JPS62288547A - Apparatus for analyzing living body-related substance, bacterium and cell - Google Patents
Apparatus for analyzing living body-related substance, bacterium and cellInfo
- Publication number
- JPS62288547A JPS62288547A JP13148286A JP13148286A JPS62288547A JP S62288547 A JPS62288547 A JP S62288547A JP 13148286 A JP13148286 A JP 13148286A JP 13148286 A JP13148286 A JP 13148286A JP S62288547 A JPS62288547 A JP S62288547A
- Authority
- JP
- Japan
- Prior art keywords
- living body
- cell
- cells
- analyzer
- quartz vibrator
- 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.)
- Pending
Links
- 239000000126 substance Substances 0.000 title claims abstract description 26
- 241000894006 Bacteria Species 0.000 title abstract 4
- 239000010453 quartz Substances 0.000 claims abstract description 15
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000013078 crystal Substances 0.000 claims description 34
- 244000005700 microbiome Species 0.000 claims description 7
- 238000004458 analytical method Methods 0.000 claims description 4
- 230000001413 cellular effect Effects 0.000 claims 1
- 230000000813 microbial effect Effects 0.000 claims 1
- 238000005259 measurement Methods 0.000 abstract description 8
- 230000010355 oscillation Effects 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 230000032683 aging Effects 0.000 abstract description 2
- 230000003100 immobilizing effect Effects 0.000 abstract 2
- 210000004027 cell Anatomy 0.000 description 19
- 108090001090 Lectins Proteins 0.000 description 8
- 102000004856 Lectins Human genes 0.000 description 8
- 239000002523 lectin Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 210000003743 erythrocyte Anatomy 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 2
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 2
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 241000219871 Ulex Species 0.000 description 2
- 235000010730 Ulex europaeus Nutrition 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229950006780 n-acetylglucosamine Drugs 0.000 description 2
- JAPYIBBSTJFDAK-UHFFFAOYSA-N 2,4,6-tri(propan-2-yl)benzenesulfonyl chloride Chemical compound CC(C)C1=CC(C(C)C)=C(S(Cl)(=O)=O)C(C(C)C)=C1 JAPYIBBSTJFDAK-UHFFFAOYSA-N 0.000 description 1
- KYARBIJYVGJZLB-UHFFFAOYSA-N 7-amino-4-hydroxy-2-naphthalenesulfonic acid Chemical compound OC1=CC(S(O)(=O)=O)=CC2=CC(N)=CC=C21 KYARBIJYVGJZLB-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 241001539176 Hime Species 0.000 description 1
- SHZGCJCMOBCMKK-PQMKYFCFSA-N L-Fucose Natural products C[C@H]1O[C@H](O)[C@@H](O)[C@@H](O)[C@@H]1O SHZGCJCMOBCMKK-PQMKYFCFSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 101710197072 Lectin 1 Proteins 0.000 description 1
- 101710198419 Lectin 11 Proteins 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- SHZGCJCMOBCMKK-SXUWKVJYSA-N alpha-L-fucose Chemical compound C[C@@H]1O[C@@H](O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-SXUWKVJYSA-N 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 108010084962 lima bean lectin Proteins 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 229910052763 palladium Inorganic materials 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Abstract
Description
【発明の詳細な説明】
３、発明の詳細な説明
〔産業上の利用分野〕
本発明は、生体関連物質、微生物および細胞の分析装置
に関する。Detailed Description of the Invention 3. Detailed Description of the Invention [Field of Industrial Application] The present invention relates to a device for analyzing biological substances, microorganisms, and cells.
それぞれ異なる物質を固定化または化学的処理をした複
数の水晶振動子バイオセンサーを有する生体関連物質等
の測定装置によって、複数の生体関連物質の測定を可能
にした。また、何も固定化されていない水晶振動子を参
照用のセンサーとして用いた生体関連物質の測定装置に
よって非定常状態での発振周波数から参照用水晶振動子
との周波数を求めることによって迅速な測定が可能とな
った。It has become possible to measure a plurality of biologically related substances by using a measuring device for measuring biologically related substances, etc., which has a plurality of quartz crystal biosensors, each of which has a different substance immobilized or chemically treated. In addition, by using a bio-related substance measuring device that uses an unfixed crystal oscillator as a reference sensor, quick measurements can be made by determining the frequency with the reference crystal oscillator from the oscillation frequency in an unsteady state. became possible.
生体関連物質等の測定方法としては、これまで主に液体
クロマトグラフィーや電気泳動法、凝集を直接肉眼で確
認する方法などが用いられていた。Up until now, liquid chromatography, electrophoresis, and direct visual confirmation of aggregation have been used as methods for measuring biological substances.
これらの方法は、測定悪疫が低い、測定法が煩雑であっ
たり、測定に時間がかかるという問題点があった。These methods have problems in that the measurement rate is low, the measurement method is complicated, and the measurement takes time.
本発明では、高感度な生体物質の分析が可能である水晶
振動子バイオセンサーを複数使用し、生体関連物質また
は微生物、細胞の分析装置によって、さらに高感度な複
数の生体関連成分や微生物・細胞の分析が可能となった
。また、表面になにも固定化等の処理を行っていない水
晶振動子を参照用センサーとして用いた分析装置によっ
て発振周波数の短期的なエージングや溶液の導電率や温
度の影響による周波数のずれを取り除いて測定すること
ができるようになり、迅速な測定が可能となった。In the present invention, a plurality of quartz crystal biosensors capable of highly sensitive analysis of biological substances are used, and an analyzer for biological substances, microorganisms, and cells is used to analyze biological substances, microorganisms, and cells with even higher sensitivity. analysis became possible. In addition, using an analyzer that uses a quartz crystal resonator whose surface has not been subjected to any fixation or other treatment as a reference sensor, we can detect short-term aging of the oscillation frequency and frequency shifts due to the effects of the conductivity and temperature of the solution. It is now possible to remove and measure, making rapid measurements possible.
本発明の分析装置により、高感度な複数生体関連物質や
微生物・細胞の分析が連続的に迅速に行えるようになっ
た。With the analyzer of the present invention, highly sensitive analyzes of multiple biological substances, microorganisms, and cells can be performed continuously and quickly.
（ＡＢＯ式血液型センサー）
以下、図面に従い本発明のＡＢＯ式血液型センサーへの
応用について詳述する。(ABO blood type sensor) Hereinafter, the application of the present invention to an ABO blood type sensor will be described in detail with reference to the drawings.
第１図（ａｌは、水晶振動子バイオセンサーとセルの斜
視図である。水晶振動子１はＡＴカット９Ｍ１１ｚのも
のを用いた。各水晶振動子１の両面には電極２が設けら
れており、それぞれの電極２にはり−ド３が接続されて
いる。前記水晶振動子１を覆う様にセル４が設けられ、
該セルには該セル中に液をｉｌ過させる為にパイプ５が
取り付けられている。前記リード３は該セル４の外部へ
引き出されている。ま吹、第１図（ｂｌは、水晶振動子
１としてＧＴカット４ Ｍ ＩＩＺのものを用いた水晶
振動子バイオセンサーとセルの斜視図である。この４つ
の水晶振動子のうち３つの振動子にそれぞれ、リママメ
レクチン、ハンディラマメレクチン１１ハリエニシダレ
クチン■を固定化した。FIG. 1 (Al is a perspective view of a crystal oscillator biosensor and a cell. The crystal oscillator 1 used is an AT cut 9M11z. Electrodes 2 are provided on both sides of each crystal oscillator 1. , a beam 3 is connected to each electrode 2.A cell 4 is provided so as to cover the crystal resonator 1,
A pipe 5 is attached to the cell to allow liquid to pass through the cell. The lead 3 is led out of the cell 4. Figure 1 (bl is a perspective view of a crystal oscillator biosensor and cell using a GT cut 4M IIZ crystal oscillator 1. Three of these four crystal oscillators Lima bean lectin, handicraft lance lectin 11, and gorse lectin ■ were immobilized on the plates, respectively.
第１図（Ｃ１は、水晶振動子を用いた生体関連物質測定
装面の概略を示したものである。水晶振動子Ｉの表面上
に設けられた電極２からのびたり−ド３は、発振回路６
に接続されており、また該発振回路６は、周波数カウン
タ７に接続されている。Fig. 1 (C1 shows the outline of a biologically related substance measuring device using a crystal resonator.A electrode 3 extending from an electrode 2 provided on the surface of the crystal resonator I is used for oscillation. circuit 6
The oscillation circuit 6 is connected to a frequency counter 7.
前記水晶振動子１の外部を取り凹むセル４に設けられた
パイプ５の一方は該セル４中に液を通過される為のポン
プ９が接続されている。パイプ５の他の一端は電磁バル
ブ１０の出口に接続されている。電磁弁の入口は、もう
１つの電磁弁１１とサンプル注入口１２に接続されてい
る。電磁弁１１の入口は、ｐＨ７，５，０，０５ＭＩＪ
ン酸１夏衝ン夜だめ１３とｐ Ｈ７，５，０，１Ｍα−
Ｌ−フコース、０．１ＭＮ−アセチル−Ｄ−グルコサミ
ン？容？（ｌｚだめ１４に接続されている。前記周波数
カウンタ７、定量ポンプ９および＠磁弁１０，１１はコ
ンピュータに接続され、測定および制御が行われた。A pump 9 for passing liquid into the cell 4 is connected to one side of a pipe 5 provided in a cell 4 that takes in the outside of the crystal resonator 1. The other end of the pipe 5 is connected to the outlet of the electromagnetic valve 10. The inlet of the solenoid valve is connected to another solenoid valve 11 and to a sample inlet 12 . The inlet of the solenoid valve 11 has a pH of 7.5, 0.05 MIJ.
pH 7,5,0,1Mα-
L-fucose, 0.1M N-acetyl-D-glucosamine? Yong? (The frequency counter 7, metering pump 9, and magnetic valves 10 and 11 were connected to a computer for measurement and control.
水晶振動子バイオセンサーの作成は以下の手法に従った
。まず、水晶振動子の電極表面をパラジウムメッキし、
０．５Ｎ ＮａＯＨ中で１時間陽極酸化した。The quartz crystal biosensor was created according to the following method. First, the electrode surface of the crystal resonator is plated with palladium,
Anodized in 0.5N NaOH for 1 hour.
次に、５％トリシルクロライド、１．２％ピリジンを含
むアセトン溶液で３０分間処理を行い、５ｍＭＨｃｊ！
で洗浄した。レクチンの固定化は、それぞれｌ ｍｇ／
ｍ ｌに溶解させたレクチン溶液に水晶振動子を１時間
浸漬することによって行った。さらに０．１Ｍグリシン
溶液で３０分間反応させ未反応店の処理を行った。Next, treatment was performed for 30 minutes with an acetone solution containing 5% trisyl chloride and 1.2% pyridine, and 5mM Hcj!
Washed with. The immobilization of lectin was carried out at 1 mg/L, respectively.
The test was carried out by immersing the crystal resonator in a lectin solution dissolved in ml for 1 hour. Furthermore, the reaction was carried out for 30 minutes with a 0.1M glycine solution to treat unreacted portions.
本装置によるＡＢＯ式血液型の判定は、以下のようにし
て行った。ABO blood type determination using this device was performed as follows.
まず、３つのレクチンを固定化した水晶振動子と参照用
の水晶振動子は、あらかじめ、媛ｉＪｊ液を通液したセ
ルの中で発振させておき、電磁弁ＩＯを切り換え、サン
プル注入口１２から血球試料を導入した。電磁弁１０の
開閉により、試料は、如意の割合で希釈して測定を行っ
た。試料がセルを通過する際は、反応が十分進行するよ
うに、定量ポンプ９をＩＩ？′ＪｒＩして、流速が低く
なるようコントロールした。次に、リン酸緩衝液を流し
、非特異的な吸着を除去し、各水晶振動子の発振周波数
を測定し、３つのレクチンを固定化した水晶振動子の参
照用水晶振動子の差を求め、この値が、試料との反応前
とで変化しているかどうかによって、血液型の判定を行
った。First, the crystal oscillator on which the three lectins are immobilized and the reference crystal oscillator are oscillated in advance in a cell through which the Hime iJj solution has been passed, and the solenoid valve IO is switched to allow the sample injection port 12 to be oscillated. A blood cell sample was introduced. By opening and closing the solenoid valve 10, the sample was diluted at a desired ratio and measured. When the sample passes through the cell, the metering pump 9 is set to II to ensure that the reaction progresses sufficiently. 'JrI to control the flow rate to be low. Next, a phosphate buffer solution was poured to remove non-specific adsorption, the oscillation frequency of each crystal unit was measured, and the difference between the reference crystal units and the three lectin-immobilized crystal units was determined. The blood type was determined based on whether this value changed before the reaction with the sample.
この結果、リママメレクチンを固定化した水晶振動子は
、Ａ型因子を持つ赤血球だけに応答することがわかった
。パンディラマメレクチンＩを固定化した水晶振動子は
、Ａ型因子、Ｂ型因子ともに応答を示したが、Ｂ型因子
を持つ赤血球に対する応答が大きく、リママメレクチン
固定化水晶振動子の応答との比較によって、Ａ、Ｂ、Ａ
Ｂ型の判定ができることがわかった。ハリエニシダレク
チン■を固定化した水晶振動子は、０型の赤血球だけに
応答を示すことがわかった。As a result, it was found that the quartz crystal oscillator immobilized with limamame lectin responded only to red blood cells containing type A factor. The quartz crystal oscillator immobilized with limanium lectin I showed a response to both type A and type B factors, but the response to red blood cells containing type B factor was large, and the response was different from that of the quartz oscillator immobilized with limanium lectin. By comparing A, B, A
It was found that type B can be determined. It was found that a quartz crystal oscillator immobilized with gorse lectin ■ responded only to type 0 red blood cells.
次に、電磁弁１１を切り換え、ｐ Ｈ７，５，０，１Ｍ
α−Ｌ−フコース、０．１ＭＮ−アセチル−Ｄ−グルコ
サミン溶液をセル中に導入することによって、表面のレ
クチンに結合した赤血球を除去した。引き続き、ｐＨ７
，５’Ｊン酸緩衝液を十分通液した後、再び、測定を行
った。Next, switch the solenoid valve 11 and set the pH to 7, 5, 0, 1M.
Red blood cells bound to lectin on the surface were removed by introducing a solution of α-L-fucose and 0.1M N-acetyl-D-glucosamine into the cell. Continue to pH 7
, 5'J acid buffer was sufficiently passed through the tube, and the measurement was carried out again.
以上のように、本発明の分析装置により、ＡＢＯ式血液
型の測定が、連続的に迅速に行えるようになった。また
、ＧＴカット水晶振動子を用いた装置では、セルを小さ
くすることができるため、試料の量が少量ですむという
利点がある。As described above, the analyzer of the present invention enables continuous and rapid measurement of ABO blood types. Further, in an apparatus using a GT-cut crystal resonator, the cell can be made small, so there is an advantage that a small amount of sample is required.
本発明の生体関連物質の測定装置によって、高感度な複
数の生体関連物質、微生物、細胞の分析が連続的に迅速
に行えるようになった。With the measuring device for biologically related substances of the present invention, highly sensitive analyzes of multiple biologically related substances, microorganisms, and cells can be performed continuously and rapidly.
第１図（ａｌは、本発明の生体関連物質測定装置におけ
るＡＴカフト水晶振動子を用いたバイオセンサーとセル
の斜視図であり、第１図（ｂｌは、ＧＴカット水晶振動
子を用いた場合のバイオセンサーとセルの斜視図である
。第１図（Ｃ１は、本発明の測定装置を示すブロック図
である。
以上
出願人 セイコー電子工業株式会社
バイオセンナとセルの＠現図
第１ 図（ｂ）Figure 1 (al is a perspective view of a biosensor and cell using an AT-cut crystal oscillator in the biologically related substance measuring device of the present invention, and Figure 1 (bl is a perspective view of a cell using a GT-cut crystal oscillator) Fig. 1 (C1 is a block diagram showing the measuring device of the present invention. b)
Claims (3)
って、生体関連物質の濃度や成分を分析する装置におい
て、複数の水晶振動子を有する生体関連物質、微生物お
よび細胞の分析装置。(1) An analyzer for analyzing biological substances, microorganisms, and cells having a plurality of crystal oscillators, which analyzes the concentration and components of biological substances by changing the frequency due to changes in the weight of the surface of a crystal oscillator.
、化学的な処理をした水晶振動子をセンサーとして用い
たことを特徴とする特許請求の範囲第１項記載の生体関
連物質、微生物および細胞の分析装置。(2) Biological substances, microorganisms, and cells according to claim 1, characterized in that a crystal oscillator having different types of substances immobilized on its surface or chemically treated is used as a sensor. analysis equipment.
固定化された物質を持たない水晶振動子を用いることを
特徴とする特許請求の範囲第１項記載の生体関連物質、
微生物および細胞の分析装置。(3) A biologically related substance according to claim 1, characterized in that at least one of the plurality of crystal oscillators uses a quartz crystal oscillator that does not have a substance immobilized on its surface;
Microbial and cellular analysis equipment.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP13148286A JPS62288547A (en) | 1986-06-06 | 1986-06-06 | Apparatus for analyzing living body-related substance, bacterium and cell |
| EP86307115A EP0215669A3 (en) | 1985-09-17 | 1986-09-16 | Analytical device and method for analysis of biochemicals, microbes and cells |
| US06/908,371 US4789804A (en) | 1985-09-17 | 1986-09-17 | Analytical device and method utilizing a piezoelectric crystal biosensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP13148286A JPS62288547A (en) | 1986-06-06 | 1986-06-06 | Apparatus for analyzing living body-related substance, bacterium and cell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS62288547A true JPS62288547A (en) | 1987-12-15 |
Family
ID=15059009
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP13148286A Pending JPS62288547A (en) | 1985-09-17 | 1986-06-06 | Apparatus for analyzing living body-related substance, bacterium and cell |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62288547A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02226043A (en) * | 1989-02-27 | 1990-09-07 | Seiko Instr Inc | Reaction measuring apparatus |
| JPH04307351A (en) * | 1991-04-04 | 1992-10-29 | Hitachi Ltd | Method for detecting trace of gaseous component in atmosphere and applied machinery |
| JPH04337462A (en) * | 1991-05-13 | 1992-11-25 | Inax Corp | Quantitative analyzing method for total protein in urine |
| JPH05322884A (en) * | 1992-05-22 | 1993-12-07 | Inax Corp | Method for measuring bio-component |
| JPH0694591A (en) * | 1992-09-14 | 1994-04-05 | Sogo Yatsukou Kk | Detection method for biochemical substance and biochemical substance detection sensor used for the method |
| JP2008512673A (en) * | 2004-09-16 | 2008-04-24 | コリア インスティテュート オブ サイエンス アンド テクノロジー | Method and system for detecting biological components |
-
1986
- 1986-06-06 JP JP13148286A patent/JPS62288547A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02226043A (en) * | 1989-02-27 | 1990-09-07 | Seiko Instr Inc | Reaction measuring apparatus |
| JPH04307351A (en) * | 1991-04-04 | 1992-10-29 | Hitachi Ltd | Method for detecting trace of gaseous component in atmosphere and applied machinery |
| JPH04337462A (en) * | 1991-05-13 | 1992-11-25 | Inax Corp | Quantitative analyzing method for total protein in urine |
| JPH05322884A (en) * | 1992-05-22 | 1993-12-07 | Inax Corp | Method for measuring bio-component |
| JP2673980B2 (en) * | 1992-05-22 | 1997-11-05 | 株式会社イナックス | Measuring method of biological components |
| JPH0694591A (en) * | 1992-09-14 | 1994-04-05 | Sogo Yatsukou Kk | Detection method for biochemical substance and biochemical substance detection sensor used for the method |
| JP2008512673A (en) * | 2004-09-16 | 2008-04-24 | コリア インスティテュート オブ サイエンス アンド テクノロジー | Method and system for detecting biological components |
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