーション
装置の正面図FIG. 1 is a front view of an incubation device according to a first embodiment of the present invention.
【図２】本発明の第１実施例に係るインキュベーション
装置の側面図FIG. 2 is a side view of the incubation device according to the first embodiment of the present invention.
【図３】本発明の第１実施例に係る加熱冷却用媒体の側
面図FIG. 3 is a side view of the heating and cooling medium according to the first embodiment of the present invention.
【図４】本発明の第１実施例に係るインキュベーション
装置の部分平面図FIG. 4 is a partial plan view of the incubation device according to the first embodiment of the present invention.
【図５】本発明の第１実施例に係るインキュベーション
装置の作用説明図FIG. 5 is an explanatory diagram of the operation of the incubation device according to the first embodiment of the present invention.
【図６】ＰＣＲ反応の温度条件を示す図[Figure 6] Diagram showing temperature conditions for PCR reaction
【図７】本発明
の第２実施例に係るインキュベーション装置の構成説明
図FIG. 7 is an explanatory diagram of the configuration of an incubation device according to a second embodiment of the present invention.
【図８】本発明の第２実施例に係るインキュベーション
装置の部分断面図FIG. 8 is a partial sectional view of an incubation device according to a second embodiment of the present invention.
【図９】本発明の第２実施例に係る反応容器の断面図FIG. 9 is a sectional view of a reaction vessel according to a second embodiment of the present invention.
【
図１０】本発明の第２実施例に係る反応容器の平面図[
FIG. 10: Plan view of a reaction container according to the second embodiment of the present invention
【
図１１】本発明の第２実施例に係る反応容器の製造装置
の構成説明図[
FIG. 11 is an explanatory diagram of the configuration of a reaction container manufacturing apparatus according to a second embodiment of the present invention
１ インキュベーション装置
２ａ、２ｂ、２ｃ ブロック（加熱冷却用媒体）４
反応容器
５１ インキュベーション装置
５２ 反応容器
６０ 金属薄膜
６１ 合成樹脂1 Incubation devices 2a, 2b, 2c blocks (heating and cooling medium) 4
Reaction container 51 Incubation device 52 Reaction container 60 Metal thin film 61 Synthetic resin
Claims (2)
れた複数の反応容器加熱冷却用媒体を備え、各加熱冷却
用媒体は互いに独立して一定の順序で前記反応容器に接
触されることを特徴とするインキュベーション装置。1. A plurality of reaction vessel heating/cooling media whose temperatures are adjusted to different temperatures in advance are provided, and each heating/cooling medium is brought into contact with the reaction vessel in a fixed order independently of each other. Incubation device.
被覆した材料から形成されていることを特徴とする請求
項１に記載のインキュベーション装置。2. The incubation device according to claim 1, wherein the reaction container is made of a material in which a thin metal film is coated with a synthetic resin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP12526691A JP3070134B2 (en) | 1991-04-26 | 1991-04-26 | Incubation device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP12526691A JP3070134B2 (en) | 1991-04-26 | 1991-04-26 | Incubation device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH04330272A true JPH04330272A (en) | 1992-11-18 |
| JP3070134B2 JP3070134B2 (en) | 2000-07-24 |
Family
ID=14905828
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP12526691A Expired - Lifetime JP3070134B2 (en) | 1991-04-26 | 1991-04-26 | Incubation device |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP3070134B2 (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5504007A (en) * | 1989-05-19 | 1996-04-02 | Becton, Dickinson And Company | Rapid thermal cycle apparatus |
| US5576218A (en) * | 1994-01-11 | 1996-11-19 | Abbott Laboratories | Method for thermal cycling nucleic acid assays |
| US5960976A (en) * | 1996-10-18 | 1999-10-05 | Ngk Insulators, Ltd. | Sample container and method for producing the same |
| JP2001136954A (en) * | 1999-08-31 | 2001-05-22 | Toshiba Corp | Device for treating nucleic acid and method for treating nucleic acid |
| WO2003006162A3 (en) * | 2001-07-13 | 2003-05-22 | Univ British Columbia | Thermal cycling methods and apparatus |
| JP2011516882A (en) * | 2008-04-10 | 2011-05-26 | デナトール アクティエボラグ | Device for storing and preparing biological samples |
| EP2714276A1 (en) * | 2011-05-24 | 2014-04-09 | Ingeny PCR B.V. | System for and method of changing temperatures of substances |
| JP2016112015A (en) * | 2014-12-16 | 2016-06-23 | ヤマトエスロン株式会社 | Dna amplification method, dna amplification structure, and dna detector using the same |
| WO2020004655A1 (en) * | 2018-06-28 | 2020-01-02 | 株式会社ツーセル | Biological sample warming method, biological sample warming vessel, and kit for warming biological sample |
-
1991
- 1991-04-26 JP JP12526691A patent/JP3070134B2/en not_active Expired - Lifetime
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5504007A (en) * | 1989-05-19 | 1996-04-02 | Becton, Dickinson And Company | Rapid thermal cycle apparatus |
| US5576218A (en) * | 1994-01-11 | 1996-11-19 | Abbott Laboratories | Method for thermal cycling nucleic acid assays |
| US5960976A (en) * | 1996-10-18 | 1999-10-05 | Ngk Insulators, Ltd. | Sample container and method for producing the same |
| US6140613A (en) * | 1996-10-18 | 2000-10-31 | Ngk Insulators, Ltd | PCR method for amplifying a gene using metallic sample container having inner surface coated with a resin or metal oxide |
| JP2001136954A (en) * | 1999-08-31 | 2001-05-22 | Toshiba Corp | Device for treating nucleic acid and method for treating nucleic acid |
| WO2003006162A3 (en) * | 2001-07-13 | 2003-05-22 | Univ British Columbia | Thermal cycling methods and apparatus |
| JP2011516882A (en) * | 2008-04-10 | 2011-05-26 | デナトール アクティエボラグ | Device for storing and preparing biological samples |
| EP2714276A1 (en) * | 2011-05-24 | 2014-04-09 | Ingeny PCR B.V. | System for and method of changing temperatures of substances |
| US10076757B2 (en) | 2011-05-24 | 2018-09-18 | Molecular Biology Systems B.V. | System for and method of changing temperatures of substances |
| EP2714276B1 (en) * | 2011-05-24 | 2024-10-09 | Molecular Biology Systems B.V. | System for and method of changing temperatures of substances |
| JP2016112015A (en) * | 2014-12-16 | 2016-06-23 | ヤマトエスロン株式会社 | Dna amplification method, dna amplification structure, and dna detector using the same |
| WO2020004655A1 (en) * | 2018-06-28 | 2020-01-02 | 株式会社ツーセル | Biological sample warming method, biological sample warming vessel, and kit for warming biological sample |
Also Published As
| Publication number | Publication date |
|---|---|
| JP3070134B2 (en) | 2000-07-24 |
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