EP1606671A2 - Verfahren und vorrichtung zum benetzen eines substrats mit einer flüssigkeit - Google Patents
Verfahren und vorrichtung zum benetzen eines substrats mit einer flüssigkeitInfo
- Publication number
- EP1606671A2 EP1606671A2 EP04722244A EP04722244A EP1606671A2 EP 1606671 A2 EP1606671 A2 EP 1606671A2 EP 04722244 A EP04722244 A EP 04722244A EP 04722244 A EP04722244 A EP 04722244A EP 1606671 A2 EP1606671 A2 EP 1606671A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- wetting
- substrate
- protective layer
- wetted
- liquid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
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Classifications
-
- G—PHYSICS
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- G03F—PHOTOMECHANICAL PRODUCTION OF TEXTURED OR PATTERNED SURFACES, e.g. FOR PRINTING, FOR PROCESSING OF SEMICONDUCTOR DEVICES; MATERIALS THEREFOR; ORIGINALS THEREFOR; APPARATUS SPECIALLY ADAPTED THEREFOR
- G03F7/00—Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor
- G03F7/26—Processing photosensitive materials; Apparatus therefor
- G03F7/40—Treatment after imagewise removal, e.g. baking
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
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- 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
- B01L3/0289—Apparatus for withdrawing or distributing predetermined quantities of fluid
- B01L3/0293—Apparatus for withdrawing or distributing predetermined quantities of fluid for liquids
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
-
- C—CHEMISTRY; METALLURGY
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
- C40B50/14—Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
-
- G—PHYSICS
- G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
- G03F—PHOTOMECHANICAL PRODUCTION OF TEXTURED OR PATTERNED SURFACES, e.g. FOR PRINTING, FOR PROCESSING OF SEMICONDUCTOR DEVICES; MATERIALS THEREFOR; ORIGINALS THEREFOR; APPARATUS SPECIALLY ADAPTED THEREFOR
- G03F1/00—Originals for photomechanical production of textured or patterned surfaces, e.g., masks, photo-masks, reticles; Mask blanks or pellicles therefor; Containers specially adapted therefor; Preparation thereof
- G03F1/68—Preparation processes not covered by groups G03F1/20 - G03F1/50
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00277—Apparatus
- B01J2219/00351—Means for dispensing and evacuation of reagents
- B01J2219/00364—Pipettes
- B01J2219/00367—Pipettes capillary
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
- B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
- B01J2219/00277—Apparatus
- B01J2219/00351—Means for dispensing and evacuation of reagents
- B01J2219/00373—Hollow needles
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Definitions
- the invention relates to a method and a device for wetting a substrate with a liquid, and to a liquid-wetted substrate that can be produced with the method according to the invention.
- wetting a substrate with a liquid is widely used in industry and science. Especially in the field of microstructuring of surfaces for the life sciences, medical technology and sensor technology, in addition to the classic lithographic methods, wetting processes have also become increasingly important in recent years.
- wetting methods for the lateral structuring of surfaces can be roughly divided into two classes: methods with direct contact of the wetting device with the substrate, and methods without direct contact.
- micro-contact printing In the structuring methods with direct contact, the micro-contact printing ⁇ CP (mico-contact printing) should be emphasized, which was first published by Whitesides 1994 (A. Kumar, GM Whitesides, Science, 1994, 263, 60; US-A-6 048 623) was presented.
- a micro-structured stamp is wetted with a liquid, then brought into contact with the substrate to be processed and thus the surface brought to the substrate to be processed and thus imprinted a lateral chemical structure on the surface.
- a major difficulty of this technique is the realization of a uniform contact between the stamp and the substrate, which is of crucial importance for the success or the quality.
- the ink-jet printing methods should be mentioned here by way of example.
- the liquid is absorbed in the printhead and positioned over the desired location on the substrate.
- a force is exerted on the liquid by a piezoelectric crystal or a pump, so that a drop leaves the contact head and is transferred to the substrate.
- the size of the wetted area is also determined by the surface energies of the materials involved.
- the equilibrium state of the drop defined by the contact angle between liquid and substrate, is highly dependent on factors such as Surface roughness, chemical inhomogeneities of the material, variations in the surrounding atmosphere and of course impurities.
- the transferred drops are wetted on a macroscopic substrate in very different ways. The processes from the prior art are therefore subject to fundamental limits with regard to tolerances in spot sizes and wetting volumes.
- the object of the present invention is therefore to create a method and a device for wetting substrates with a liquid which do not have the disadvantages of the prior art.
- Liquid not only pure liquid substances, but also liquids with de- tergenz, any kind of dissolved organic or inorganic substances, as well as emulsions, suspensions and colloidal solutions.
- Laser ablation partial or complete removal of organic or inorganic protective layers, but also the removal of contaminants on a substrate by exposure to laser light.
- Solder masking lacquer known from printed circuit board technology, which is applied to printed circuit boards to prevent the formation of solder bridges during automated soldering.
- Protective layer applied to the substrate to be processed before the actual wetting Any material can be used for this, which forms a closed layer on the substrate surface and thus separates it from the environment and can be removed at a later time by laser ablation partially and without residue.
- This protective layer can consist of both organic and inorganic materials and, depending on the type of substrate and the application requirements, can be physisorbed, chemisorbed or covalently bound and applied using any technique.
- Solid substrates include both plastics and metals, semiconductors, glasses, composites or porous materials.
- the term surface is independent of the spatial dimensions of the surface and also includes nanoparticles (particles or clusters of a few individual to several hundred thousand surface atoms or molecules).
- PNA Peptide nucleic acid synthetic DNA or RNA in which the
- N (COCH 2 base) -CH 2 CO unit hybridizes PNA with DNA).
- Nucleic acid at least two covalently linked nucleotides or at least two covalently linked pyrimidine (e.g. cytosine, thymine or uracil) or purine bases (e.g. adenine or guanine).
- the term nucleic acid refers to any "backbone" of the covalently linked pyrimidine or purine bases, such as. B. on the sugar-phosphate backbone of DNA, cDNA or RNA, on a peptide backbone of the PNA or on analog structures (e.g. phosphoramide, thio-phosphate or dithio-phosphate backbone).
- An essential feature of a nucleic acid in the sense of the present invention is that it can bind naturally occurring cDNA or RNA in a sequence-specific manner.
- Nucleic acid - Nucleic acid of unspecified base length e.g. nuc-oligomer, linseic acid octamer: a nucleic acid with any backbone in which 8 pyrimidine or purine bases are covalently bound to one another.
- Oligomer equivalent to nucleic acid oligomer Oligonucleotide equivalent to oligomer or nucleic acid oligomer, e.g. B. a DNA, PNA or RNA fragment unspecified base length.
- Oligo Abbreviation for oligonucleotide ss Single Strand
- alkyl denotes a saturated hydrocarbon radical which is straight-chain or branched (for example ethyl, isopropyl or 2,5-dimethylhexyl etc.).
- alkyl refers to a group with two available valences for covalent linkage (e.g. - CH 2 CH 2 -, -CH 2 CH 2 CH 2 - or -CH 2 C (CH 3 ) 2 CH 2 CH 2 C (CH 3 ) 2 CH 2 - etc.).
- Alkenyl alkyl groups in which one or more of the C-C single bonds are replaced by C C double bonds.
- Alkynyl alkyl or alkenyl groups in which one or more of the C-C single or C C double bonds are replaced by C ⁇ C triple bonds.
- C18 octadecanethiol fluorophore chemical compound that is able to emit a longer-wave (red-shifted) fluorescent light when excited with light.
- Fluorophores fluorescent dyes
- UV ultraviolet
- VIS visible
- IR infrared
- the absorption and emission maxima are typically shifted from each other by 15 to 40 nm (Stokes shift).
- Ligand Term for molecules that are specifically bound by the ligate examples include substrates, cofactors or coenzymes of a protein (enzyme), antibodies (as ligand of an antigen), antigens (as ligand of an antibody), receptors (as ligand of a hormone), hormones (as ligand of a receptor) ) or nucleic acid oligomers (as ligand of the complementary nucleic acid oligomer.
- Ligate Term for (macro) molecule with specific recognition and binding sites for the formation of a complex with a ligand (template).
- Fluorescein resorcin phthalein R any unspecified organic radical as a substituent or side chain.
- Thiol molecules of the general structure HS -Spacer-R or [SSpacer-R] 2 spacers Any molecular connection between two molecules or between a surface atom, surface molecule or a surface molecule group and another molecule, usually alkyl, alkenyl, alkynyl, Heteroalkyl, heteroalkenyl, heteroalkynyl chains.
- Preferred spacers are those of chain length 1-20, in particular chain length 1-14, the chain length being the shortest continuous connection between the structures to be connected.
- the terminal phosphate group of the oligonucleotide is esterified at the 3 'end with (HO- (CH 2 ) 2 -S) 2 to PO- (CH 2 ) 2 -SS- (CH 2 ) 2 -OH, the SS bond being cleaved homolytically and each causes an Au-SR bond.
- the probe oligonucleotide carries a covalently attached fluorophore fluorescein.
- Oligo-Spacer-SS- two identical or different nucleic acid oligomers which are connected to each other via a disulfide bridge, the disulfide bridge being connected to the nucleic acid oligomers via any two spacers and the two spacers having a different chain length (shortest continuous connection between the disulfide bridge and the respective nucleic acid oligomer), in particular any chain length between 1 and 14, and these spacers in turn are connected to various naturally existing on the nucleic acid oligomer. ne or attached to these by modification attached reactive groups.
- These spacers can in turn be bound to various reactive groups that are naturally present on the nucleic acid oligomer or are attached to it by modification and “n” is any integer, in particular a number between 1 and 20.
- n x R-S-S-nucleic acid oligomer to which n disulfide functions are connected via spacers in each case -oligo a spacer, any residue R saturating the disulfide function.
- the spacer for connecting the disulfide function to the nucleic acid oligomer can each have a different chain length (shortest continuous connection between disulfide function and nucleic acid oligomer), in particular any chain length between 1 and 14. These spacers can in turn be attached to different ones of course on the nucleic acid -Oligomer existing or attached to this by modification attached reactive groups.
- the placeholder "n" is any integer, in particular a number between 1 and 20.
- a method for wetting a substrate with a liquid comprises the following method steps: a) providing a substrate with a surface to be wetted; b) providing a wetting liquid; c) applying a protective layer to the substrate, which is the one to be wetted
- the procedure according to the invention largely prevents contamination of the wetting areas and the wear on the
- wetting device minimized.
- structuring of the protective layer allows the areas to be wetted on the substrate to be specified in a simple manner.
- a solid body made of plastic, metal, semiconductor, glass, composite, porous material or a combination of these materials is advantageously provided as the substrate.
- a solid is preferably provided as the substrate, the surface to be wetted of which is formed by a silicon, platinum or gold layer or an oxidic layer or a glass.
- the spatial design of the substrate is not restricted according to the invention. Rather, a macroscopic solid-state disk, a micro- or nanoparticle, for example, can be provided as the substrate.
- the term “wetting liquid” includes in particular purely liquid substances, solutions of organic and / or inorganic substances, emulsions, suspensions or colloidal solutions.
- the material of the protective layer is expediently matched to the substrate material such that the protective layer material is physisorbed, chemisorbed or covalently, coordinatively or via complex formation on the substrate surface to be wetted.
- a protective layer a positive or negative photoresist can be applied to the substrate, preferably sprayed on or spun on.
- a solder resist can also be used as a protective layer for the substrate. It is preferred that the solder resist is applied by screen printing, curtain casting or a spray process.
- an organic polymer in particular made of cellulose, dextran or collagen, is applied to the substrate as a protective layer.
- the organic polymer is preferably spin-coated or physisorbed.
- a self-assembled monolayer made of organic molecules is applied as a protective layer. This is produced in particular by dissolving the organic molecules in an aqueous or organic solvent and bringing the solution into contact with the substrate.
- a particularly preferred embodiment results if a solid is advantageously provided as the substrate, the surface to be wetted is formed by a gold layer and if a self-assembled monolayer of thiols, in particular of the general structure HS-Spacer-R or [S -Spacer-R] 2 is applied.
- R represents any head group and the spacer has a chain length of 1-20, in particular 1-14.
- a solid body is provided as the substrate, the surface to be wetted of which is formed by a silicon or platinum layer, and if as a protective layer a self-assembled monolayer of amines, in particular the general structure H 2 N-Spacer-R is applied.
- R represents any head group and the spacer has a chain length of 1-20, in particular 1-14.
- a solid body is provided as the substrate, the surface to be wetted of which is formed by an oxidic surface or a glass.
- the head group R is expediently selected from the group CH 3 , OH, CO 2 H, NH 2 , NH 3 + or SO 3 " .
- the protective layer is advantageously applied in step c) in the form of a closed layer to the substrate surfaces to be wetted. It can be applied over the entire surface of the entire surface of the substrate or can only cover partial areas of the surface. The protective layer is then expediently removed without residue in the area of the desired wetting areas.
- the protective layer is structured by means of laser ablation, in particular by irradiating partial areas of the protective layer with continuous or pulsed laser radiation of a predetermined wavelength.
- the protective layer is in particular exposed directly to the laser radiation via an optic or a mask in order to expose the wetting areas.
- the surface of the substrate to be wetted is exposed in the area of the wetting areas by the laser radiation. is melted. This results in a reduced surface roughness and an improved homogeneity of the surface of the substrate. In addition, less gold layers are removed from the surface by ablation.
- the wetting areas are advantageously produced with a characteristic extent of approximately 5 ⁇ m to approximately 200 ⁇ m, preferably of approximately 10 ⁇ m to approximately 100 ⁇ m. A value of approximately 20 ⁇ m to approximately 500 ⁇ m, preferably of approximately 50 ⁇ m to approximately 200 ⁇ m, is set as the lateral distance.
- the wetting areas advantageously have an essentially rectangular, elliptical or circular outline.
- additional feed channels are introduced into the protective layer in step d) in order to enable the supply of an analyte liquid to the exposed wetting areas.
- the supply channels are expediently introduced into the protective layer at a depth of 10% to 99%, preferably from 20% to 95%, particularly preferably from 50% to 95% of the thickness of the protective layer.
- the exposed wetting areas are advantageously arranged within the supply channels.
- the wetting device comprises in particular a single needle, capillary, tweezers, a single ring or stamp.
- it can also comprise an arrangement of several needles, capillaries, tweezers, rings, stamps, or an arrangement of various of these elements.
- the wetting device has a liquid-releasing end surface, the lateral extent of which in at least one spatial direction is greater than the lateral extent of the loading areas of networking in this spatial direction. With correct alignment, direct contact between the wetting device and the surface of the substrate can thereby be avoided.
- the end face of the wetting device advantageously has a greater lateral extent in both spatial directions than the wetting areas, so that direct contact between the wetting device and the wetting areas is avoided in all relative orientations.
- the end face of the wetting device is preferably brought into contact with the protective layer adjacent to the wetting area.
- a drop of the wetting liquid can thus be introduced into the structured recess in the protective layer in a controlled manner without direct contact with the substrate surface.
- the end face of the wetting device for applying the wetting liquid can be brought completely over the wetting area and from above into contact with the surface of the protective layer adjacent to the wetting area.
- the end face of the wetting device can be positioned laterally with an accuracy ( ⁇ x, ⁇ y) over a structured protective layer, and the wetting areas are generated with a characteristic lateral extent (x spot , y spo t), which by at least the positioning accuracy ( ⁇ x, ⁇ y) is smaller than the lateral extent (xtip, y «p) of the end face of the wetting device. This ensures that the delivery of a drop is controlled and only takes place over the protective layer.
- modified nucleic acid oligomers are applied in aqueous solution as a wetting liquid.
- the nucleic acid oligomers are one or more modified reactive groups, at least one reactive group being designed for a direct reaction with the surface of the substrate to be wetted.
- the nucleic acid oligomers can also be modified with a fluorophore for subsequent visualization.
- the invention also includes an apparatus for performing the described method.
- a device particularly advantageously contains a wetting device whose end face can be positioned laterally over a structured protective layer with a positioning accuracy of less than 50 ⁇ m, preferably less than 10 ⁇ m.
- the invention also includes a liquid-wetted substrate that can be produced by a previously described method.
- the invention comprises a method for the controlled wetting of structured substrates with a liquid by means of a wetting device consisting of a single needle, capillary, tweezers, a ring or stamp or an arrangement of needles, capillaries, tweezers, rings or stamps ,
- these wetting devices can have tips with any lateral extent, that is to say also and even preferably larger than the lateral area of the laser-ablated, free substrate locations.
- the wetting device of the invention does not require direct contact with the substrate and can therefore be referred to as a pseudo-contact method.
- the substrates are provided with a protective layer in order to to bridge the critical period between the manufacture of the substrate and the wetting of its surface. During this period, the protective layer prevents the accumulation of undesired impurities on the substrate surface.
- any material can be used for the protective layer which forms a closed layer on a surface and thus separates the substrate surface from the surroundings and can be removed at a later point in time without residue, for example by laser ablation.
- an adapted protective layer is advantageously chosen for a given substrate, which is optimized with regard to the adhesion between the substrate and protective layer.
- the protective layer can also be optimized with regard to the liquid to be used. In the case of aqueous solutions, a hydrophilic layer material lends itself so that the liquids wet the feed channels of the invention and air bubbles are avoided. In the case of oily liquids, however, hydrophobic material is preferred.
- organic polymers such as cellulose, dextran or collagen or self-assembled monoagens made from organic molecules such as silanes or thiols are also suitable. It is also conceivable to use lacquers whose special constituents form advantageous functionalizations for special applications when the material dries on the surface.
- the protective layer can be applied, for example, by spraying in the case of the photoresists, by spin coating or physisorption in the case of the organic polymers, or by screen printing or curtain casting in the case of the solder resist Substrate are applied.
- monolayers of organic molecules such as thiols or silanes with variable chain length are applied to the substrate in a self-assembling process.
- the organic molecules are dissolved in aqueous or organic solvents and the solution is brought into contact with the substrate to be coated.
- the deposition process ends in a monolayer of covalently bound molecules on the substrate.
- the organic molecules can also be provided with amine groups (H 2 N-Spacer-R) instead of the thiol groups (SH-Spacer-R), which are then chemically or physisorbed onto platinum or silicon Allow surfaces to attach.
- amine groups H 2 N-Spacer-R
- SH-Spacer-R thiol groups
- protective layers are applied to the substrates from solder resists known from printed circuit board technology.
- solder resists Two-component or one-component solder resists are suitable, which are applied by curtain casting, screen printing or spraying and then in air or by UV radiation can harden.
- An advantage of this process variant is that the thickness of the solder resist layer can be freely adjusted in a wide range, for example in the curtain casting process, by the speed of the substrates under the lacquer curtain.
- laser ablation means not only the partial or complete removal of organic or inorganic protective layers, but also the removal of impurities on a substrate by irradiation with laser light. Laser ablation is advantageously used to remove or structure the applied protective layer at desired locations on the substrate in any geometry. This makes it possible to realize different, precisely defined free substrate areas or areas with a tapered protective layer in different sizes on one and the same substrate design only by changing the laser exposure.
- Another aspect of the solution according to the invention is the melting of the substrate surface with complete removal of the protective layer by means of laser ablation, which can be achieved by adjusting the laser intensity or the irradiation time to the conditions of the substrate and the protective layer.
- this short-term melting of the substrate surface close to the surface also closes pores in the material and thus improves the homogeneity of the free substrate surface.
- ablation removes fewer layers of gold from the surface.
- Laser ablation can be carried out by direct irradiation of the light or by irradiation of the light via an optic or a mask.
- the size or shape of each exposed or structured wetting areas and their lateral distance are arbitrary and only depend on the respective application.
- the wavelength of the laser light used, as well as the irradiation time and the number and duration of the pulses depend on the combination of protective layer and the material of the substrate surface and are preferably optimized for each pair.
- structures from channels and free wetting areas are written into a solder mask using an excimer laser over a plurality of masks in a plurality of process steps, which structures, in addition to the controlled wetting at the free substrate locations by means of the pseudo-contact printing process described, also the targeted one Allow contacting of points connected to one another via channels with a liquid containing an analyte.
- the wetting liquid is in particular with the aid of a needle, capillary, tweezers, a ring or stamp or an arrangement application of needles, capillaries, tweezers, rings or stamps onto the structured substrate.
- the term “pseudo-contact printing” is used for the wetting process in order to differentiate the technology from the known standard method of “contact printing” and to make it clear that, because of the protective layer present and the lateral extent of the tips of the wetting device, which is preferably larger than the free surfaces to be wetted, there is no direct contact between the wetting device and the substrate surface. Since, in addition, the free substrate surface to be wetted is limited by the protective layer of a predetermined height, the wetting device encounters a geometrical barrier of a defined dimension, so that controlled wetting takes place.
- ligates In the context of the invention, both pure liquid substances and any type of dissolved organic or inorganic substances, as well as emulsions, suspensions and colloidal solutions can be used. Possible materials in the sense of the invention are dissolved color pigments or any functionalized polymers and nanoparticles. In the field of sensor technology, all types of ligates can be applied to the substrate with the present invention. Molecules that specifically interact with a ligand to form a complex are referred to as ligates.
- ligates in the sense of the present document are substrates, cofactors or coenzymes as complex binding partner of a protein (enzyme), antibodies (as complex binding partner of an antigen), antigens (as complex binding partner of an antibody), receptors (as complex binding partner of a hormone), hormones (as complex binding partner a receptor), nucleic acid oligomers (as complex binding partner of the complementary nucleic acid oligomer) or metal complexes.
- the free substrate sites are wetted with modified nucleic acid oligomers in aqueous solution.
- the nucleic acid oligomer to be applied to the free surface is modified with one or more reactive groups via a covalently attached spacer of any composition and chain length, these reactive groups preferably being located near one end of the nucleic acid oligomer.
- the reactive groups are groups that can react directly with the unmodified surface.
- HS spacer thiol
- SS disulfide
- the dispenser of the wetting device with any lateral extent (x t j P , y tip ) is positioned with an accuracy of ( ⁇ x, ⁇ y) over the structured protective film and lowered for wetting so that the contact of the Wetting device when dispensing the drop takes place only over the protective layer. This is ensured in particular if the wetting areas have a characteristic lateral extent (x spot . Yspot) which is at least the positioning accuracy less than the extent of the dispenser, that is to say the conditions
- a method for wetting a substrate with a liquid according to an embodiment of the invention is described below in particular with reference to FIG. 1.
- a substrate 10 is provided with a surface 12 to be wetted, FIG. 1 (a).
- the substrate 10 consists of a glass slide with a vapor-deposited, 5 nm thick CrNi contact layer and a gold layer vapor-deposited thereon with a thickness of approximately 200 nm.
- the substrate 10 is incubated at room temperature for 5-12 hours with 1 nmol / l octadecanethiol (C-18; Fluka) in ethanol and, after the incubation, rinsed with ethanol in order to add unbound thiol remove, Fig. 1 (b).
- C-18 1 nmol / l octadecanethiol
- the C18 protective film 14 is patterned by laser ablation to form a plurality of wetting areas 16, as illustrated in FIG. 1 (c).
- the structuring of the C18 protective film is carried out with radiation 18 having a wavelength of 193 nm of an excimer laser 20 from Lambda-Physics.
- the thiols of the protective layer 14 in the wetting areas 16 can be removed without residue.
- the laser bombardment of the substrate 10 also leads to a melting of the gold surface, whereby pores are closed, the roughness is reduced and impurities are removed (FIG. 3).
- the laser radiation is imaged on the substrate in a reduced form via a mask (not shown), which in the exemplary embodiment provides illumination spots with a diameter of 40-100 ⁇ m.
- the wetting areas are burned into the protective layer with a lateral distance of, for example, 200 ⁇ m.
- FIG. 2 shows SEM images of wetting areas 16 exposed by laser ablation in a protective layer 14.
- a stop mask protective layer was used instead of the C18 protective layer of FIG. 1 for these SEM images.
- a two-component solder resist (Elmer GL 2467 SM-DG, from Peters) is applied to the substrate in a curtain casting process known from printed circuit board technology in order to form a protective layer for the surface of the substrate.
- Transport speed of the substrate 10 under the paint curtain can achieve any thickness of the protective layer in the range of approximately 10-150 ⁇ m.
- the protective layer is structured with an excimer laser from Lambda-Physics by laser ablation.
- 90-150 pulses of 20 ns with an area performance of 600-1200 mJ / cm 2 remove the varnish without residues and ensure that the gold substrates melt close to the surface, which closes the existing pores Roughness reduced and surface contamination removed.
- the laser can be imaged on the substrate in a reduced size using various masks, the area intensity of the irradiation being set using the imaging device. Depending on the mask, different geometries of the ablated regions can be realized. , , ,
- FIG. 2 illustrates that both rectangular or square cross sections (FIG. 2 (a)) and round cross sections, as shown in FIG. 2 (b), are possible.
- FIG. 3 shows in (a) an AFM image of a gold surface which was melted in a circular partial area by laser bombardment, and in FIG. 3 (b) an elevation profile along the line B-B of FIG. 3 (a). It can clearly be seen that the melting reduces the roughness of the surface and increases the homogeneity of the irradiated surface. This facilitates the connection of probe molecules to the wetting regions 16 described below.
- FIG. 1 (d) shows the wetting of the structured substrates by means of a wetting device 22 with nucleic acid oligomers in the pseudo-contact printing process.
- the oligonucleotides are synthesized in an automatic oligonucleotide synthesizer (Expedite 8909; ABI 384 DNA / RNA synthesizer) according to the synthesis protocols recommended by the manufacturer for a 1.0 ⁇ mol synthesis.
- the oxidation steps are carried out with a 0.02 molar iodine solution in order to avoid oxidative cleavage of the disulfide bridge.
- Modifications to the 5 ' position of the oligonucleotides are carried out with a coupling step which is extended to 5 min.
- the amino modifier C2 dT (Glen Research 10-1037) is built into the sequences with the respective standard protocol. The coupling efficiencies are determined online during the synthesis via the DMT cation concentration photometrically or conductometrically.
- the oligonucleotides are deprotected with concentrated ammonia (30%) at 37 ° C for 16 h.
- the oligonucleotides are purified using RP-HPL chromatography according to standard protocols (eluent: 0.1 molar triethylammonium acetate buffer, acetonitrile), and the characterization is carried out using MALDI-TOF MS.
- the amine-modified oligonucleotides are coupled to the corresponding activated fluorophores (eg fluorescein isothiocyanate) in accordance with the conditions known to the person skilled in the art. The coupling can take place both before and after the oligonucleotides have been bound to the surface.
- buffer phosphate buffer, 0.5 molar in water, pH 7
- the free propanethiol present in the incubation solution is also co-adsorbed by forming an Au-S bond (incubation step).
- this single strand can also be hybridized with its complementary strand.
- Split pin needles 22 (Arraylt chipmarker pins from Tele-Chem) are used for the assignment with the spotter from Cartesian Technologies (MicroSys PA), which have a loading volume 24 of 0.2 to 0.6 ⁇ L and volumes 26 of Dispense approximately 1 nL per wetting process.
- a side view of the needle 22 during the wetting process and a wetted wetting area 16 is shown in FIG. 1 (e).
- the contact surface 28 of the needles 22 has a diameter of approximately 130 ⁇ m and is thus significantly larger than the wetting areas 16 of the substrate that are exposed during laser ablation.
- the needle is positioned over the substrate with an accuracy of 10 ⁇ m at a humidity of around 70-80%.
- the drop 26 is released when the tip comes into contact with the protective layer 14 and there is no direct contact of the needle 22 with the surface 12 of the substrate 10 to be wetted. This situation is shown in the left partial image of FIG. 1 (e). After wetting has taken place, a drop of liquid 30 is applied in a controlled manner to the wetting point 16 of the substrate (right partial image of FIG. 1 (e)).
- a fluorescence intensity measurement on the Au-ss-oligo-fluorescein system will now be described as an application example.
- wetting areas 16 are also formed on a structured substrate 10 Functionalized nucleic acid oligomers.
- a modified oligonucleotide of the sequence 5'-fluorescein-AGC GGA TAA CAC AGT CAC CT-3 '[C 3 -SSC 3 -OH] is immobilized on gold (50 ⁇ mol oligonucleotide in phosphate buffer (K ⁇ PO ⁇ KH ⁇ O . ⁇ 500 mmolar, pH 7), subsequent coating with propanethiol 1 mM in water) and in the form Au-S (CH 2 ) 2-ss-oligo-Fluorescein the fluorescence intensity of the surface with a fluorescence scanner from Lavision Biotech certainly.
- 150 ⁇ l of the medium are placed on the gold surface and then covered with
- FIG. 4 shows the fluctuations in the fluorescence intensity of several identical measurement spots.
- the serial number of the measurement spots is plotted on the abscissa, the fluorescence intensity measured in any units on the ordinate.
- the nucleic acid in the measured values of FIG. 4 (a), the nucleic acid
- a solder resist is used as a protective layer and is structured with leads for liquid analytes in order to produce wetting areas.
- supply channels for liquids can also be written in thick solder mask layers (for example 100-150 ⁇ m).
- a first structuring step different types of channels are cut into the lacquer via a first mask, the depth of these channels being adjustable by the number of pulses.
- a channel depth of about 80 - 120 ⁇ m is achieved with about 540 - 900 pulses (20 ns) of the laser with an area performance of 600 - 1200 mJ / cm 2 .
- the remaining lacquer is then removed via a second mask in individual areas within the channels of the first structuring step by additional laser exposure with approximately 90-150 pulses (20 ns), and the substrate is thus exposed and melted. These exposed substrate sites are now wetted with nucleic acid oligomers as described above.
- analyte such as Liquids which contain potentially complementary nucleic acid oligomers are brought into contact, and thus the analyte liquid required for an analysis is significantly reduced.
- a channel structure that e.g. only one part of the exposed substrate locations per channel connects an arrangement of n linear channels, each of which contains all m wetting areas of a column of a uniform spot matrix of the dimension n x m, expediently 10 ⁇ n, m ⁇ 1000.
- Another channel structure that connects all of the exposed substrate sites to one another is a single channel that meanders all of the exposed substrate sites of the uniform wetting area matrix of the dimension n x m, expediently 10 ⁇ n, m 1000 1000.
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Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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DE2003112628 DE10312628A1 (de) | 2003-03-21 | 2003-03-21 | Verfahren und Vorrichtung zum Benetzen eines Substrats mit einer Flüssigkeit |
DE10312628 | 2003-03-21 | ||
PCT/EP2004/002977 WO2004082814A2 (de) | 2003-03-21 | 2004-03-22 | Verfahren und vorrichtung zum benetzen eines substrats mit einer flüssigkeit |
Publications (1)
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EP1606671A2 true EP1606671A2 (de) | 2005-12-21 |
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EP04722244A Withdrawn EP1606671A2 (de) | 2003-03-21 | 2004-03-22 | Verfahren und vorrichtung zum benetzen eines substrats mit einer flüssigkeit |
Country Status (5)
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US (1) | US20060257630A1 (ja) |
EP (1) | EP1606671A2 (ja) |
JP (1) | JP2006521538A (ja) |
DE (1) | DE10312628A1 (ja) |
WO (1) | WO2004082814A2 (ja) |
Families Citing this family (8)
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DE102007044664B4 (de) | 2007-09-18 | 2012-01-05 | Friz Biochem Gesellschaft Für Bioanalytik Mbh | Verdrängungsassay zur Detektion von Nukleinsäureoligomer-Hybridisierungsereignissen |
GB2455746B (en) * | 2007-12-19 | 2011-03-09 | Cambridge Display Tech Ltd | Electronic devices and methods of making the same using solution processing techniques |
DE102008048342B4 (de) * | 2008-09-22 | 2013-01-24 | Laser-Laboratorium Göttingen eV | SERS-Substrat, Verfahren zu seiner Herstellung und Verfahren zum Detektieren eines Analyten mittels SERS |
EP2319630A1 (de) * | 2009-11-05 | 2011-05-11 | Heidelberger Druckmaschinen AG | Verfahren zum mehrfarbigen, permanenten Lackieren eines Produkts |
US8974869B2 (en) * | 2010-01-26 | 2015-03-10 | Robert Hamilton | Method for improving plating on non-conductive substrates |
CN108212947A (zh) * | 2017-12-27 | 2018-06-29 | 武汉金顿激光科技有限公司 | 激光清洗工艺 |
CN111940423B (zh) * | 2020-08-07 | 2021-07-13 | 武汉金顿激光科技有限公司 | 一种飞机非导电复合涂层的原位激光清洗方法 |
CN115537800A (zh) * | 2022-09-29 | 2022-12-30 | 江苏大学 | 一种金属表面多级超疏水结构的加工系统及其制备方法 |
Family Cites Families (19)
Publication number | Priority date | Publication date | Assignee | Title |
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DE3505750A1 (de) * | 1985-02-20 | 1986-08-21 | Basf Ag, 6700 Ludwigshafen | Campherdithiolen-komplexe und deren verwendung |
DE4318663C1 (de) * | 1993-06-04 | 1994-10-13 | Siemens Solar Gmbh | Verfahren zur Maskierung und Bearbeitung einer Oberfläche eines Substrates |
US5554684A (en) * | 1993-10-12 | 1996-09-10 | Occidental Chemical Corporation | Forming polyimide coating by screen printing |
US5807522A (en) * | 1994-06-17 | 1998-09-15 | The Board Of Trustees Of The Leland Stanford Junior University | Methods for fabricating microarrays of biological samples |
WO1996029629A2 (en) * | 1995-03-01 | 1996-09-26 | President And Fellows Of Harvard College | Microcontact printing on surfaces and derivative articles |
JPH10123315A (ja) * | 1996-10-24 | 1998-05-15 | Canon Inc | カラーフィルターの製造方法 |
CA2268740C (en) * | 1996-11-06 | 2010-07-20 | Sequenom, Inc. | High density immobilization of nucleic acids |
US6048623A (en) * | 1996-12-18 | 2000-04-11 | Kimberly-Clark Worldwide, Inc. | Method of contact printing on gold coated films |
GB9715101D0 (en) * | 1997-07-18 | 1997-09-24 | Environmental Sensors Ltd | The production of microstructures for analysis of fluids |
DE19754978C2 (de) * | 1997-12-11 | 2000-07-13 | Bruker Daltonik Gmbh | Probenträger für die MALDI-Massenspektrometrie nebst Verfahren zur Herstellung der Platten und zum Aufbringen der Proben |
US6159754A (en) * | 1998-05-07 | 2000-12-12 | Intel Corporation | Method of making a circuit edit interconnect structure through the backside of an integrated circuit die |
US6576478B1 (en) * | 1998-07-14 | 2003-06-10 | Zyomyx, Inc. | Microdevices for high-throughput screening of biomolecules |
DE19952323A1 (de) * | 1999-10-29 | 2001-05-03 | Few Chemicals Gmbh Wolfen | Schutzschicht mit antiadhäsiven Eigenschaften |
EP1272768A4 (en) * | 2000-02-23 | 2005-09-07 | Zyomyx Inc | MICROFLUIDIC DEVICES AND ASSOCIATED METHODS |
AU2001268468A1 (en) * | 2000-06-13 | 2001-12-24 | The Trustees Of Boston University | Use of nucleotide analogs in the analysis of oligonucleotide mixtures and in highly multiplexed nucleic acid sequencing |
JP3876684B2 (ja) * | 2000-12-21 | 2007-02-07 | セイコーエプソン株式会社 | カラーフィルタの製造方法、カラーフィルタの製造装置、液晶装置の製造方法、液晶装置の製造装置、el装置の製造方法、el装置の製造装置、材料の吐出方法、ヘッドの制御装置、電子機器 |
US6780233B1 (en) * | 2001-05-24 | 2004-08-24 | Seagate Technology Llc | Wettability improvement of spun-on resist and thermoplastic materials |
EP1425583A1 (en) * | 2001-09-12 | 2004-06-09 | Eidgenössische Technische Hochschule Zürich | Device with chemical surface patterns |
EP1747576A2 (en) * | 2004-05-05 | 2007-01-31 | California Institute of Technology | Capillary lithography of nanofiber arrays |
-
2003
- 2003-03-21 DE DE2003112628 patent/DE10312628A1/de not_active Withdrawn
-
2004
- 2004-03-22 JP JP2006500070A patent/JP2006521538A/ja active Pending
- 2004-03-22 EP EP04722244A patent/EP1606671A2/de not_active Withdrawn
- 2004-03-22 WO PCT/EP2004/002977 patent/WO2004082814A2/de active Application Filing
- 2004-03-22 US US10/550,475 patent/US20060257630A1/en not_active Abandoned
Non-Patent Citations (1)
Title |
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See references of WO2004082814A2 * |
Also Published As
Publication number | Publication date |
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JP2006521538A (ja) | 2006-09-21 |
US20060257630A1 (en) | 2006-11-16 |
DE10312628A1 (de) | 2004-10-07 |
WO2004082814A2 (de) | 2004-09-30 |
WO2004082814A3 (de) | 2005-02-03 |
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