公开(公告)号：JPS63194723A

公开(公告)日：1988-08-11

申请号：JP2583587

申请日：1987-02-05

Applicant: NEC CORP

Inventor： KAWANA YOSHIE ,  KIMURA JUN

IPC: G01N1/00 ,  B01F11/00 ,  B01F11/02 ,  G01N1/36

Abstract: PURPOSE:To miniaturize the device and to measure a chemical substance of very small amount with high accuracy, by constituting the agitating device for very small amount of solution with a vibrator inserted into a sample solution and a piezoelectric actuator vibrating the vibrator. CONSTITUTION:The piezoelectric actuator 4 consist of an element the shape of which is changed according to the impressed voltage. When the vibrator 2 is brought in contact with the piezoelectric actuator 4 impressed with an optional pulse voltage, the optional vibration is given to the vibrator. When a top end of the vibrator 2 is inserted into the sample solution 3, the solution is efficiently agitated by the vibration of the vibrator. The agitation is carried out in every case where the solution is contained in a microvessel, where a flow cell is used in a closed flow, or where a solution drop is put on a hydrophobic base plate.

公开(公告)号：JPS63111454A

公开(公告)日：1988-05-16

申请号：JP25892186

申请日：1986-10-29

Applicant: NEC CORP

Inventor： KAWANA YOSHIE ,  KIMURA JUN

IPC: G01N27/414 ,  G01N27/30 ,  G01N27/327

Abstract: PURPOSE:To exactly form a pattern of an enzyme-contg. soln. so that a crosslinking treatment can be made in that form by dropping the enzyme-contg. soln. to gate parts on a solid surface, then bringing the same into contact with vapor contg. a crosslinking reagent to insolubilize the soln. CONSTITUTION:Three pieces of ion sensitive field effect type transistors are formed of silicon formed in an island shape on a sapphire substrate 2. Ureaze immobilized films 12, glucose oxidase immobilized films 13 and albumin films 14 are formed on the respective gate parts 1 by dropping of the solns. by an ink jet method. After the films are once dried, the substrate is placed on a supporting net in a hermetic vessel contg. an aq. glutaraldehyde soln. of 25% and is rested for about 5min to insolubilize the enzyme films.

公开(公告)号：JPS61234349A

公开(公告)日：1986-10-18

申请号：JP7575885

申请日：1985-04-10

Applicant: NEC CORP

Inventor： KAWANA YOSHIE ,  KIMURA JUN ,  KURIYAMA TOSHIHIDE

IPC: G01N27/00 ,  C12Q1/00 ,  G01N27/30 ,  G01N27/414

Abstract: PURPOSE:To form a biosensor having several kinds of enzyme immobilized films, by a method wherein an enzyme immobilized film and a photoresist layer are formed on a substrate provided with an ion sensor and a process of removing all but a portion required therefor. CONSTITUTION:An ion induction section 2 of an FE type ion sensor is provided on the top surface of a substrate 1 and an enzyme immobilized film 3 and a photoresist layer 4a are formed over the entire surface thereof. Then, the photoresist layer 4a is removed except for a specified surface of the ion induction section 2a by exposure and development and the enzyme immobilized film 3 is decomposed. Thereafter, a second enzyme immobilized film 5 and a photoresist layer 4b are formed on the top surface of the ion induction section and the same removing process is repeated to form a biosensor having different enzyme immobilized films 3 and 5 on the ion induction section 2. Thus, a massproducible fine multi-biosensor can be obtained by applying an IC production technology.

公开(公告)号：JPH01207657A

公开(公告)日：1989-08-21

申请号：JP3181688

申请日：1988-02-16

Applicant: NEC CORP

Inventor： KAWANA YOSHIE ,  KIMURA JUN

IPC: G01N27/416 ,  G01N37/00

Abstract: PURPOSE:To improve measurement accuracy by measuring the concns. of 1st and 2nd chemical materials by using 1st and 2nd diluting liquids and determining the concns. of the chemical materials in a sample soln. by using the 1st and 2nd measured concn. values. CONSTITUTION:A glucose sensor chip 1 is immersed into, for example, 20mul diluting liquid and 10mul standard solns. of various concns. are exactly added from an injection port 3 thereto. A calibration curve is obtd. by the concns. of the standard solns. of this time as well as an ISFET amplifier 4 and a signal processor 5. The similar calibration curve when the volume of the diluting liquid is changed to 40mul is obtd. The chip 1 is immersed into 20mul diluting liquid by using the calibration curves obtd. in such a manner and the potential of this time is designated as 0. A proper volume of the sample soln. having about 150mg/dl glucose concn. is added from the injection port 3 to the liquid and a change in output voltage is determined. Further, 20mul diluting liquid (2nd diluting liquid) is added thereto and the output voltage is measured. The measurement is made by changing the volume of the sample soln. The fairly exact concns. are obtainable even if the volume of the sample soln. varies by about 20%.

公开(公告)号：JPS61133853A

公开(公告)日：1986-06-21

申请号：JP25666984

申请日：1984-12-05

Applicant: Nec Corp

Inventor： KAWANA YOSHIE ,  KIMURA JUN ,  KURIYAMA TOSHIHIDE

IPC: C12Q1/00 ,  G01N27/30 ,  G01N27/414

CPC classification number: C12Q1/001 ,  G01N27/4145

Abstract: PURPOSE:To detect the concentration of several chemical substances in a sample solution simultaneously with a high sensitivity, by providing an FE type ion sensor having an enzyme immobilized membrane on the surface of an insulation substrate while an electrode having an enzyme immobilized membrane is provided on the other surface thereof. CONSTITUTION:This composite type enzyme electrode is made up of two ion sensitive FETs (ISFET) and two kinds of enzyme immobilized membranes provided on the top surface of a sapphire substrate insulator. An FE type ion sensor is made up of a p-type silicon substrate layer 2, an n-type high impurities density source region 3, an n-type high impurities density drain region 4, a p-type high impurities density earth region, a silicon oxide film 6, a silicon nitride film 7 and an ion sensitive film. Two precious metal electrodes 10 and 11 are provided on the undersurface of the substrate 1 and a second enzyme immobilized membrane is formed on the surface of one electrode 11. When measuring urea and glucolse, a urease immobilized membrane and a glucose oxidase immobilized membrane are used for the first and second enzyme immobilized membranes respectively.

Abstract translation: 目的：为了在高灵敏度的同时检测样品溶液中的几种化学物质的浓度，通过在绝缘衬底的表面上提供具有酶固定膜的FE型离子传感器，同时将具有酶固定膜的电极设置在 其另一个表面。 构成：该复合型酶电极由两个离子敏感FET（ISFET）和设置在蓝宝石衬底绝缘子顶表面上的两种酶固定膜组成。 FE型离子传感器由p型硅衬底层2，n型高杂质浓度源区3，n型高杂质浓度漏极区4，p型高杂质密度接地区构成， 氧化硅膜6，氮化硅膜7和离子敏感膜。 在基板1的下表面上设置两个贵金属电极10和11，并且在一个电极11的表面上形成第二酶固定膜。当测量尿素和葡萄糖时，使用固定有脲酶的膜和固定有葡萄糖氧化酶的膜 分别用于第一和第二酶固定膜。

公开(公告)号：JPS6188136A

公开(公告)日：1986-05-06

申请号：JP20916684

申请日：1984-10-05

Applicant: Nec Corp

Inventor： KURIYAMA TOSHIHIDE ,  KIMURA JUN ,  KAWANA YOSHIE

IPC: C12Q1/00 ,  G01N27/00 ,  G01N27/414 ,  G01N27/30

CPC classification number: C12Q1/001 ,  G01N27/4145

Abstract: PURPOSE:To form enzyme immobilizing films which are different from each other on the same chip by removing part of the photoresist coated on a semiconductor wafer then coating a protein soln. thereon and dissolving the photoresist. CONSTITUTION:The photoresist film 6 soluble in acetone is spin-coated on the front of a wafer constituted by forming semiconductor field effect type ion sensors ISFETs by using an island-shaped silicon layer on the front of a sapphire substrate 1 and depositing gold 10 by evaporation on the rear. The photoresist film 6 on the surface to be provided with the enzyme immobilizing film 9 is thereafter removed by photolithography using a photomask. A protein soln. contg. the 1st enzyme and crosslinking agent is then spin-coated to form an urease immobilizing film 7. The water is then immersed in acetone to dissolve the photoresist 6 and at the same time to remove the urease immobilizing film coated thereon. A glucose immobilizing film 9 is also formed by repeating the similar stage.

Abstract translation: 目的：通过除去涂覆在半导体晶片上的部分光刻胶，形成蛋白质溶胶，形成在同一芯片上彼此不同的酶固定膜。 在其上并溶解光致抗蚀剂。 构成：可溶于丙酮的光致抗蚀剂膜6通过在蓝宝石衬底1的前面使用岛状硅层形成半导体场效应型离子传感器ISFET而形成的晶片的前面旋涂，并通过 后方蒸发。 然后通过使用光掩模的光刻法去除设置有酶固定膜9的表面上的光致抗蚀剂膜6。 蛋白质溶胶 对比 然后旋涂第一酶和交联剂以形成脲酶固定膜7.然后将水浸入丙酮中以溶解光致抗蚀剂6，同时除去其上涂覆的脲酶固定膜。 还通过重复类似的阶段形成葡萄糖固定膜9。

公开(公告)号：JPH02168153A

公开(公告)日：1990-06-28

申请号：JP32521688

申请日：1988-12-22

Applicant: NEC CORP

Inventor： KAWANA YOSHIE ,  ITO SHIGEFUMI ,  MIYAMOTO SHIGEYUKI

IPC: G01N27/327

Abstract: PURPOSE:To form an immobilized enzyme film with uniform thickness by developing a wafer and a solution of protein while cooling the both to equal to or lower than 15 deg.C when said wafer onto which said solution of protein is developed is rotated to form a thin film. CONSTITUTION:A photoresist solution is developed on a substrate 4 of sapphire having an ion sensor 5 of semiconductor field effect type thereon, and the substrate 4 is rotated at 3,000rpm for 30 seconds. Then, the substrate is pre-baked, exposed and developed with the use of a photomask, so that a part of a photoresist film 6 where an immobilized enzyme film 7 is to be formed is removed. Thereafter, 0.6ml PIPES buffer solution containing, e.g., 60mg/ml glucose oxidase, 15% bovine serum albumin and 1% glutaraldehyde is developed on the wafer. At this time, the buffer solution is 0 deg.C and the wafer is 4 deg.C. The wafer is immediately rotated at 2,000rpm for 30 seconds so that the solution is applied all over the surface of the wafer. After the enzyme film is completely crosslinked through one-hour reaction at 20 deg.C, the wafer is soaked in acetone, whereby the film 6 and the film 7 applied on the film 6 are removed. Thus, the film 7 having uniform thickness can be formed.

公开(公告)号：JPS6186644A

公开(公告)日：1986-05-02

申请号：JP20862684

申请日：1984-10-04

Applicant: Nec Corp

Inventor： KIMURA JUN ,  KURIYAMA TOSHIHIDE ,  KAWANA YOSHIE

IPC: G01N27/327 ,  C12Q1/00 ,  G01N27/414 ,  G01N27/30

CPC classification number: C12Q1/001

Abstract: PURPOSE:To obtain economical manufacturing method of a biosensor characterized by a small amount of consumption of enzyme, by forming a plurality of ion sensitive parts on a chip, coating a part other than the sensitive parts by a hydrophobic resin beforehand, holding a liquid including the enzyme, and forming an immobilized enzyme film. CONSTITUTION:A part on a substrate 1 other than ion sensitive parts 2 is coated by a hydrophobic resin 3. A solution including enzyme 4 having albumin and enzyme is formed on one gate by a syringe. The solution including enzyme 4 loses water and a film including enzyme 5 is formed in several minutes. Then glutaraldehyde is supplied thereon as a bringing agent 6. It is repelled by a resin layer (fluoride) and held on the previously formed protein film. This is made to remain for several minutes. Then the protein bridge is favorably expanded, and a fixed enzyme film 7 is formed. Then the product is made to remain intact for a specified time period. After washing with water and glycine treatment, the product is stored in a buffer. Thus a small amount of the enzyme can be well fixed on a minute sensor.

Abstract translation: 目的：为了获得生物传感器的经济性制造方法，其特征在于消耗少量的酶，通过在芯片上形成多个离子敏感部件，预先用疏水性树脂涂布敏感部件以外的部分，保持包含 酶，并形成固定化酶膜。 构成：除了离子敏感部分2之外的基板1上的部分被疏水性树脂3包覆。通过注射器在一个浇口上形成包含具有白蛋白和酶的酶4的溶液。 包含酶4的溶液失水，并在几分钟内形成包含酶5的膜。 然后在其上供给戊二醛作为引发剂6.它被树脂层（氟化物）排斥并保持在先前形成的蛋白质膜上。 这是为了保持几分钟。 然后蛋白质桥被有利地扩大，形成固定的酶膜7。 然后使产品在一段特定的时间内保持完好。 用水洗涤和甘氨酸处理后，将产物储存在缓冲液中。 因此，少量的酶可以很好地固定在微小的传感器上。

公开(公告)号：JPS6135786A

公开(公告)日：1986-02-20

申请号：JP15612384

申请日：1984-07-26

Applicant: Nec Corp

Inventor： KIMURA JUN ,  KURIYAMA TOSHIHIDE ,  KAWANA YOSHIE

IPC: G01N27/327 ,  C12M1/40 ,  C12N13/00 ,  G01N27/30

Abstract: PURPOSE: To carry out the partial inactivation of an immobilized enzyme membrane, in high efficiency, and to prepare a partially inactivated immobilized enzyme membrane, easily, by irradiating a specific part of the immobilized enzyme membrane with electromagnetic wave to inactivate the enzyme of that part.
CONSTITUTION: A specific part of an immobilized enzyme membrane is irradiated with electromagnetic wave such as ultraviolet radiation, X-ray, infrared radiation, microwave, etc. to inactivate the enzyme of the part and to obtain a partially inactivated immobilized enzyme membrane. For example, a differential enzyme electrode can be produced with extremely simple process, by forming an immobilized enzyme membrane on an element having plural ion-sensitive parts, shielding the immobilized enzyme membrane partly with a mask made of chromium, etc., and irradiating the immobilized enzyme membrane with a low-pressure mercury lamp, etc. emitting light shielded by the light-shielding part of the mask and having wavelength active to inactivate the enzyme, thereby effecting the partial inactivation of the enzyme.
COPYRIGHT: (C)1986,JPO&Japio

Abstract translation: 目的：为了高效率地进行固定化酶膜的部分失活，容易地通过用电磁波照射固定化酶膜的特定部分来使部分灭活的固定化酶膜失活，从而使该部分的酶失活 。 构成：用紫外线，X射线，红外线，微波等电磁波照射固定化酶膜的特定部位，使部分酶失活，得到部分灭活的固定化酶膜。 例如，通过在具有多个离子敏感部件的元件上形成固定化酶膜，通过用铬等制成的掩模部分地遮蔽固定化酶膜，可以以极简单的方法制造差示酶电极，并照射 发射由掩模的遮光部分遮蔽的光并具有波长活性以使酶失活的低压汞灯等的固定化酶膜，从而实现酶的部分失活。

公开(公告)号：JPS6410163A

公开(公告)日：1989-01-13

申请号：JP16634287

申请日：1987-07-02

Applicant: NEC CORP

Inventor： KAWANA YOSHIE ,  ITO SHIGEFUMI

IPC: G01N27/30 ,  C12Q1/00 ,  G01N27/28

Abstract: PURPOSE:To measure the concn. of a chemical material in a minute amt. of a sample by using a microsensor by providing a sensor part to one end of a circuit board having elasticity and providing a piezo-electric actuator to the other end. CONSTITUTION:The tip of a glucose sensor chip 3 constituted by forming a glucose oxidase immobilized film in the sensitive part of a field effect type ion sensor is inserted into a drop 4 of the sample liquid dropped onto a fluoroplastic substrate. The sensor chip is adhered to the flexible printed circuit board 2 formed of an epoxy resin and the laminated piezo-electric actuator 1 is installed to the upper part of the printed circuit board. The oscillation of this actuator 1 is amplified by the elasticity of the circuit board 2 and is transmitted as a large displacement to the chip 3. While the soln. is efficiently stirred by the vibration, the concn. of the chemical material in the soln. is measured.