WO2000058721A1 - Puce d'electrophorese et appareil permettant de faire fonctionner ladite puce - Google Patents

Puce d'electrophorese et appareil permettant de faire fonctionner ladite puce Download PDF

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Publication number
WO2000058721A1
WO2000058721A1 PCT/EP2000/002510 EP0002510W WO0058721A1 WO 2000058721 A1 WO2000058721 A1 WO 2000058721A1 EP 0002510 W EP0002510 W EP 0002510W WO 0058721 A1 WO0058721 A1 WO 0058721A1
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WO
WIPO (PCT)
Prior art keywords
channels
channel
electrophoresis chip
separation
cathode
Prior art date
Application number
PCT/EP2000/002510
Other languages
German (de)
English (en)
Inventor
Hans-Joachim Hartmann
Renate Konrad
Jörg SCHIEWE
Rainer Pommersheim
Original Assignee
INSTITUT FüR MIKROTECHNIK MAINZ GMBH
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Priority to AU39633/00A priority Critical patent/AU3963300A/en
Application filed by INSTITUT FüR MIKROTECHNIK MAINZ GMBH filed Critical INSTITUT FüR MIKROTECHNIK MAINZ GMBH
Priority to DE10080771T priority patent/DE10080771D2/de
Publication of WO2000058721A1 publication Critical patent/WO2000058721A1/fr

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502753—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44704—Details; Accessories
    • G01N27/44743—Introducing samples
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44756—Apparatus specially adapted therefor
    • G01N27/44791—Microapparatus
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0829—Multi-well plates; Microtitration plates
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B01L2400/0421—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic electrophoretic flow
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02—Burettes; Pipettes
    • B01L3/0241—Drop counters; Drop formers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502707—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the manufacture of the container or its components
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027—General features of the devices
    • G01N2035/1034—Transferring microquantities of liquid
    • G01N2035/1037—Using surface tension, e.g. pins or wires

Definitions

  • Electrophoresis chip and device for operating an electrophoresis chip are Electrophoresis chip and device for operating an electrophoresis chip
  • the invention relates to an electrophoresis chip made of a plate-shaped substrate provided with a cover plate with separation channels which are made in the substrate as grooves, cathodes being provided at the beginning of the separation channels and at least one anode at the end of the separation channels, and being arranged in rows and columns in a matrix-like manner Sample application areas, in the place of which the cover plate has openings. Furthermore, a device for operating at least one electrophoresis chip is described.
  • micro total analysis systems are increasingly being used for analytical and diagnostic purposes.
  • Such systems enable simultaneous and rapid pretreatment, implementation and / or analysis of a large number of samples.
  • electrophoresis chips For the electrophoretic separation and analysis of DNA, RNA or protein fragments or other electrically charged molecules, plate-shaped components with capillary-like grooves are used as separation channels, which are referred to as electrophoresis chips, as miniaturized systems.
  • Methods for microstructuring for example of silicon, metals, glass or polymers, are used for the production of such components.
  • Inexpensive analysis systems obtained from polymeric materials, in particular by means of molding processes have the decisive advantage that, as disposable systems, they do not have to be laboriously cleaned and thus there are no contamination problems.
  • the chip consists of a glass plate with 48 separation channels as grooves. Each separation channel is connected to a sample feed unit integrated on the chip, which enables the serial feed of 2 different samples, so that 96 samples can be separated with 48 separation channels.
  • the microstructured The glass plate is covered with a polymer film, with 96 holes in an 8x12 matrix arrangement, among other things, making it possible to sample with an 8-fold pipetting machine.
  • the sample supply unit in electrophoresis chips usually has a cross structure, i. H. the separation channel crosses at right angles an injection channel arranged between a waste reservoir and a sample application area. The samples are brought into the area of the separation channel by migration in an electrical field applied between the sample application area and the waste reservoir.
  • the separation channels are arranged between the cathode and anode in such a way that they have the same length.
  • an elastomer film with holes in these areas with a thickness of 1 mm is placed.
  • An electrode array held by a circuit board with conductor tracks is located above this arrangement of micro-structured glass plate and reservoir-forming elastomer film.
  • platinum wires are attached to the board, which protrude into the holes in the reservoirs.
  • the detection of the separated samples takes place optically in an area in front of the anode, where the separation channels run parallel to one another.
  • the possibility is mentioned of providing 96 individual separation channels.
  • an application of 4 samples per separation channel is mentioned.
  • a major disadvantage of these known electrophoresis chips is the number of reservoirs to be filled and contacted, which goes beyond the actual number of sample application areas. This involves complex electrical contacting and control. An integration of the electrodes in the electrophoresis chip is not readily achievable with this configuration and provided that electrical conductor tracks and fluid channels do not cross.
  • the object of the present invention is therefore to provide an electrophoresis chip of the type mentioned at the beginning, the sample application areas of which can be arranged in the grid dimension of a microtiter plate with a large number of wells, for example 384, 1536 or larger, and in which the electrodes are integrated is possible in the electrophoresis chip.
  • a device for operating at least one electrophoresis chip which allows easy handling of the electrophoresis chips as disposable parts, is to be demonstrated.
  • This electrophoresis chip An essential feature of this electrophoresis chip is the exactly one sample application area integrated into the beginning of each separation channel.
  • the sample application area is formed by a widened area of the separation channel and connected to a cathode via an intermediate channel.
  • This electrophoresis chip therefore does not have the sample feed units with cross structure used in known electrophoresis chips.
  • sample amounts in the range from 1 ⁇ l to 100 ⁇ l are applied, which is done by pipetting.
  • a sufficiently small amount of it is brought into the area of the separation channel.
  • a sample of a sufficiently small amount can also be introduced directly into a separation channel widened to form a sample application area.
  • the sample feed units previously used, each with at least two reservoirs, can thereby be saved. This also eliminates the need for electrodes and sample reservoirs. This reduces the number of openings to N sample application areas, which also do not need to be electrically contacted.
  • this enables the electrodes required for the actual separation to be integrated on the electrophoresis chip.
  • the space saving achieved in this way creates the possibility of achieving an even higher density of the sample application areas, and thus also of an arrangement corresponding to the grid size of a titer plate with 384, 1536 or even higher number of wells. It is precisely this arrangement in the same grid dimension that enables parallel and thus sufficiently rapid transfer of samples from a titer plate to the electrophoresis chip.
  • the application of the samples can therefore be simplified to the extent that pipettes, the volumes of which cannot be reduced arbitrarily, are no longer required. Rather, quantities are sufficient that can be attached to a small structure, for example a thin wire end, by adsorption and released again by diffusion.
  • a metal wire for example a diameter of 50 ⁇ m to 150 ⁇ m, is immersed in the sample, for example in the recess of a microtiter plate, and then introduced into the sample application area in the separation channel of the electrophoresis chip.
  • cathode and anode are used in the description and in the claims instead of the general terms electrode and counter electrode.
  • electrode and counter electrode are used in the description and in the claims instead of the general terms electrode and counter electrode.
  • reverse assignment as anode and cathode is also possible.
  • the invention thus comprises any assignment of the electrodes and counter electrodes.
  • the cathodes and the anode can be part of at least one separate unit which is connected to the electrophoresis chip. According to an advantageous embodiment, however, the cathodes and the anode are integrated in the substrate. This considerably simplifies the construction and handling of the electrophoresis chip in that no additional unit having the electrodes, for example a circuit board with pins as electrodes, has to be arranged and contacted over the electrophoresis chip. By eliminating the sample supply units with the injection channels, the electrodes connected to the sample supply are eliminated. The remaining electrodes required for electrophoretic separation can be integrated on the chip. These can be attached to the top, the bottom or in the chip material itself.
  • the cathodes and the anode are advantageously arranged in such a way that the same electric field prevails in all separation channels.
  • the cathodes and / or the anode are preferably designed as cathode channels or anode channel. These channels are made as grooves in the plate-shaped substrate. The bottoms of these grooves have at least one layer of an electrically conductive material.
  • the intermediate channels connected to the sample application areas open into cathode channels, while the separation channels beginning with the sample application areas open into the anode channel.
  • the configuration of the cathodes and anodes as channels creates a large reservoir volume for buffer solutions in the plane of the electrophoresis chip in comparison with the known solutions. It is therefore not necessary to enlarge the reservoirs beyond the level of the chip, for example by fitting pipette tips filled with buffer solution or a thick elastomer film (P. C. Simpson, op. Cit.) Provided with holes for the buffer reservoir.
  • the layer of an electrically conductive material is advantageously at least one metal layer, in particular a gold layer.
  • the metal possibly with an adhesive layer in between, is evaporated onto the groove bottoms.
  • Such layers thicknesses of 10 nm to 200 nm are advantageous, can be applied inexpensively using coating methods, such as sputtering or vapor deposition. In order to achieve a coating of only the groove bottoms, other areas of the chip are advantageously covered with a mask.
  • the cathode channels are preferably designed to be electrically and fluidly connected. Such a fluidically connected channel structure makes it easier to fill with a buffer solution or a gel and also increases the available reservoir volume.
  • the bottoms of this channel structure are advantageously provided with at least one electrically conductive layer throughout.
  • the cathode channels are advantageously arranged in a column-like manner between columns of sample application areas such that two adjacent cathode channels each surround two columns of sample application areas laterally.
  • the cathode channels arranged in columns in this way are connected to one another above the top row of the sample application areas by a transverse cathode channel.
  • the separation channels advantageously open parallel to one another into an anode channel oriented transversely thereto.
  • This area advantageously serves as a detection area.
  • the electrophoresis chip is designed to be optically transparent at least in this area.
  • a channel is advantageously arranged on the chip as a buffer reservoir parallel to the transverse cathode channel and / or parallel to the anode channel. This is connected to the cathode channel or anode channel at least in a central region.
  • the bottom of the channel serving as a buffer reservoir can also be coated in an electrically conductive manner.
  • the cathode channels, the anode channel and / or the one serving as a buffer reservoir Channel advantageously a width> 200 ⁇ m, particularly advantageously from 500 ⁇ m to 3 mm.
  • These channels thus have a significantly greater width than the separating channels, the width and / or depth of which is advantageously in the range from 0.1 ⁇ m to 200 ⁇ m, particularly advantageously in the range from 0.5 ⁇ m to 100 ⁇ m.
  • the separation channels preferably have the same lengths. This requires a meandering guidance of the channels in curves or loops.
  • the largest possible curve radii advantageously> 200 ⁇ m, in particular between 300 ⁇ m and 700 ⁇ m, should be provided.
  • Each separation channel advantageously has the same number of right and left curves in order to achieve the same separation conditions.
  • Each separation channel extends between the sample application area and the anode.
  • the sample application area is formed by a widened area of the separation channel, this area advantageously having a diameter of 50 ⁇ m to 500 ⁇ m, in particular of 80 ⁇ m to 300 ⁇ m.
  • the depth of the sample application area is advantageously 10 ⁇ m to 300 ⁇ m. A depth that is essentially the same as that of the separation channels is preferred, which results in a significant simplification of the manufacture.
  • the intermediate channel connecting the sample application areas to a cathode or a cathode channel advantageously has a width greater than or equal to the width of the separation channel leading away from the sample application area.
  • the width is also selected to be less than or equal to the diameter of the sample application area. This ensures a good fluidic connection with the cathode channel, with only a slight diffusion of an applied sample into the intermediate channel.
  • Widths of the intermediate channel from 50 ⁇ m to 400 ⁇ m are preferred.
  • the depth is advantageously selected to be equal to the depth of the sample application area, the separation channel and / or the electrode channel. It is conceivable that a Area of the bottom of the intermediate channel has a metallic coating which is connected as an electrode. For this purpose, the metallic coating can extend from the electrode channel into the intermediate channel.
  • the electrophoresis chip has a cover plate which has holes as through openings at least in the area of the sample application areas.
  • the plate-shaped substrate with the channels structured into it is preferably a one-piece polymer part produced by molding.
  • the cover plate is likewise preferably a polymer film which has corresponding holes.
  • Suitable materials are, for example, polymethyl methacrylate (PMMA), polycarbonates (PC), cycloolefinic copolymers (COC), polyoxymethylene (POM), polystyrene (PS), polyurethanes or silicones.
  • PMMA polymethyl methacrylate
  • PC polycarbonates
  • COC cycloolefinic copolymers
  • POM polyoxymethylene
  • PS polystyrene
  • silicones silicones.
  • the use of other materials, such as glass or silicon, for the substrate and / or the cover plate can also be considered.
  • the cover plate can be connected to the substrate using adhesive methods known from microfluidic systems. If both parts are polymer parts, a solvent is suitable for the connection, for example or solvent mixture which has swelling or dissolving properties with respect to these materials.
  • the individual cathodes or the coherent cathode channel structure and the anode are connected to electrical connection contacts which extend through to the top and / or bottom of the electrophoresis chip.
  • the connection contacts are not on the top and / or bottom but on one or more sides of the circumference of the chip.
  • the corresponding device for operating at least one electrophoresis chip has a holding and holding device for fixing the spatial position of the chip. Electrical connection contacts are provided which automatically cause electrical contact when the electrophoresis chip is inserted. An externally shielded electrical power supply is also provided.
  • the device is particularly suitable for operating the electrophoresis chips according to the invention. By separating the electrophoresis chip as a disposable part and the operator unit, it is possible to carry out electrophoretic separations of a large number of samples inexpensively and with little human effort. Automation using laboratory robots is also conceivable.
  • the device is advantageously designed in such a way that at least the detection area of the separation channels is accessible from the top for detection, for example in a fluorescence microscope.
  • the device is advantageously designed so flat that it can be used under a microscope objective.
  • At least part of a system for the detection of optionally marked samples, in particular biomolecules, present in the separation channels of the electrophoresis chip is particularly advantageously integrated into the device.
  • FIG. 2a shows a section of the electrophoresis chip according to FIG. 1 as an SEM image
  • FIG. 2b shows a section of the negative mold for producing the electrophoresis chip according to FIG. 1 as an SEM image
  • FIG. 3 shows the electrophoresis chip according to FIG. 1 cut from the side
  • FIG. 6 shows a device for operating an electrophoresis chip with an inserted chip according to FIG. 1 in a perspective representation cut from the side
  • FIG 7 shows an electropherogram obtained with an electrophoresis chip according to the invention.
  • FIG. 1 shows an electrophoresis chip 1 according to the invention in a top view from above.
  • the chip 1 consists of a thin plate made of a polymeric material as the substrate 2.
  • the structures shown are introduced as depressions in the form of grooves in the top of the plate.
  • the separating channels 3a, 3b, ... and the intermediate channels 8a, 8b, ... are shown as black lines.
  • the 96 sample application areas 7a, 7b, ..., which can be recognized as black dots, are arranged in a matrix in 8 rows and 12 columns. These sample application areas are connected to the cathodes 5a, 5b, ...
  • Each sample application area 7a, 7b, ... is integrated in the beginning of a separation channel 3a, 3b, ..., the sample application area being formed by a widened area of the separation channel 3a, 3b, ...
  • the separation channels 3a, 3b, ... are guided in loops such that they have the same length and the same number of bends. All separating channels 3a, 3b, ... open into the anode 6, the channels being guided parallel to one another in the detection area located in front of the anode.
  • the cathodes 5a, 5b, ... and the anode 6 are integrated in the substrate 2 and are designed in the form of grooves 22a, 22b, ... and 23 as cathode channels 20a, 20b, ... and as anode channel 21, respectively .
  • the groove bottoms 24a, 24b, ... and 25 are coated with an electrically conductive material 26.
  • Two columns S1, S2; S3, S4; ... of sample application areas 7a, 7b, ... are each of two cathode channels 20a, 20b; 20b, 20c; 20c, 20d; ... surrounded on the side.
  • the 7 cathode channels 20a to 20g are connected to one another both electrically and fluidically above the top line of sample application areas by a transverse cathode channel 20h, so that a comb-like cathode structure is present.
  • a channel 27 serving as a buffer reservoir is arranged parallel and above the transverse cathode channel 20h, both being connected to one another in the central region.
  • the anode channel 21 is connected in a central region to a channel 28, which is arranged parallel and below this and serves as a buffer reservoir.
  • PMMA was chosen as the substrate material, the chip 1 having an external dimension of 61 x 62 x 2 mm.
  • the 96 sample application areas 7a, 7b, ... were arranged in the grid dimension of a commercially available 384 microtiter plate, around a parallel one Allow transfer of 96 samples each.
  • the separation channels 3a, 3b, ... of a uniform length of 67 mm each had a width and depth of 50 ⁇ m. Separating channels 3a, 3b, ... running parallel were separated from one another by webs with a width of 50 ⁇ m.
  • the radii of the bends of the separation channels were between 360 ⁇ m and 500 ⁇ m. The number of turns was minimal and chosen the same for all separation channels.
  • Each sample application area 7a, 7b, ... was formed by a widened area located in the beginning of the separation channel and having a diameter of approximately 150 ⁇ m.
  • the intermediate channels 8a, 8b, ... had a width of 100 ⁇ m and a depth of 50 ⁇ m.
  • the electrode channels 20a, 20b, ..., 21 and the channels 27, 28 serving as a buffer reservoir were chosen to be relatively wide with a width of approximately 3 mm.
  • the top of the substrate 2 was covered with a 125 ⁇ m thick PMMA film, not shown here, which had holes in the area of the sample application areas 7a, 7b,.
  • the PMMA substrate 2 was microstructured by embossing.
  • a metallic mold insert obtained by electroplating was used as the stamp.
  • a silicon wafer was first structured as a negative using a chrome mask using photolithography and ASE (Advanced Silicon Etching) techniques. Starting from this negative mold, the nickel mold insert was obtained by double galvanic molding. It is also conceivable to use the silicon wafer directly as an embossing stamp, it being advantageous to mechanically stabilize the silicon wafer for this purpose by bonding on a glass plate. In order to avoid a double galvanic impression, a corresponding silicon structure can first be produced in the positive and this can then be galvanically molded in order to obtain a metallic mold insert as a negative mold.
  • FIG. 2a shows a detail from FIG. 1 as a picture in a scanning electron microscope (SEM), with corresponding parts the same reference numerals are used as in Fig. 1.
  • a separation channel 3d can be seen, which is widened in its beginning to form an approximately circular sample application area 7d. This sample application area is connected to the cathode 5a via the intermediate channel 8d.
  • the routing of the separation channels 3a, 3b, ... can be seen in loops in order to achieve an equal length before the separation channels are guided directly parallel to one another. Due to the representation as an SEM image, all channels, including the cathode channels 20a, 20b, ... are shown in black. Furthermore, due to distortion effects, the cathode channels 20a, 20b, ... appear to be barrel-shaped and not straight, as in FIG. 1.
  • FIG. 2b Another SEM image is shown in Figure 2b.
  • a section of the negative mold in silicon is shown for producing the structures shown in FIG. 2a.
  • the same reference numerals as in FIG. 2a were used as negative for corresponding structures.
  • the section shown includes the sample application area 7d and the leading areas of the intermediate channel 8d and the separation channel 3d in the form of webs. The same height of these three structures and the circular configuration of the sample application area 7d can be clearly seen.
  • FIG. 3 shows a section of the electrophoresis chip 1 shown in FIG. 1 along the line III-III, wherein for the sake of clarity no true-to-scale representation has been chosen. All channels are in the form of grooves 4a to 4p, 8a, 8p, 22a, 22b in the top of the substrate 2.
  • a cover plate 10, not shown in FIG. 1, is applied as a film to the substrate 2.
  • the film 10 In the area of the sample application areas 7a and 7p, the film 10 each has an opening 11a, 11p.
  • the sample application areas 7a and 7p are connected to the cathode channels 20a and 20b via the intermediate channels 8a and 8p.
  • the cathode channels 20a and 20b have an electrically conductive material 26 on their groove bottoms 24a and 24b, which serves as the actual cathode 5a and 5b.
  • the separation channels 3a to 3p which are parallel to each other, are shown.
  • FIG. 4 shows a cover plate 10 in the form of a film with 96 openings 11a, 11b, ... in a perspective view, 8 holes each being arranged in 12 columns S1, S2, ...
  • the film 10 has a passage opening 31a, 31b in the form of a slot above and below the columns S1, S2, ...
  • the position of these slits 31a, 31b corresponds to the position of the channels 27, 28 serving as a buffer reservoir and thus enables the channels to be filled with a buffer or gel.
  • a hole is provided towards the corners of the cover plate 10, which corresponds to structures on the substrate, and thus enables easier adjustment of the two parts to one another.
  • the holes 11a, 11b, ... or the slots 31a, 31b can be obtained by laser ablation, water jet cutting, drilling, milling or punching.
  • FIG. 5 shows a top view of a shadow mask 40 which is used to apply the electrically conductive material 26 to the groove bottoms 24a, 24b, ..., 25 of the electrode channels 20a, 20b, ... 21 and the channels 27 serving as a buffer reservoir. 28 is used.
  • this mask 40 is positioned over the substrate 2 in such a way that the groove bottoms to be coated match the corresponding structures of the mask 40 which are designed as through openings 41.
  • An electrically conductive material is then deposited from the gas phase, for example by vapor deposition. First a 100 nm thick titanium or aluminum layer is evaporated as an adhesion promoter, then a 50 nm thick gold layer is deposited.
  • the shadow mask itself was produced by means of photolithographic processes using ASE (Advanced Silicon Etching) techniques.
  • a device 50 according to the invention for operating an electrophoresis chip 1 is shown in perspective in section in FIG. 6 from the side.
  • the device 50 comprises an upwardly open housing 51, the inner cross section of which is dimensioned such that an electrophoresis chip 1 can be inserted.
  • a circuit board 53 connected to an electrical supply line 55, electrical conductor tracks 54 leading to electrical supply on the circuit board 53 leading to electrical connection contacts 52a, 52b, 52c in the form of sockets.
  • the electrophoresis chip 1 has corresponding electrical connection contacts 30a, 30b, 30c in the form of pins, so that when the chip 1 is inserted, these pins engage in the corresponding sockets of the circuit board 53, and an electrical contact is thus automatically effected.
  • HEC hydroxyethyl cellulose
  • PVP polyvinyl pyrrolidone
  • electrophoresis chip using a sample application area integrated into the beginning of the separation channel, one could use conventional electrophoresis chips comparable separation result can be achieved. In this way, in particular the use of a cross-channel structure as a sample feed unit and the associated application of larger sample quantities can be saved.

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Abstract

Puce d'électrophorèse et appareil permettant de faire fonctionner ladite puce. Dans les puces d'électrophorèse connues, le nombre relativement élevé des réservoirs à remplir et à mettre en contact ne permet pas une intégration des électrodes. La présente invention permet de réaliser une puce d'électrophorèse (1) dont les zones d'application d'échantillons sont placées dans la grille de plaquettes de microtitrage, qui ne présente pas ces inconvénients. A cet effet, une zone (7a, 7b, ...) d'application d'échantillons est précisément intégrée au début de chaque canal de séparation, ladite zone étant constituée par un élargissement du canal de séparation (3a, 3b, ...) et reliée à une cathode (5a, 5b, ...) via un canal intermédiaire (8a, 8b, ...). La présente configuration permet une intégration des électrodes (5a, 5b, ..., 6) sous forme de canaux électrodes (20a, 20b, ..., 21), ainsi qu'une application parallèle des échantillons.
PCT/EP2000/002510 1999-03-30 2000-03-22 Puce d'electrophorese et appareil permettant de faire fonctionner ladite puce WO2000058721A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU39633/00A AU3963300A (en) 1999-03-30 2000-03-21 Electrophoresis chip and device for operating an electrophoresis chip
DE10080771T DE10080771D2 (de) 1999-03-30 2000-03-22 Elektrophoresechip sowie Gerät zum Betreiben eines Elektrophoresechips

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19914354 1999-03-30
DE19914354.4 1999-03-30

Publications (1)

Publication Number Publication Date
WO2000058721A1 true WO2000058721A1 (fr) 2000-10-05

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Country Status (3)

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AU (1) AU3963300A (fr)
DE (1) DE10080771D2 (fr)
WO (1) WO2000058721A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003058227A1 (fr) * 2001-11-30 2003-07-17 Capital Biochip Company, Ltd. Appareil a puce d'electrophorese capillaire pour la detection des polymorphismes de nucleotides et de mononucleotides
EP1407257A1 (fr) * 2001-06-20 2004-04-14 Cytonome, Inc. Dispositif de separation microfabrique utilisant une paroi virtuelle afin d'interfacer des fluides
US8715558B2 (en) 2010-05-03 2014-05-06 Indian Institute Of Technology Bombay Capillary electrophoresis chips

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998055852A1 (fr) * 1997-06-06 1998-12-10 Caliper Technologies Corp. Structures microfabriquees facilitant l'introduction de fluides dans des dispositifs microfluidiques
WO1999010735A1 (fr) * 1997-08-28 1999-03-04 Caliper Technologies Corporation Interfaces ameliorees de detecteur/controleur pour systemes microfluidiques
WO1999014368A2 (fr) * 1997-09-15 1999-03-25 Whitehead Institute For Biomedical Research Procede et appareil pour le traitement d'un echantillon d'analyte biomoleculaire au moyen d'un dispositif microfabrique

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
WO1998055852A1 (fr) * 1997-06-06 1998-12-10 Caliper Technologies Corp. Structures microfabriquees facilitant l'introduction de fluides dans des dispositifs microfluidiques
WO1999010735A1 (fr) * 1997-08-28 1999-03-04 Caliper Technologies Corporation Interfaces ameliorees de detecteur/controleur pour systemes microfluidiques
WO1999014368A2 (fr) * 1997-09-15 1999-03-25 Whitehead Institute For Biomedical Research Procede et appareil pour le traitement d'un echantillon d'analyte biomoleculaire au moyen d'un dispositif microfabrique

Non-Patent Citations (2)

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Title
SIMPSON P C ET AL: "HIGH-THROUGHPUT GENETIC ANALYSIS USING MICROFABRICATED 96-SAMPLE CAPILLARY ARRAY ELECTROPHORESIS MICROPLATES", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA,US,NATIONAL ACADEMY OF SCIENCE. WASHINGTON, vol. 95, March 1998 (1998-03-01), pages 2256 - 2261, XP000901185, ISSN: 0027-8424 *
WOOLLEY A T ET AL: "HIGH-SPEED DNA GENOTYPING USING MICROFABRICATED CAPILLARY ARRAY ELECTROPHORESIS CHIPS", ANALYTICAL CHEMISTRY,US,AMERICAN CHEMICAL SOCIETY. COLUMBUS, vol. 69, no. 11, 1 June 1997 (1997-06-01), pages 2181 - 2186, XP000696550, ISSN: 0003-2700 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1407257A1 (fr) * 2001-06-20 2004-04-14 Cytonome, Inc. Dispositif de separation microfabrique utilisant une paroi virtuelle afin d'interfacer des fluides
EP1407257A4 (fr) * 2001-06-20 2004-11-24 Cytonome Inc Dispositif de separation microfabrique utilisant une paroi virtuelle afin d'interfacer des fluides
WO2003058227A1 (fr) * 2001-11-30 2003-07-17 Capital Biochip Company, Ltd. Appareil a puce d'electrophorese capillaire pour la detection des polymorphismes de nucleotides et de mononucleotides
US7527719B2 (en) 2001-11-30 2009-05-05 Capitalbio Corporation Capillary electrophoresis chip apparatus for detecting nucleotide polymorphism and single nucleotide polymorphism
US8715558B2 (en) 2010-05-03 2014-05-06 Indian Institute Of Technology Bombay Capillary electrophoresis chips

Also Published As

Publication number Publication date
DE10080771D2 (de) 2003-04-17
AU3963300A (en) 2000-10-16

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