JP2527933C - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a device for use in a chemical (particularly biochemical or clinical) test method,
Fabrication method and use of the device. In one embodiment, the device detects and measures inorganic ions in an aqueous sample
And, in other embodiments, specific binding assays.
Intended. Conventionally, from many other liquid containers for assay reactions, conventionally about 0.5 ml
Using a so-called microtiter well with a working capacity of
Have often implemented microscale methods. Micro-scale assembly
Other devices and devices for manipulating materials are described, for example, in EP 031993,
Described in the specifications of National Patent No. 1571772, British Patent No. 1584129 and British Patent No. 1414479
Have been. In particular, the prior art contains numerous disclosures of analytical devices for manipulating and measuring small test samples. British Patent No. 2 090 659 [I
Instrumentation Laboratory, Inc.
Are self-filling metering channels, and
It is constructed with a lip or inlet that can hold a sample of about 10 μl
Pick up (for example) 10 μl by elephant and remove the fibers on the filter layer below the transparent window
A test strip to be reacted with the reagent contained in the ionic pad is described. Result is
For example, it can be seen with the naked eye as a color reaction. UK Patent No. 2 036 075 [HE Mennier], British Patent
Patent No. 1 104 774 [JP Gallagher], European Patent
No. 0 057 110, No. 0 034 049, No. 0 010 456 [Kodak (K
odak)] are all capillary channels or channels for manipulating biological or test fluids.
Describes other aspects of the use of chamber size. The prior art also contains many disclosures about chemically sensitive or selective electrodes.
Included, `` Ion-Selective Electrode in Analytical Chemistry ''
s in Analytical Chemistry). H. Freiser, Plenamp
Les (Plenum Press) 1978). According to the invention described herein, a capillary-filled cell device that can be easily manufactured
Chairs provided, electrically monitored tests using very small liquid samples
Facilitates, for example, specific ion analysis, enzymatic reactions, and specific binding assays
. According to the present invention, Each A cavity is used to store sample liquid by capillary action.
One having a size small enough to flow into the cavity Or multiple The mold
The cavity has one or more electrically measurable properties in the sample.
The electrode structure for measuring the
Consisting of conductive coating and cavity supports specific reaction of sample
Specific countermeasures, including a coating of material necessary to perform
A device is provided for collecting and testing specifically-reactive samples. According to the present invention, (a) (usually) a sheet to provide a number of device parts
Forming a coating on the surface of the material, (b) together with the coated sheet material,
Each device of the device provides an additional capillary-sized cavity for collecting and holding an amount of sample fluid by capillary action in contact with the reactive coating
Forming a structure, (c) forming an electrode structure within each cavity, and (d) each one or
Step of separating the sheet material into sections providing multiple sample collection and test devices
Also provides a method for collecting specific reactive samples and manufacturing test devices
I do. First, on the surface of the carrier which later constitutes the wall of the capillary cell, as in the example below.
Form a conductive layer, then add a layer of any auxiliary material, and finally a cell or some
It has been found that it is usually convenient to aggregate these cells. The coating is a buffer that must be released into the liquid sample material incorporated into the device
Agent, or a specific reagent such as a protein-containing binder or enzyme.
It may be a responsive substance. Coating can be fused or continuous, or
For example, the pattern is divided into separated patterns, such as a two-dimensional array of patches.
It may be. Device to form part of the capillary cell cavity
Including an ion selective membrane coating over a preformed or coated electrode on a wall
Good. When making these patches, first create a continuous coating and then, for example,
Either remove that part to leave the desired pattern, such as an array of parts, or
Can be produced by inactivation. The coating provides specificity for the desired assay
It can be a membrane of a releasable reagent having, for example, a releasable antigen or antibody or its attraction
Conductor coating Or, for example, An antigen or antibody covalently linked to form an immunosorbent or
Can be an immobilized specific binding substance such as a derivative thereof. Completion of capillary cell
Additional structures that allow, for example, preferably a defined reproducible volume of sample liquid
Capillaries less than about 1 mm, for example, that can be trapped between sheets by tube action
Separated from the first sheet in a capillary space, with a suitable bonding adhesive
Thus, it may be another sheet material bonded to the first sheet material. example
Sheet material, such as glass, quartz or plastic material, is drawn with a line
Can be divided into units by tearing or sawing or cutting.
In the following examples, the sample liquid can be loaded or added thereto, and
An external loading surface or opening that flows into the vise cavity
It is implemented to leave the department. The external loading surface at least defines the cavity
Contains sufficient liquid (eg, in the form of droplets of a substance spreading on a surface) to sufficiently fill or
Preferably, it has the ability to hold. In the embodiment of the device as shown in FIG. 1 below, for example, the second sheet
The sheet with the electrodes extends beyond the end of the capillary cell
It is extended and provides an anchoring point for electrical connections to external circuits.
You. According to the present invention, specific reactive sample collections produced by the methods described herein are provided.
A collection test device is also provided. The fabrication of the capillary cell electrode structure of the present invention may be performed in any of a number of ways. For example, on a glass, silica or plastic substrate to form the walls of a capillary cell
The conductive coating of tin oxide SnO2, in particular the electrode pattern, is left in a manner known per se.
It is also possible to provide a material which has been partially etched and removed. like this
On the conductive electrode, any reactive or ligand described elsewhere herein.
Overlying the coating associated with the metallization. Alternatively, the glass, silica, crystal, ceramic or ceramic to form the walls of the capillary cell
Each form an electrochemical half-cell on the surface of the plastic
One or more conductive coatings, such as a coating forming a silver-silver chloride electrode.
These coatings may optionally have a top layer of ion selective or other chemically selective membrane.
May be. Examples of electrode structures to be included in a capillary cell according to embodiments of the present invention are
The matrix incorporates conductive particles into a non-conductive matrix (eg, polyvinyl chloride).
A composite electrode composed of a polymer body such as Nyl). Conductive particles include particles
It is such that it can provide a conductive connection from the composite to an external electrical circuit. Below
In the example shown, the conductive particles have a conductivity corresponding to a resistance of a few milliohm-cm.
To provide a composite electrode. This value is by no means critical and can be several orders of magnitude
Can be used, or the resistance of the electrode material is generally the resistance of the external circuit (often
(Several megaohms). Such an electrode structure is also made of a non-conductive polymer material (capillary cell).
(Forming the walls of the shell). Non-conductive solid matrices or other materials that can form other substrates other than polyvinyl chloride
The material is, for example, polyurethane, polystyrene, polyvinyl acetate, epoxy resin
(Especially as a matrix of conductive particles) and methacrylate plastic
And an inorganic matrix such as glass. Hereinafter, polyvinyl chloride (PVC)
The description states that any of these alternative matrix materials can be used.
I taste. In the structure provided by the present invention, the conductive particles may be, for example, graphite, silver,
, Platinum, gold or copper. If desired, the machine incorporating the particles
The trick is to conduct conductive tracks on a flat insulating substrate, such as a ceramic substrate.
Thick-film microcircuit manufacturing for painting or screen printing
Used as a base for conductive paints, known and useful for use in
It can be an organic matrix obtained by solidification from a paint vehicle.
The preferred particle size of the conductive particles can be in a wide range, for example (on the order of 10-20 microns)
From 200 microns (on the order of). Of particle incorporation into the complex
Suitable levels are, for example, of the order of 10 to 80% by volume (based on the composition volume), for example 50%.
Including the volume% level, or depending on the nature of the particles, to reach the desired electrical conductivity.
May include required levels. In one example of an electrode structure made of silver-PVC, (maximum particle size)
50 micron silver particles, 3: 1 silver particles: PVC particles by weight. One preferred form of electrode is a conductor and in direct contact with the conductor
Non-conductive membrane matrix material (eg, glass or organic polymer)
Ductors consist of a matrix that incorporates conductive particles as described above.
The material is an ionophore to sensitize the electrode to the particular analyte to be studied.
The organic film material consists of a conductive complex.
It is tightly bound to the matrix. For example, membrane material and matrix are convenient
May be the same polymer, for example in a simple method such as solvent welding.
Either can be melted or bonded together. When using PVC as a matrix material for conductive particles,
Use PVC that contains no or very little plasticizer.
(For PVC in the region containing the conductive particles). (We call this "
Marked "pure" PVC. ) (Plasticizers in the range of 10% by weight or less exist in this region.)
It is preferably present, preferably less, for example <5%, for example <1%. For membrane materials, examples of suitable contents of ionophores or other sensitizing components are, for example, membranes
It is in the range of 5-10% by weight of the total composition. The thickness of the ionophore-containing region is comparable
0.1 to 0.5 mm. PVC in areas containing ionophores or other sensitizing components
Is usually more conveniently used as in conventional ionophore PVC membranes.
Should contain relatively large amounts of plasticizer, such as many ionophore-containing layers
May comprise, for example, a PVC: plasticizer composition in a weight ratio ranging from 1: 1 to 1: 2.
. Examples of useful plasticizers for use with ion selective polymer membranes, especially PVC membranes
Is dioctylphenylphosphonate, diethyl adipate, dioctylsebacate
G, trioctyl phosphate and o-nitrophenyl phenyl ether
It is. An example of a useful ionophore is calcium diisooctylphenylphosphine.
(For calcium sensitive electrode), valinomycin (for potassium sensitive electrode)
, Tridodecylamine (for hydrogen ion sensitive electrodes), chloride, bromide or silver iodide grains
(For halide-sensitive electrodes) Silver sulfide particles (for sulfide-sensitive electrodes), sulfide
Contains a mixture of silver and copper sulfide particles (for electrodes sensitive to sulfide and copper)
In general, any conventionally used in crystalline form to make single crystal electrodes
The finely divided granular form of the material is converted to a polymer or other nonconductive matrix for the electrode membrane.
Liquor, the amount of which corresponds to the surrounding solution to be tested or measured
Particles make an electrically effective contact so that the electrode can react to the presence of components that
Is enough. The capillary cell device of the present invention can also have the following features in one embodiment. (a) if desired, at least one of the walls surrounding the cavity is visible and / or ultraviolet
It can be transparent to light, such as lines, and has an optically uniform and generally smooth surface.
This allows for in-situ sample collection as well as electrical measurements possible with electrodes.
Measurement and / or optical analysis of a reaction product having specific binding ability
be able to. (b) In some cases, the cavity of the device has two opposite sides forming a cell.
Between thin walls, preferably made as a combined or integrated unit
It can be bitty. In some cases, for example, such an embodiment may be used as an unfilled liquid crystal display device as obtained as an intermediate stage in the manufacture of liquid crystal displays.
It is possible to include a connection structure of a plate similar to the structure of the vise. According to one aspect of the invention, (preferably disposable) (possibly translucent or transparent)
Are provided, which can be manufactured by the methods described herein,
For performing a combined assay, with a spacing of less than about 1 mm and a certain (preferred)
Or specified) amount of (usually aqueous) liquid that is taken up and retained by capillary action.
To form a liquid-retentive cell with an opening
Consisting of a set of facing plates that are
Include suitable enzymes, dye molecules, antibodies, antigens or buffer salts for the test to be performed.
Having a membrane and for measuring one or more electrically measurable properties of a sample
Includes electrodes or electrode structures. "Defined medium" is the shape of the cell itself
This is a volume that is substantially determined by the shape and arrangement.
It is not clearly determined from the quantity. Cells of the above type can be assembled from sheets of glass or plastic.
When using plastic sheets, be sure to use the correct mold, for example a capillary cell cap.
With spacers like ridges to adjust the space of the wall of the component of Vity
Can be in the form of something. A sufficient amount of sample can be applied to fill the cell, from which capillary action
An outer surface portion where the sample can easily flow into the capillary cavity or
The cell can have a lip. Large enough for convenient loading of samples
With the cell aperture facing outward a sufficient distance to provide a
By extending one of the plates, such a helicopter can be easily formed.
Wear. Another shape of the inlet is formed by an opening in one wall of the capillary cell
For example, a hole for exposing a portion of a wall facing a cell for loading a sample.
It is. Porous filters, such as porous plugs in helicopters or openings
Or, for example, by attaching a filter paper or dialysis membrane to remove that portion of the sample entering the capillary cell.
Filtration or dialysis can be performed to the desired degree. The cell is sealed using epoxy resin, leaving an opening, for example, a rectangular capillary cell.
This can be done by lining the resin along two opposite sides of the capillary, and another one to allow air to escape from the capillary cell as the loading opening and cell are filled.
Make one opening. Preferably, the solid particles sink onto the resin, so that the desired
The resin may include solid particles to secure the space. Capillary tube selected
A substantially monodisperse Balotini (mono) corresponding to the gap or about 100 microns in diameter, etc.
disperse ballotini) (fine glass particles) or, for example, 8 micron in diameter
Ron, 50-100 micron long short glass fiber (e.g. long glass fiber
Motor grind the filter and remove the remaining long fiber by sieving.
A small space on the order of the diameter of the barotini or the fiber.
Suitable for conditioning. Generally, as a non-limiting example, a sky in the range of 5-500 microns
The interval is selected. Fibers are less than about 50 microns in diameter than monodisperse barotini
Select a fiber for a very narrow gap because it is easy to obtain. Wider gap
Is preferably barotini. The substance or one of the substances forming the coating on the walls of the capillary cell cavity is, for example,
It can be an enzyme, antigen or antibody. Preferred examples are urease, glucose oxidase
Or concanavalin A or anti-globulin antibody. Especially in the case of enzyme coating
The enzyme can be immobilized or coated in a releasable form. like this
Such a protein may be glass or silica or any other method performed for immobilization.
Can be immobilized on plastic surfaces. For example, simultaneously or sequentially
It is useful to simply coat the binder and sucrose on the rear surface and dry. If desired
For example, especially in the case of plastic materials, European Patent No. 0 014 530 [Unili
(Unilever)] and any of the methods described in the references cited herein.
For a wide range of carrier materials, including glass and quartz-containing materials such as silica.
`` Immobilized Enzymes for Industrial Reactors
actor)] [Messing, Academic Press, 1975
: In particular Filbert Chapter 3) or, for example, in US Pat.
Conducts sharing and other immobilization in any of the ways described in GB 1530997.
I can. Particularly in the case of the following examples of electrodes for measuring a potential difference or for measuring a current, a capillary cell
The substance that forms one or more films on the surface of the can be an ionic salt, such as a buffer salt, for example, a glaze-forming inert such as sucrose or other non-ionic wetting agent.
It may be a thin coating of salt premixed with the substrate. Especially in the case of an electrode for current measurement, the electrode structure is
Mediators that facilitate the transfer of electrons to the conducting moiety, such as glucose oxida
Coated with ferrocene, which can facilitate the transfer of electrons from the
It may be. The area between the two electrodes on one wall of the cell is used in certain immunoassays.
Coated with a specific binder that can bind to conductive particles such as gold sol particles used
be able to. The embodiments of the present invention are described by way of example with reference to FIGS.
I do. FIG. 1 shows the present invention. According to the embodiment of Of capillary cell device containing electrodes
FIG. Figure 2 Another Schematic of disposable capillary cell device
The cross-sectional view is shown schematically. FIG. 3 shows a schematic diagram of the cell device of FIG. 2,
2 includes a line II for indicating the line of the cross section of FIG. FIG. 4 shows a schematic fragmentary view of an intermediate stage of manufacturing a plurality of the devices of FIGS.
You. 5 to 8 show further examples of the present invention. other Schematic side view of the electrode device provided in the embodiment
FIG. FIG. 1 shows the shape of the capillary cell device of the present invention. Cell shown in Fig. 1
The device comprises an upper plate separated from a lower plate 2 by a coupling track 3
Consists of one. The three multiple capillary cavities are spaced apart from each other by
It is formed by four connecting tracks 3 delimiting the sides of the tee. Fig. 1
The vise has an interdigitated electrode structure consisting of electrodes 10 and 11,
Each one has an overall oxide mask that has been etched away to form the pattern shown.
(SnO2). The conductive tin oxide layer is semiconductive
In a manner known per se in the body and liquid crystal industry, and not itself a part of the present invention,
Manufactured and etched. If desired, one or more coating layers (not shown in FIG. 1) can be formed on top of the tin oxide coated electrode structure and / or on the inner surface of the facing plate 2. The electrodes 10 and 11 continue beyond the capillary cell to the surface 6 of the plate 2 and to another electrical circuit.
Ends at connection points 12 and 13 for connecting to roads and devices. (If desired,
The poles 10 and 11 are passed under the bonding layer 3 to appropriate connection points other than those shown in the drawing,
It may continue up to the connection point of another part of the cell. ) Each component cell of the device has one set of electrodes 11 and 12. Only one set is complete
They are fully illustrated and are fully referenced in the figures. With this device, the platform
Form 6 is not used for sample loading and plate 1 on the other side of the device.
Plate 2 also extends beyond to provide a sample loading platform
I have. Combined track to divide the platform into three sample loading areas 15
This platform is separated by an extension 14 of the two central tracks of
You. The three sets of connectors 12 and 13 are the edges of the plate 2 having the connection points 12 and 13.
Edge-connector device that can fit the entire capillary cell device
It is adjusted to be compatible with the corresponding wire 16 of the chair 17. Such capillary cell devices are useful for measurement, and in some cases
Other conductive materials, such as conductive particles, such as gold sol particles coated with a mechanical binder
Can be included. 2 and 3 illustrate the implementation of the present invention, except that the electrode structures are omitted from the drawings.
Aspect of the device Capillary cell Is shown in a schematic form. Device is easy to operate
The size is, for example, 3 cm × 1.5 cm. The device is located on top (eg plastic, gas
(Las, PVC or silica) plate 1 and lower (for example, the same) plate 2 (about 1 m)
m thickness), which are applied to the bonding track 3 of a suitable (eg epoxy) adhesive.
It is fixed together so that it faces in parallel at a distance of less than 1 mm, and both ends are open.
Forming a capillary cell cavity 4 which is
The outside of the plate 3 is connected to the outside through the first
To form an opening. Another discontinuity at the other end of the connecting track 3
And another opening to allow air to escape when the sample liquid fills the cell.
Mouth. The plate 2 is larger than the plate 1 and has a portion 6 extending from the opening.
doing. Portion 6 of plate 2 acts as a platform or threshold or helicopter to which sample droplets can be applied, so that the capillary flow
This liquid can fill the capillary cell cavity 4. Cavity 4
Attracts and contains a defined and appropriately reproducible amount of liquid when filled as in The layer 7 of the substance relevant to the test method in which the capillary cell is to be used is applied to the inner surface of the capillary cell.
Immobilized on Do . In the embodiment shown in the drawing, layer 7 is a patch of material on plate 2
Yes, on other devices, on plate 1. Tests based on enzyme activity, such as urea
The purpose of the measurement of urea measurement by conversion to ammonia using
It can be a region of an immobilized enzyme such as an enzyme. Also in connection with immunoassays
The layer can be, for example, an immobilized antibody. There may be one or more such layers, for example,
For example, there may be one layer on plate 1 as well as plate 2, or
Multiple layers may be stacked and / or side-by-side on the sheet. Fig. 2-3
Although not shown in FIG. 1, the layer 7 or other layer lining the inner surface of the capillary cell is shown in FIG.
And one or more conductive layers as described with respect to FIG.
(Not shown), a conductive external connection is made between the bonding layer 3 and the surface of the plate if desired.
Through conductive tracks or connectors from inside the cell to outside the cell
Provided. These are often used in the manufacture of semiconductors and liquid crystal displays
Manufactured in a manner known per se used for conventional surface treatment of such conductive tracks
Is done. The section shown in FIG. 2 is empty because the line does not extend through the connecting track 3.
Shown are plates 1 and 2 with a space between them. The production of a plurality of cells as shown in FIGS. 2-3 is shown in FIG. Figure 4 is like this
FIG. 4 is a fragmentary view showing an intermediate stage of manufacturing a simple cell. Glass or other to make plate 2
A large plate 8 of material of any kind is cleaned and, in a suitable way,
The electrode layer is covered with a patch of material 7 and the
Be converted. A second plate (not shown) may have a connecting track corresponding to track 3 as appropriate.
After forming the rack in the soil and with any other desired material,
After appropriately forming a hook, it is pressed against the plate 8 to solidify the adhesive. Next, the fourth
The line indicated by the dotted line 9 in the figure and the corresponding line on the upper plate (however, labeled as line 9)
Break or cut along the line). As a result, a cell like the cell shown in FIGS. 2 and 3 is obtained. The apparatus of FIG. 4 is described with respect to the use of glass sheets as a substrate method.
Use, especially but not exclusively, plastic sheets.
It is convenient to use a spacer ridge other than
Before assembling the capillary cells, open the inlet as described elsewhere herein.
Adds inlet apertures and filter devices as part of such sheets
You can also. Capillary cell device of FIGS. 2-3 among other devices made according to the invention
Is equipped with a convenient-shaped handling-piece or holder, if desired.
For this purpose, if not formed in one piece with the cell device,
Fixed or removable for use with such holders
Any convenient form of connection device may be provided. The device includes a thin coating layer of releasable biochemical reagent, for example, a pre-
Air-drying the protein-sucrose mixture in a thin film on a plate.
No. These are selected according to the specific test chemistry to be performed in the device.
, Are assembled. The range of chemical or binding reactions that are part of the test to be performed
Are all types of electrochemical, enzymatic, binding and quenching reactions
However, some studies have shown that such reactions occur in a capillary cell.
Emphasize that they are not necessary. In addition to the releasable coating layer, the device grows
It may include an immobilized coating layer of a biological reagent. Formation of a reactive immobilized protein layer (eg, enzyme or antibody) on the inner surface of the cell
For example, it can be performed as follows. For example, about 1 mm thick, large enough to include a two-dimensional array of cell parts, multiple
A sheet of glass (eg soda) with several cell units of
Suitable methods such as washing and sonication and, if necessary, solvent evaporation in known manner
By oil removal by air or continuous with ammonia hydrogen peroxide and hydrochloric acid / hydrogen peroxide
Temperature treatment (80 ° C), washing with water and air drying at, for example, 115 ° C for 30 minutes.
You. Next, a patch of the desired protein or other coating is obtained in the following or an equivalent manner.
Apply the pattern. First, a known method (for example, terminal amino-alkyl tri-
Methoxysilane, or other reagents substantially, such as 3-aminopropyl compounds, substantially U.S. Pat.
Using another reagent described in US Pat.
Reaction of glass with silane-based coupling compounds (at about 2% v / v in tonnes)
The amino terminal immobilized on glass is then glutarized (eg, 2%, pH 7).
React with aldehyde, remove excess reagent, and solidify by known component methods.
Exposure of glass activated with immobilized aldehyde groups to reaction with proteins in solution
This achieves covalent attachment of the antigen or antibody or other protein. For example,
Here, it has been found that treatment at 37 ° C. and about pH 9.5 for 2 hours is suitable. Gala
A suitable final active protein loading rate on the surface is, for example, about 0.5
μg / cm ^Two It is. This constitutes a continuous or near-continuous layer
it is conceivable that. The amount or density or specific activity of the immobilized layer may vary depending on the specific assay chemistry.
Determined by susceptibility requirements, it does not itself form part of the present invention. For example, strong
Wash with strong buffer (0.1 M acetate, 0.5 M NaCl, pH 4-5), then wash with neutral buffer
(PH 7-7.4), then wash with pH 9-10 and remove excess reagent by neutralization
it can. Later etching or inactivating any part of the protein (eg enzyme coating)
If one desires, the following approach can be used. The coated sheet is then substantially aerated.
Can be placed in a closed atmosphere draft that does not contain, for example, another inert
Air gap on the coated side placed close to the surface can be reduced to less than about 1mm
. Next, the pattern corresponding to the portion to be removed or inactivated by etching the coating (for example,
The sheet using an image patterned with ultraviolet light
(It is preferable to use light in a narrow wavelength band that can be implemented around C.280 nm.)
), Leaving a pattern of the remaining active protein patches. For example, a few
Irradiation can be performed using a separated GE7-Watt mercury lamp for about 5 to 20 minutes.
it can. With masking or real imaging system close to the plate
Irradiation patterns can be created. The UV etching used here is
JA Panitz and I Giaver are shining
Surface Science, 97 (1980) pages 25-42
It is thought to be based on the same principle as the UV etching process. Next, the desired pattern is formed to form a connection with the overlying plate.
UV curable epoxy adhesive printed on a glass plate patch-coated with epoxy
I do. Silk screen techniques that are customary in themselves and are not part of the present invention
Apply xy glue. Add a small amount of epoxy resin to the resin (for example,
Made by removing long fibers left after sieving and sieving), about 20 micron in diameter
Ron may have short glass fibers of about 100-200 microns in length. The
Use barotini in epoxy resin instead of lath fiber pieces as follows:
Is also preferable. For example, to create a gap of 100 microns, the corresponding size
Incorporate the barotini in the epoxy. Slightly thicker than desired space between plates
For example, if you want a 10% thicker space, say 100 microns,
Apply the epoxy layer by screen printing and gently press the additional plate into place
Epoxy can be spread lightly. If desired, patched with the same or a different protein or other coating material
A coated or uncoated second similar glass sheet is patterned as
A mirror image of the first pattern of epoxy adhesive can be applied, and then the two
Vacuum or deoxygenate if necessary to solidify turn sheets
, And is solidified by ultraviolet irradiation. Coating protein or other material that should remain active
Apply UV light on the image with the pattern to avoid patches. After solidifying the adhesive, any known method used in the manufacture of liquid crystal devices,
CH-627559 and 629002, especially related to the production of liquid crystal display devices
Break two plates by drawing lines to individual cell units in the manner referred to in the book
(scribed and broken). The correspondence between these specifications and the method of the present invention
The steps can be performed in a similar manner, mutatis mutandis. A convenient form of cell obtained by this process is that the capillary cell takes a defined amount of aqueous solution.
In order to be able to lift up, the incomplete frame of the bonding material
Having an air-separated space of at least about 5-500 microns (at least
(It can also have one opening and another opening for air to exit)
And two substantially parallel oriented layers. One of the glass layers extends beyond the opening of the cell and places a droplet on its surface, causing it to be totally or partially within the cell.
Can be put in. In particular, devices made of plastic materials
An opening can be made or left in one wall of the cell to load the sample.
It preferably has the above-mentioned filter device. 5 to 9 show further examples according to an embodiment of the present invention. 5 electrodes Containing capillary selde
The vise is shown schematically in cross section. In each drawing, groups 51 and 52 are labeled
Other structures besides the electrodes for clarity.
Has been torn. In each case, it is convenient if the capillary gap is on the order of 0.1-1mm
May be useful. In FIG. 5, a pair of electrodes 55 and 56 which are spaced apart from each other are fixed to the surface of the wall 52.
It is shown as a layer. The electrode 55 has an ion selectivity (for example, potassium ion selectivity).
A) electrode 56, which effectively acts as a reference electrode, eg a thin layer of polystyrene
An ion-insensitive electrode such as an electrode coated with. Appropriate mode of electrode configuration
Are described below. Fig. 6 shows a potassium ion consisting of two electrodes 61 and 62 separated by a space.
1 shows a capillary cell device containing electrodes for measuring the concentration. 61 (Kariu
62) is a pH-sensitive reference electrode. Facing capillary cells
A releasable layer 63 containing a pH buffer (containing no potassium) is applied over the wall 51.
Overturn. This coating is released into the sample liquid when taken into the capillary cell.
A releasable coating that is dissolved (eg, sucrose grise). In FIG. 7, the electrode 62 is sensitive to ions (eg, chlorine) (reference ions) different from 61.
The electrode is a highly selective ion-selective electrode, and the buffer film 63 absorbs the salt of the reference ion.
Contains an impaction amount, for example, more preferably non-
A sump introduced into the capillary cell to always provide a high chloride ion concentration
That contain chloride ions that should be released and dissolved at standard concentrations in
The electrode 61 is a device similar to the device of FIG.
If not, it is convenient if the cationic component of the salt in the coating 63 is calcium.
possible. FIG. 8 shows another device for measuring potassium ion concentration. In FIG. 8, electrodes 81 and 82 are the same ion selective electrode, in this case potassium selection.
It is a selective electrode. The releasable coating 83 on the opposite wall of the cell is incorporated into the cell.
Standard concentration of potassium to be released into the sample solution to be released (eg as KCl)
It contains. The coating 83 faces the electrode 82 and does not face the electrode 81.
Target or patch-like coating, with the electrode 81 being released from the coating 83 to the electrode 81 during the test
Selected to minimize significant diffusion of ions, eg, significantly larger than 1 mm
A small distance from the electrode 82 and the coating. Another embodiment of the device of the present invention Has an arrangement similar to that shown in FIG.
And an ammonia ion selective electrode or pH electrode and immobilized urease enzyme.
The purpose is to measure the urea concentration using In this example, one of the above mentioned and cited
Select an appropriate substance from the general method and change the electrodes 81 and 82 to an ammonia ion selective electrode.
The coating 83 consists of a layer of immobilized urease enzyme. This capillary tube
When using the urea measurement method with
It is convenient to react the urea content with urease in the sample solution area adjacent to 82.
It can be advantageous. The distance between 81 and 82 depends on this gap during the required reaction time.
Must be large enough to avoid significant ammonia diffusion along
No. Suitable ion-sensitive electrode structures, barriers and coatings may be
The inner wall of the capillary cell structure may be contained on the surface to be formed. For this application
A preferred substrate material is PVC. Screen that can adjust the pattern to apply the material
It is preferred to apply the electrode and coating material by a method such as printing. For example, in order to produce such a structure, various additives are used, for example, as follows:
To create a continuous layer of matrix, such as polyvinyl chloride, loaded with
This is convenient and falls within the scope of the present invention. (E.g. powder suspended in a suitable solvent
Suegin, (eg Johnson Mattey P230 screen pudding
Using a solution of a plasticizer and PVC, including those commercially available as recyclable silver)
A layer of silver-particle-added PVC can be applied as the conductive coating layer. This
It is accessible at its surface and in conductive contact with useful conductive structures
To obtain a high silver chloride content. Also, and under certain circumstances,
Preferably, another zone made of PVC to which silver chloride particles are added is provided with an equivalent effect.
be able to. After treatment to give silver chloride, an ionophore or other sensitizing component
To provide another area of plasticized PVC. Complex
Air-dry, valinomycin ionophore and dioctylphenine in a suitable solvent
A method of applying an ion-selective PVC membrane using a mixture of lephosphonate plasticizers
For example, a potassium ion selective membrane can be formed. Melt (heatable) with appropriate additives
(Plastic) by solidifying the applied layer of PVC, or such as tetrahydrofuran
Coupling by evaporation of the solvent from a suitably added PVC solution in a compatible solvent.
Continuous areas can be deposited, including dip coating, screen printing, spin coating and spin coating.
It can be applied in any suitable manner selected from techniques known per se, such as play.
Can be. Can be manufactured to form part of a composite structure that constitutes an embodiment of the present invention;
The nature and type of ion-selective membranes known per se are very extensive,
It depends on the particular end use to be used. (In connection with conventional wire cores)
GJ Moody and JD Thomas
R Thomas), "Ion Selective Electrodes i
n Analytical Chemistry), edited by H Freiser, plenum pl
(Plenum Press) 1980, Chapter 4 “Poly (vinyl chloride) matrix membrane ion selection
Selective electrode [Poly (Vinyl Chliride) Matrix Membraue Ion-Selective Electrodes]
And U Fiedler and J Ruzika
cka), anal. Him. Acta (Anal.Chim.Acta), 67 , 179 (1973)
All of the membrane electrode compositions are specifically included herein for reference. Details of the manufacture of another membrane included herein for reference are provided by
H. Tamura et al .; Anal. Chem. (1982); 54 , 1224
~ 1227 pages. For example, by screen printing electrodes on one or both of the inner surfaces,
An apparatus for measuring current in a capillary cell can also be manufactured. For example, printing one gold or platinum electrode and one silver / silver chloride electrode
A cell that is more sensitive to hydrogen peroxide is obtained. When a certain voltage is applied inside the electrode
A current proportional to the concentration of hydrogen peroxide in the sample is obtained. Hydrogen peroxide is consumed
As the current decreases, the concentration of hydrogen peroxide in the sample
The first current can be used. By applying the opposite voltage, the oxygen concentration in the sample can be measured. Diffusion limiting layer is deposited on the electrode
Can be useful. Using the cells described herein, a wide range of analytes and reactions, such as ions, enzymes,
Element and enzyme substrate (especially glucose, urea or creatinine)
Can be Many modifications and changes can be made to the invention described in this specification within the scope thereof, and
Any one or more of one or more of the description and accompanying drawings and their equivalents may be made.
Ranging from using.

Claims (1)

Claims (1) A device for collecting and testing a specific reactive sample having a cavity having a size small enough to allow a sample liquid to flow into the cavity by capillary action, wherein the cavity is a sample. An electrode structure for measuring one or more electrically measurable properties of the sample, the electrode structure having a conductive coating on the inner surface of the cavity, and wherein the cavity is unique to the sample. Said device comprising a coating containing a releasable material required to support a reactive reaction and into the cavity. (2) The device of claim 1, wherein the conductive coating comprises at least one of graphite, silver, platinum, gold and copper. 3. The device of claim 2, wherein the conductive coating forms a pattern of conductive electrodes. (4) The cavity of claims 1, 2, or 3, wherein said cavity also comprises an immobilized reagent retained on the surface of the cavity wall, said reagent comprising a biochemical specific binding agent, such as an antigen or an antibody. device. (5) The device according to any of claims 1 to 4, wherein the releasable material comprises a releasable reagent coating. (6) The device according to any one of claims 1 to 5, wherein the wall of the cavity is made of a glass or plastic material, for example, soda glass or acrylic plastic. (7) The coupling structure of the plates, wherein the plates are joined in a parallel relationship with a space therebetween so as to leave a thin flat cavity of a capillary size between the plates.
Any of the devices. (8) The device according to any one of claims 1 to 7, wherein the sample liquid can be applied thereto and has an outer loading surface from which the sample liquid can flow into the cell. (9) The device of claim 1, having a printed pattern of electrodes on the surface of the capillary cell wall. (10) The device of claim 1, wherein said surface is a surface of a solid transparent material so that the contents of the capillary cell can be measured optically. (11) (a) coating a piece of substantially rigid sheet material to form part of the plurality of devices with a conductive coating to form an electrode pattern for the device; (b) substantially rigid The same piece of sheet material forms a coating on another piece containing the necessary and releasable material to support the specific reaction of the collected and tested sample, forming a pattern of discrete portions on the sheet material. (C) forming a capillary tube from the sheet material containing the strips such that the electrode pattern provides a respective electrode structure within each cavity and the pattern of discrete portions of the coating provides respective portions within each cavity. Forming structures that provide cavities for each device to collect and hold an amount of liquid by action; and (d) portions each including at least one sample collection and test device. The structure comprising the step of separating the method of collecting and testing devices of a specific reactive sample for measuring one or more electrically measurable characteristics of the collection and each sample of the liquid sample. 12. The method of claim 11, wherein said conductive coating comprises at least one of graphite, silver, platinum, gold and copper.