EP1784255A1 - Limite de forme geometrique a surface predeterminee comportant une zone d'ablation, formee sur un materiau poreux monte sur un substrat et ses procedes de fabrication - Google Patents

Limite de forme geometrique a surface predeterminee comportant une zone d'ablation, formee sur un materiau poreux monte sur un substrat et ses procedes de fabrication

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Publication number
EP1784255A1
EP1784255A1 EP05779981A EP05779981A EP1784255A1 EP 1784255 A1 EP1784255 A1 EP 1784255A1 EP 05779981 A EP05779981 A EP 05779981A EP 05779981 A EP05779981 A EP 05779981A EP 1784255 A1 EP1784255 A1 EP 1784255A1
Authority
EP
European Patent Office
Prior art keywords
composite
porous
membrane
hydrophilic
boundary
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Withdrawn
Application number
EP05779981A
Other languages
German (de)
English (en)
Inventor
Keith Wesner
Aaron Spearin
Mark T. Meyering
Douwe Don Haga
Julio Focaracci
Nelson G. Todd
Janusz Wojtowicz
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
3M Innovative Properties Co
Life Technologies Corp
Original Assignee
Applera Corp
3M Innovative Properties Co
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
Application filed by Applera Corp, 3M Innovative Properties Co filed Critical Applera Corp
Publication of EP1784255A1 publication Critical patent/EP1784255A1/fr
Withdrawn legal-status Critical Current

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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
    • 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
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • 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
    • B01J2219/00497—Features relating to the solid phase supports
    • 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
    • B01J2219/00497—Features relating to the solid phase supports
    • B01J2219/00527—Sheets
    • B01J2219/00533—Sheets essentially rectangular
    • 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
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00585—Parallel processes
    • 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
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00596—Solid-phase processes
    • 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
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00639—Making arrays on substantially continuous surfaces the compounds being trapped in or bound to a porous medium
    • B01J2219/00641—Making arrays on substantially continuous surfaces the compounds being trapped in or bound to a porous medium the porous medium being continuous, e.g. porous oxide substrates
    • 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
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00639—Making arrays on substantially continuous surfaces the compounds being trapped in or bound to a porous medium
    • B01J2219/00644—Making arrays on substantially continuous surfaces the compounds being trapped in or bound to a porous medium the porous medium being present in discrete locations, e.g. gel pads
    • 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
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00659—Two-dimensional arrays
    • 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
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00722—Nucleotides
    • 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
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00725—Peptides
    • 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
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00729—Peptide nucleic acids [PNA]
    • 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
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0689—Sealing
    • 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
    • B01L2300/0822—Slides
    • 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
    • B01L2300/16—Surface properties and coatings
    • B01L2300/161—Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
    • B01L2300/165—Specific details about hydrophobic, oleophobic surfaces
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/06—Libraries containing nucleotides or polynucleotides, or derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/04—Libraries containing only organic compounds
    • C40B40/10—Libraries containing peptides or polypeptides, or derivatives thereof

Definitions

  • the present disclosure relates to processes and methods for producing a hydrophobic zone boundary that surrounds a hydrophilic porous material layer mounted on a substrate, the hydrophilic porous material layer containing tortuous channels and pores such that the fluid contained within one hydrophilic layer region does not cross the hydrophobic zone boundary and the articles formed thereby and, more particularly, to processes and methods for producing a hydrophobic zone boundary that separates adjacent regions of a hydrophilic porous material layer mounted on a substrate, the hydrophilic porous material layer containing tortuous channels and pores mounted on a substrate such that a uniform hydrophobic zone boundary layer in the z-direction is formed in the hydrophilic porous material or the removal of the hydrophilic porous material layer from the substrate to form a hydrophilic porous material zone on the substrate, the so formed hydrophilic porous material zone having a predetermined geometric shape and, most particularly, to processes and methods for producing a hydrophobic zone boundary that separates adjacent regions of a hydrophilic porous material mounted on a substrate, the hydrophilic
  • nylon membrane is a hydrophilic porous material, containing tortuous channels and pores for fluid flow and filtration.
  • a membrane surface is less uniform in the z-direction and does not provide as suitable a surface for sealing as a flat film (such as, for example, polyester film/Mylar ® ).
  • the pore structure and hydrophilic character of nylon membrane promotes seepage of liquids in a lateral flow mode, which causes liquid to flow under a gasket. Therefore, a compressed gasket on a hydrophilic nylon membrane surface does not provide a sufficient boundary layer to contain fluid within a gasket sealed area. Because of the porous nylon membrane surface and porous path remaining under the compressed gasket, fluid dispensed within the gasket area, will leak beyond the predetermined boundary layer area.
  • Embossing or etching substrates such as chips, or wafers, with predetermined geometric channels is known in several defined processes.
  • Microporous membrane is placed on the preformed substrates, and then thermally bonded.
  • the surface of the channels is then oxidized to make them hydrophobic. This allows for channels to be predetermined on the substrate, with hydrophobic and hydrophilic regions but none involves bonding the microporous membrane to the support substrate, and then ablating the surface to form separate, hydrophobic zones having a predetermined shape formed thereon to provide the gasket and containment area for the application fluid.
  • 2003018071 1/US-Al, filed- February, 21, 2003 discloses a three dimensional microfluidic device that is formed by placing a membrane between two micropattemed chips.
  • the membrane is positioned to cover the area where channels intersect.
  • the membrane is porous.
  • the chips are formed of plastic, and are thermally bonded under pressure. Reservoirs are formed on the chips at each end of each channel.
  • the channels are created in the chip by use of an embossing master, such as a patterned silicon wafer.
  • the reservoirs are formed by drilling.
  • a hydraulic press is used to emboss both chips, and is also used to thermally bond the chips and membrane under pressure.
  • the surfaces of the channels are oxidized, changing the surfaces from hydrophobic to hydrophilic.
  • 0697377/EP-B1 filed August 18, 1994, discloses a process for production of a glass substrate coated with a patterned Nesa glass membrane which comprises, in sequence: the first step of coating a photoresist on a glass substrate to form a photoresist membrane, exposing the membrane to electromagnetic waves through a mask and then developing the photoresist to form a patterned photoresist membrane on the glass substrate; the second step of forming a Nesa glass membrane on the entire surface of the glass substrate thus provided with the patterned photoresist membrane; and the third step of removing the patterned photoresist membrane together with the Nesa glass membrane thereon from the glass substrate to leave a patterned Nesa glass membrane on the glass substrate.
  • Nesa glass has an electrically conductive surface in the treated area, used for glass electrode measurements. It is not designed for fluid retention on its surface or a hydrophobic boundary, nor affecting a seal.
  • an article and methods of making an article having a hydrophobic zone boundary that surrounds a hydrophilic porous material region or zone the hydrophobic zone boundary being formed on the surface of the hydrophilic porous material, and or a hydrophobic zone boundary that separates adjacent regions of a hydrophilic porous material mounted on a substrate, the hydrophilic porous material containing tortuous channels and pores such that the fluid contained within one hydrophilic region does not cross the hydrophobic zone boundary into any adjacent region and the articles formed thereby.
  • hydrophilic porous material having a hydrophobic zone boundary that surrounds and/or separates adjacent hydrophilic regions formed on the hydrophilic porous material mounted on a composite microarray slide, the hydrophobic zone boundary having a predetermined surface geometric shape for providing a uniform surface for gasket sealing, and fluid retention within the predetermined hydrophilic zone useful for Micro-Analytical Diagnostic Applications.
  • Such composite microarray slides should substantially reduce, if not eliminate, leakage of solutions containing biological polymer (i.e., analytes including but not limited to nucleic acids or proteins), or leakage of reagents that effect the detection of analytes positioned on the surface of the composite microarray slide.
  • the specifically disclosed representative improved composite microarray slides for microarray analysis of the present disclosure include a predetermined surface geometric shape for providing a uniform surface for gasket sealing, and fluid retention within the predetermined geometric area, the predetermined surface boundary geometric shape being, presently preferably, formed by the ablation of the porous polymer membrane attached to the solid substrate for providing a uniform surface for gasket sealing, and fluid retention within the predetermined geometric area.
  • the presently preferred end product is a composite of microporous membrane, presently preferably, nylon microporous membrane operatively mounted on a non-porous substrate, presently preferably, a glass slide by a presently preferably proprietary attachment method, which is disclosed in commonly owned US Patent Application Serial No. 10/410,709 of Keith Solomon et al., filed on July 3, 2001, entitled “Improved Composite Microarray Slides,” or a composite microarray slide.
  • microporous membrane covers one whole slide of the substrate, there are predetermined areas on the surface of the microporous membrane which are active and must be exposed to a variety of chemistries.
  • the microporous membrane is hydrophilic.
  • certain areas of the surface of the composite microarray slides must remain dry.
  • the new and innovative process will selectively "ablate" the pore structure, rendering it non-porous and/or hydrophobic or removing material containing the pore structure entirely from the glass.
  • the presently preferred process comprises representative methods for obtaining hydrophobic/ablated patterns in the composite microarray slide's membrane/composite structure.
  • hydrophobic/ablated patterns define geometric shapes which will effectively isolate any fluid contained within the predetermined geometric boundary.
  • the new and innovative process for producing new and innovative products comprises keeping the hydrophilic area hydrophilic, and interrupting the pore structure around the hydrophilic area for containing a fluid therein. Through the use of interrupted pore structure to form hydrophobic/ablated patterns, the surrounded hydrophilic area can be made into patterns/shapes which are useful for such fluid containment.
  • One object of the present disclosure is to provide commercially useful composite microarray slides having a solid substrate and a porous membrane, the exposed porous membrane surface having a predetermined geometric area defined by hydrophobic boundaries operatively formed thereon which will retain or transport fluids within the predetermined hydrophilic geometric area used in specific representative applications such that the combination produced thereby is useful in microarray applications.
  • Another object of the present disclosure is to provide commercially useful composite microarray slides having a solid substrate and a porous membrane, the exposed hydrophilic porous membrane surface having a predetermined geometric area defined by hydrophobic boundaries operatively formed thereon, the hydrophobic boundaries being operative to transport fluids between various predetermined geometric areas used in specific representative applications.
  • the porous membrane is nylon and the substrate is glass, and the predetermined hydrophilic geometric area is intended to retain liquid hybridization buffers, wash buffers, etc as needed for nucleic acid expression analysis (i.e. microarray).
  • a porous polymer is attached to a solid substrate, and the predetermined hydrophobic boundaries operatively formed thereon are designed to facilitate fluid transport in channels, such as micro channel reactors.
  • the predetermined hydrophobic boundaries operatively formed thereon are patterned for channel chromatography, or membrane based micro fluidics. Many unique products can be envisioned for predetermined geometries formed by membrane ablation on a solid substrate.
  • the immediate objective of nylon ablation with a predetermined geometric shape for a membrane laminated glass substrate is to provide a uniform boundary for gasket placement on the hydrophilic membrane surface.
  • a uniform boundary area predetermined and providing a constant thickness in the z-direction and/or a constant boundary layer caused by either selectively rendering the nylon non-porous or by selective removal of part of the nylon surface from the glass slide is the resultant of the present disclosure.
  • one specific representative aspect of the present disclosure includes a composite device which may be useful for carrying a microarray of biological polymers, the device comprising: a microporous membrane operatively connected to a non-porous substrate having at least one predetermined shaped hydrophilic microporous membrane region, the device having a hydrophobic zone boundary surrounding the at least one predetermined shaped hydrophilic microporous membrane region, the hydrophilic porous material containing tortuous channels and pores.
  • the hydrophobic zone boundary is shaped so that the hydrophobic zone boundary separates adjacent regions of the hydrophilic microporous membrane mounted on the substrate, the hydrophilic microporous membrane containing tortuous channels and pores such that the fluid contained within one hydrophilic region does not cross the hydrophobic zone boundary into any adjacent region.
  • One possible specific application for such innovative is a combination composite microarray slide useful in microarray applications.
  • Another aspect of the present disclosure includes a method of fabricating a composite device comprising the acts of: providing a non-porous substrate; providing a hydrophilic porous membrane; operatively connecting the non-porous substrate to the microporous membrane; and operatively forming at least one predetermined shaped hydrophilic porous material region having a hydrophobic zone boundary.
  • the methods of the present disclosure may be employed to operatively form multiple hydrophobic zone boundaries that separates adjacent regions of a hydrophilic porous membrane on the non-porous substrate, the hydrophilic porous membrane containing tortuous channels and pores such that the fluid contained within one hydrophilic region does not cross the hydrophobic zone boundary into any adjacent hydrophilic region.
  • Figure 1 is a representative depiction of a representative Nylon composite slide with an ablated surface formed from hot die stamping, useful with the present disclosure
  • Figures 2A-2C are representative depictions of the Hot Die Stamping Stages for composite slide stamping, illustrating how to precisely locate and immobilize the composite slide against predetermined reference points (pins) prior to applying the hot die stamp to the composite slide
  • Figures 3A and 3B are a representative graphic depiction of Die heating and containment fixtures for hot die stamping that may be used to form the at least one predetermined shaped hydrophilic porous material region having the hydrophobic zone boundary that separates adjacent regions of a hydrophilic porous material mounted on the substrate of Figure 1
  • Figure 4A is a representative graphic depiction of a prototype hot die stamping dimensions with offset, useful with the present disclosure
  • Figure 4B is a representative graphic depiction of a prototype hot die stamping dimensions without offset, useful with the present disclosure
  • Figure 5 is a representative graphic depiction of the dimension measurements for an ablated nylon
  • analyte or “analyte molecule” refers to a molecule, typically a biological macromolecule, such as a polynucleotide (including, but not limited to, DNA, RNA, cDNA, mRNA, PNA, LNA) or polypeptide, or peptide whose presence, amount, and/or identity is to be determined.
  • a biological polymer may be used as an alternate term for a biological macromolecule.
  • the analyte is one member of a ligand/anti-ligand pair. Alternatively, an analyte may be one member of a complimentary hybridization event.
  • An “Analyte-specif ⁇ c assay reagent” refers to a molecule effective to bind specifically to an analyte molecule.
  • the reagent is the opposite member of a ligand/anti-ligand binding pair.
  • An “array of regions on a solid support” is a linear or two- dimensional a ⁇ ay of preferably discrete regions, each having a finite area, formed on the surface of a solid support.
  • a “microarray” is an array of regions having a density of discrete regions of at least about 100/cm 2 , and preferably at least about 1000/cm 2 .
  • phase inversion process is meant to encompass the known art of porous membrane production techniques that involve phase inversion in its various forms, to produce “phase inversion membranes.”
  • phase inversion membranes it is meant a porous membrane that is formed by the gelation or precipitation of a polymer membrane structure from a “phase inversion dope.”
  • phase inversion dope consists of a continuous phase of dissolved polymer in a good solvent, co-existing with a discrete phase of one or more non-solvent(s) dispersed within the continuous phase.
  • the formation of the polymer membrane structure generally includes the steps of casting and quenching a thin layer of the dope under controlled conditions to effect precipitation of the polymer and transition of discrete (non-solvent phase) into a continuous interconnected pore structure.
  • this transition from discrete phase of non-solvent (sometimes referred to as a "pore former") into a continuum of interconnected pores is generally known as "phase inversion.”
  • phase inversion Such membranes are well known in the art.
  • such membranes and processes will be called “ternary phase inversion” membranes and processes, with specific reference to the ability to describe the composition of the dope in terms of the three major components; polymer, solvent, and non-solvent(s).
  • the presence of the three major components comprise the "ternary" system. Variations of this system include: liquid phase inversion, evaporative phase inversion, thermal phase inversion (where dissolution is achieved and sustained at elevated temperature prior to casting and quenching), and others.
  • ablation refers to the physical change of a part or component of a part by vaporization, crushing, collapse, melting, or other means.
  • Nylon membrane is the part that is ablated during the performance of the process disclosed in the present disclosure.
  • the once porous and hydrophilic nylon membrane becomes non-porous and hydrophobic.
  • Ablation as used in the present application, can result in either a non-porous film, or the loss of substantially all the polymer membrane at the point of ablation.
  • composite slides refers to the product where membrane is adhered to a solid (typically glass) substrate with the use of a surface treatment such as a silane anchor covalently bonded to an epoxy linker attachment chemistry. This surface treatment functions as an adhesive.
  • the epoxy adhered membrane is dried and cured to the glass substrate.
  • Current product configuration is about 3 inches x about 2.5 inches.
  • hydrophobic zone boundary refers to an ablated area operatively positioned on the composite slide's membrane surface defining a boundary, the boundary being defined by the ablated area, the ablated area having any one of a plurality of possible geometrical shapes.
  • the hydrophobic zone boundary is shaped so as to provide a footprint for applying a gasket to the membrane surface of the composite slide when the composite slide is utilized in microarray applications.
  • the gasket and/or boundary layer interface is effective to substantially contain or prevent fluid leakage outside the ablated area defining the hydrophobic zone boundary surrounding the predetermined hydrophilic area. It should be noted, that even without the gasket, there is no leakage evident when liquid is puddled within the hydrophilic area of the microarray that is surrounded by the hydrophobic zone boundary.
  • the fluid is contained by the hydrophobicity of the hydrophobic zone boundary and by the fluids own surface tension.
  • stamping refers to a method of ablating nylon membrane or other porous material to provide a uniform hydrophobic zone boundary.
  • a stamp die with a predetermined dimension is heated to temperatures near or exceeding the melt point temperature of nylon or other porous material.
  • the heated stamp die is placed in contact with the membrane mounted on the substrate, such as, for example, laminated glass. Temperature, pressure, die contact distance, and die contact dwell time, ablates the predetermined surface of the nylon membrane in accordance with the die dimension.
  • stamp dies refers to stamp dies that comprise specific geometric shapes and dimensions. Stamp dies are made of materials that possess high thermal conductivity. Materials include steel, brass, copper, and aluminum and other material having similar thermal properties.
  • Stamp dies can also be comprised of multi materials, or coated with die releasing materials such as chrome plate, dicronite or Teflon ® .
  • Stamp dies have a predetermined geometric shape that is used to provide the hydrophobic zone boundary dimension. Typically, the predetermined die geometric shape that comes into contact with the membrane surface of the composite glass substrate will provide a hydrophobic zone boundary with the same predetermined geometric shape.
  • the term "knife edge dies" refers to dies composed of specific geometric shape and dimensions. A step or recessed area is built into the die surface to provide point or line ablation on the membrane surface of the laminated glass, utilizing conductive and/or radiative heat transfer to the membrane surface of the composite substrate. Knife edge dies are also made of materials that possess high thermal conductivity.
  • laser refers to a highly focused beam of synchronized single-wavelength radiation used to ablate porous material such as, for example, membrane. Table top, commercially available, air cooled, CO 2 lasers were used for ablation of the representative nylon membrane surface on the representative composite glass slides, as described in the present disclosure.
  • vector cutting refers to a type of laser etching. To produce laser etching on a surface, the laser is on continuously at a specified power and frequency, providing the line or point ablation of the membrane coated glass slide. Laser power, speed and frequency will dictate the degree of vector line thickness and depth of surface ablation.
  • rastering cutting refers to another type of laser etching.
  • the laser pulses at a specified dots per inch (dpi), power and speed, providing the ablation of the membrane coated glass slide.
  • the rastering etching method provides uniform depth ablation over a predetermined area of the representative membrane glass slide.
  • the term "leak test” refers to a test method to determine the amount of fluid loss within the hydrophilic area encased by the hydrophobic zone boundary.
  • An apparatus comprised of a composite test slide, a cover glass slide, and a gasket, and a clamping mechanism to apply an even pressure around the gasket is assembled and weighed. The cover glass slide is removed.
  • a predetermined volume of fluid (typically water) is applied within the hydrophilic area encased or surrounded by the hydrophobic zone boundary, and the cover glass is placed over the gasket and clamped under constant pressure.
  • the sample is weighed and placed in an oven at or about 55°C, at or about 18 hours. After about 18 hours at elevated temperature, the sample is weighed, and the fluid weight loss is determined. The percentage of fluid weight loss is calculated. The amount of fluid that escapes from the hydrophilic area encased or surrounded by the hydrophobic zone boundary, determines the effectiveness of the hydrophobic zone boundary to retain fluid within the hydrophilic area encased or surrounded by the hydrophobic zone boundary.
  • microarray slides which comprise a porous nylon or other polymer membrane bound to a solid backing, typically a glass microscope slide.
  • Microarray slides are used in gene sequencing and expression analysis applications where thousands of hybridization assays are performed on the surface of a single microarray slide. It should be understood that the utilization of composite microarray slides is not intended to represent the only possible use of the present innovation but is intended to be merely representative only and that there are a tremendous number of other useful applications for the present innovation and that all such useful applications are intended to be covered by the claims of the present disclosure.
  • the problem to be solved was the failure of the Nylon membrane, which is a hydrophilic porous material, containing tortuous channels and pores for fluid flow and filtration, to provide a suitable surface for containing the liquids positioned on the membrane during certain operations necessary for microarray applications, such as, for example, sealing a membrane surface to prevent the lateral flow of a fluid outside a desired defined area, the membrane surface being less uniform in the z-direction and does not provide as suitable a surface for sealing as a flat film (example: polyester film Mylar ).
  • the pore structure and hydrophilic character of nylon membrane and other known similar porous material promotes seepage of liquids in a lateral flow mode, which allows liquid to flow under a containment barrier that is normally employed during certain operations for microarray applications or other similar operations, such as, for example gaskets. Therefore, a compressed gasket on a hydrophilic nylon membrane surface does not provide a sufficient boundary to contain fluid within a predetermined area sealed by a compressed gasket. Because of the porous nylon membrane surface and porous path remaining under the compressed gasket, fluid dispensed within the gasket sealed area, will leak beyond the predetermined liquid receiving area.
  • a chemical agent that performs the anchor function is applied to the glass slide, rinsed to remove any excess material or reagent, and cured, via an ambient cure, elevated temperature cure, or any combination thereof as would be understood by one skilled in the art.
  • One suitable chemical that functions as an anchor is 3- aminopropyl triethoxysilane.
  • a solution of a suitable chemical reagent that performs the "linker" function is prepared, as follows.
  • One presently preferred chemical reagent that functions as a linker for utilization with the new and improved system of the present disclosure is a Bisphenol A type epoxy, commercially known as Epon 828.
  • any number of curing agents may be used, but at this point, utilization of a polyamide based curing agent, particularly Epikure 31 15, is presently preferred.
  • the two components are mixed, using any suitable means, as would be understood by those skilled in the art.
  • a suitable epoxy- functional silane may be added to the above described mixture of chemical reagents.
  • epoxy-functional silane is 3- glycidopropyltrimethoxysilane.
  • the nylon microporous membrane is then operatively positioned relative to the treated glass slide, restrained in the x- and-y directions, and then oven-cured, as would be understood by those skilled in the art.
  • chemical agents that comprise a surface treatment for providing an attachment layer between the porous membrane and the substrate that would be known to those skilled in the art including, but not limited to, modifications to the silane (anchor) moieties.
  • silane (anchor) moieties may be used for reactivity with glass, including, but not limited to, amines, epoxies, and many others.
  • spin-coating is only one of a plurality of possible methods of applying the surface treatment to the surface of the substrate. Other possibilities include, but are not limited to, drawdown (knife- style), spraying, coating with a slot-die, or equivalents.
  • drawdown knife- style
  • spraying coating with a slot-die
  • the presently perceived primary advantage of spin-coating is the resulting high uniformity of application of chemical agent comprising the surface treatment on the micro scale.
  • the membrane may be charged or uncharged and the pore size and thickness of the membrane can be manipulated to any desired range, as would be understood by one skilled in the art.
  • the membrane may or may not contain pigment for modification of optical surface reflectance properties.
  • Nylon/Glass Composite slides useful as a composite microarray slides for carrying a microa ⁇ ay of biological polymers was carried out as follows in accordance with the Solomon et al., application.
  • This representative Example described the process for producing a sample batch of the nylon/glass composite slides.
  • the representative nylon/glass composite slides which were produced were comprised of a thin ( ⁇ 2 mil) layer of porous nylon membrane operatively bound to the surface of a glass microscope slide. Such slides have proven operable as composite microarray slides useful for carrying a microarray of biological polymers.
  • the representative process was initiated by dissolving one packet of NoChromix ® (Godax Labs, Inc) into about 2.5L of concentrated sulfuric acid, then stirring thoroughly until all crystals were dissolved to produce a cleaning solution.
  • the previously prepared cleaning solution was poured into a glass dish (Thermo Shandon model 102), and allowed to sit for about 10 minutes. Glass microscope slides were placed into a 20 slide rack and then immersed in the cleaning solution, above, for about 30 minutes, then transfe ⁇ ed to another dish filled with about 18m ⁇ DI water where they remained for about 20 minutes. The slides were then dipped briefly in HPLC grade denatured ethanol (Brand-Nu #HP612) and then silanated by the procedure described below. Alternately, the slides may be cleaned with an about 1 wt% solution of Alconox in DI water; air agitated for about 30 minutes, or a heated ultrasonic bath, followed by about a 30 minute sparge with frequently refreshed baths of 18m ⁇ DI water.
  • the slides were silanated by the following representative procedure: First, an about 100 mL solution of about 95% ethanol and about 5% water (percent by volume) was prepared. Then, about 2 mL of 3- aminopropyldimethylethoxysilane (United Chemical Technologies #A0735) was added to the above solution, mixed thoroughly, and allowed to sit for about 5 minutes. After the preceding about 5 minute activity was complete, the resulting solution was poured into glass dish, and the slides were immersed therein for about 2 minutes. The slides were then removed from the silane solution, dipped into a dish containing ethanol for about 7 seconds, and removed from the dish. The slides were then placed into an oven for about 10 minutes at about 110° C, and allowed to finish reacting overnight.
  • 3- aminopropyldimethylethoxysilane United Chemical Technologies #A0735
  • a representative Bisphenol A "linker” solution was made by adding the following to a 250 mL Erlenmeyer flask and mixing thoroughly after each step in which a new ingredient was added: about 10 grams Epon 828 (a Bisphenol A type epoxy resin); and about 34 grams Xylene. In a separate 250 mL Erlenmeyer flask, the following were also added: about 4.1 grams Epikure 3115 (a polyamide based curing agent); about 34 grams Xylene; and about 1.8 grams 3-glycidopropyltrimethoxysilane.
  • the contents of the first flask were then poured into the second flask, sealed, and agitated with a lab stirrer for about an additional about 15 hrs at about 60° C.
  • the resultant solution from the combination of the two flasks described above resulted in an about 12 wt% Bisphenol A "linker" solution.
  • a single cleaned and silanated slide was then placed on a spin coater (Specialty Coating Systems model P6708). Surface was flooded with the epoxy solution prepared above, then allowed to spin at the following cycle:
  • the wet-as-cast porous nylon membrane used had been cast, quenched, and washed with DI water, but had not yet been exposed to a drying step, hence the term "wet-as-cast.”
  • the wet-as-cast porous nylon membrane had a thickness of approximately 1.5 mils, a nominal pore size less than about 0.2 micron, and a target initial bubble point in water of about 135 PSI (once dried).
  • the base polymer for this wet-as-cast porous nylon membrane is Vydyne 66Z nylon (Solutia, Inc), which is a high molecular weight nylon that is preferentially terminated by amine end groups.
  • wet-as-cast porous nylon membrane was applied to the treated slides. care was taken to ensure removal of any air bubbles between the wet-as-cast porous nylon membrane and each slide.
  • the wet-as-cast porous nylon membrane was flattened onto each slide and all wrinkles were removed. Once positioned on the slides, the wet-as-cast porous nylon membrane was clipped into position, as is known in the art. The entire assembly was then heated in a convection oven at about 1 10° C for about 45 minutes. After heating, the excess, now dried, porous nylon membrane was removed from the slides by trimming, as is known in the art. Following trimming, the slides were allowed to sit overnight, in order for the epoxy resin to further cure.
  • a solution of 4x SSC sodium salt, sodium citrate
  • 4x SSC sodium salt, sodium citrate
  • the slides were placed into a Tupperware container, SSC solution was poured on top of the slides, and the container was sealed. The container was then placed in a hybridization oven at about 60° C for a minimum of about 12 hours with gentle rocking. Upon removal from the solution, all the membrane components of the composite slides were found to be securely bonded to the substrate component, with no delamination of the membrane from the substrate. The slides that were exposed for a longer period at 60° C, in excess of 72 hours, also showed no delamination of the nylon from the substrate.
  • DMF n-dimethylformamide
  • the slides were allowed to sit at room temperature for a minimum of about 6 hours, then removed and rubbed firmly. After the above treatment, the slides exhibited no loss of adhesive strength of the bond between the membrane and the substrate after immersion in
  • the improved composite microarray slides useful for carrying a microarray of biological polymers on the surface thereof and, more particularly, to an improved composite microarray slide having a porous membrane formed by a phase inversion process effectively attached by covalent bonding or hydrogen bonding through chemical agents that comprise a surface treatment to a substrate the surface treatment preparing the substrate to sufficiently bond to the microporous membrane through the attachment layer formed therebetween resulting from the surface treatment such that the combination produced thereby is useful in microarray applications was completed, the need to develop methods for effectively containing microarray fluid chemistry within a predetermined hydrophilic region or zone on the membrane surface of the improved composite microarray slides, such as, for example, a nylon coated, composite microarray slide was soon recognized.
  • nylon membrane is a hydrophilic porous material, containing tortuous channels and pores for fluid flow and filtration.
  • the hot die stamping process is accomplished by placing a heated die having a predetermined geometric shape unto the porous polymeric surface such as, for example, nylon surface of the composite microarray slide for a specified dwell time. Once positioned, the die is heated to a temperature at or near the melt point temperature of the nylon polymer membrane. The nylon polymer membrane surface then vaporizes or melts, leaving a hydrophobic boundary zone having a predetermined geometric shape that surrounds at least one hydrophilic zone on the surface of the composite slide, the predetermined geometric shape of the hydrophilic zone being defined by the shape and dimensions of the die used in the stamping process.
  • the hydrophobic boundary zone geometric shape is consistent with the die geometric shape and surface area that comes in contact with the nylon polymer membrane on the surface of the composite slide.
  • FIG. 1 illustrates a representative composite microarray slide produced using a representative hot die stamping process comprises positioning a heated die in a fixed position in the x and y axis above the porous composite substrate which is precisely located and immobilized or restrained in a suitable fixture (see Figures 2A-2C).
  • the representative composite microarray slide is restrained by the fixture positioned below the die (see Figure 2).
  • the representative composite microa ⁇ ay slide is restrained in the x and the y plane, and is referenced in the same position at the start of each stamping.
  • the representative composite microarray slide is restrained in order to maintain dimension boundaries for die placement on the representative composite microa ⁇ ay slide.
  • the representative composite microa ⁇ ay slide is restrained by conventional means, such as, for example, vacuum, tension springs, and or reference pins etc.
  • a hot die upon restraining the representative composite microa ⁇ ay slide, a hot die will traverse along the z axis until the die comes in contact with the upper surface of the representative composite microa ⁇ ay slide.
  • a positive stop may be used to prevent the die from crushing the porous composite substrate, and maintain a predetermined placement in the z- direction.
  • the dies that are utilized in the representative process have a predetermined shape for su ⁇ ounding a predetermined surface area of the composite microarray slide to be isolated.
  • the dies and die fixture are heated to a predetermined temperature at or near the melting point of the porous material attached to the non porous substrate. Temperature control of the die can be maintained within about 1° Fahrenheit.
  • Figures 2A, 2B and 2C are diagrams of the production stages for providing a consistent positioning of the representative composite microa ⁇ ay slide that are used in the hot die stamping process.
  • the various production stages are used to position the representative composite microa ⁇ ay slide along the x and y axis.
  • reference pins position the representative composite microa ⁇ ay slide on the x-axis.
  • a clip spring applies pressure along the axis to maintain constant pressure during hot die stamping.
  • Vacuum holes are operatively position thereon for cooperating with a vacuum suction cup, as illustrated in Figure 2B.
  • a vacuum is applied to the center of the bottom of the representative composite microa ⁇ ay slide to maintain position of the representative composite microa ⁇ ay slide during hot die stamping.
  • the reference pins are used to keep the representative composite microa ⁇ ay slide stationary.
  • the clip spring is positioned in the corner to keep constant pressure along the x and y axis during stamping, thus maintaining position of the glass slide during the ablation process.
  • insulation is added during this stage. The insulation is consistent to the hot die stamping.
  • the heat insulation includes, but is not limited to, ceramic, layered composite, such as mica, or high heat resistance material.
  • This heat insulation keeps the temperature in the insulated area, focused on the representative composite microa ⁇ ay slide ablation area
  • a hot die will traverse along the z axis until the die comes in contact with the upper surface, the surface having the porous material, of the representative composite microa ⁇ ay slide.
  • a positive stop will prevent the die from crushing the representative composite microa ⁇ ay slide, and maintain a predetermined placement in the z-direction.
  • the dies utilized in this operation have a predetermined shape and size.
  • the dies and die fixture are heated to a predetermined desired temperature at or near the melting point of the porous material on the upper surface of the representative composite microa ⁇ ay slide.
  • the die temperature control was maintained within about 1° Fahrenheit.
  • Figure 3 shows die staging and heating of the representative dies used to effectuate the ablation of the upper surface of the representative composite microa ⁇ ay slide.
  • the dies are secured in a dovetail steel plate or similar fixture. The fixture is heated by adding electrical heating cartridges. Because the die is in direct contact with the heating block, temperature uniformity within the die is approximately consistent.
  • the illustrated dies are made according to predetermined shapes in order to form the hydrophobic zone boundary that separates adjacent regions of a hydrophilic porous material mounted on representative composite microa ⁇ ay slides.
  • the dies are presently preferably made of highly conductive materials, including, but not limited to, brass, copper, steel, aluminum and chrome etc.
  • any material that can maintain temperatures or transfer heat, at or near the melting point of the porous material that comprises the upper surface of the representative composite microa ⁇ ay slide can be utilized.
  • Typical hot dies used in ablation of porous substrates are illustrated in Figures 4 and 4B. Dies can have flat surface of have a contact surface that provides an offset. Offset dies will provide degrees of ablation on the composite surface. The knife edge (offset die), will make contact with the upper surface of the representative composite microa ⁇ ay. This allows the knife edge to provide complete ablation along the knife edge axis.
  • representative stamping dies are illustrated having representative detailed dimensions and sizes for a representative die used for porous material ablation.
  • the hot die stamping ablates the surface of the porous nylon membrane, leaving a predetermined geometrically shaped impression, upon retraction of the stamp die.
  • the die dimensions are co ⁇ elated to a specifically desired finished ablated porous material shaped surface designed to su ⁇ ound the desired predetermined hydrophilic porous material mounted on a substrate, the impression defining a hydrophobic zone boundary.
  • the hydrophobic zone boundary dimension measurements can be measured using an optical comparator, or computer optical scanner.
  • Figure 5 illustrates a representative composite microa ⁇ ay slide with an ablated surface that was formed using hot die stamping. The white area represents where the hot die came in contact with the porous material that forms the upper surface of the representative composite microa ⁇ ay.
  • the dimension of the hot die can be changed, based on die contact surface dimension, contact time and temperature of the die surface and other appropriate factors in order to define the hydrophobic zone boundary that su ⁇ ounds the shaped surface designed to su ⁇ ound the desired predetermined hydrophilic porous material mounted on a substrate.
  • the white surface represents the ablated hydrophobic zone boundary area of the porous surface on the total representative composite microa ⁇ ay slide area.
  • the grey area represents the desired predetermined hydrophilic microporous surface not ablated on the total upper surface of the representative composite microa ⁇ ay slide.
  • the white surface area shows the dimension measurements for the ablated porous material surface wherein the grey surface represents the unablated area of the upper surface of the representative composite microa ⁇ ay slide.
  • the various dimensions of the illustrated composite microarray slide can be manipulated such that the various measurements can determine the hydrophobic zone boundary placement, inside hydrophobic zone boundary dimensions, and thickness of the ablated area.
  • the white surface area indicates the ablated hydrophobic zone boundary area of the porous surface on the total representative composite microa ⁇ ay slide surface area.
  • a knife edge heat stamp product is illustrated in Figure 8.
  • the grey area indicates the porous material surface not ablated on the total composite microa ⁇ ay slide area
  • the white portion represents the ablated area wherein at least a portion of the remaining porous material remains positioned on the non porous substrate and the black lines in and around the surface of the ablated hydrophobic zone boundary representative areas that are completely ablated/removed from the surface of the porous material and form line channels on the representative composite microa ⁇ ay slide.
  • Example 1 Control Slide: A control sample was conducted along with the test slide samples. The control sample consisted of two glass slides containing the gasket and test fluid only. The control sample was tested and compared with the representative porous composite microa ⁇ ay slide having ablated hydrophobic area boundary.
  • control sample determined if the gasket and test apparatus is able to contain the fluid.
  • the control samples established a functional baseline; i.e. fluid leakage for the gasket only.
  • Figures 6 and 7 illustrate the test apparatus used to conduct the leak tests.
  • a control slide is a plain glass slide with no membrane attached. The intention of the control slide is to function as control in the leak test. The control slide is not porous and has no hydrophilic zone, therefore, it should provide a baseline for the leak test. Table 1. Control slide percent fluid loss for a leak test
  • Example 2 Composite slide with no ablation This is the composite slide described in Solomon, et al. which has a microporous membrane attached to a glass substrate, but has no ablated areas. The intention of the composite slide is to demonstrate the problem of leakage in a microa ⁇ ay application where a gasket is applied as a sole means of fluid retention in the hydrophilic zone. Upon repeated leak testing, it was discovered that the non ablated substrate membrane was bone dry, after removal of the leak test assembly. It is therefore concluded that all the water (100%) was evaporated and lost from the test apparatus Table 2: Non ablated composite slide leak test data
  • the dies are heated at or near the melt point temperature of the polymer surface for effective ablation and creation of the predetermined hydrophobic zones.
  • the typical operating temperatures for dies used to stamp nylon covered representative composite microa ⁇ ay slides are from about 600 to about 850° Fahrenheit.
  • the dies will expand as die surface temperature increases. This thermal expansion is dependent on the particular type of die material. As would be expected, the die expansion is in the x and y axis and is typically uniform across the surface of the die.
  • Hot die ablation of a representative porous composite microa ⁇ ay slide can be made in any one of a plurality of dimensions; thus defining a hydrophobic boundary around a hydrophilic composite porous membrane zone.
  • the ablated area is defined by the die dimension, and placement on the representative porous composite microa ⁇ ay slide surface. Placement of the ablated area on the representative porous composite microa ⁇ ay slide surface is defined by the die process staging and the representative porous composite microa ⁇ ay slide surface area.
  • One simple and effective method for determining the ablated hydrophobic boundary capability for limiting fluid loss outside the hydrophobic boundary zone comprises applying a fluid within the hydrophilic zone su ⁇ ounded by the ablated hydrophobic boundary area, which will provide fluid retention up to the point where the mass of water exceeds the microporous membrane capacity to contain the fluid, would be understood by those skilled in the art.
  • Another method for determining the ablated hydrophobic boundary capability for limiting fluid loss from the su ⁇ ounded hydrophilic zone outside the hydrophobic zone includes performing a gasket leak test. The gasket leak test is initiated by placing a predetermined amount of fluid, typically water, within the predetermined hydrophilic porous material zone su ⁇ ounded by hydrophobic boundary as zone defined by the area where the porous material was ablated.
  • a gasket is placed on the surface of the ablated zone of the representative porous composite microarray slide and then sealed with a glass substrate on the top side, under constant compression.
  • the gasketted representative porous composite microa ⁇ ay slide having the ablated porous material boundary surrounding the containment fluid is heated to about 55°C for a predetermined time increment.
  • the weight of the gasket seal test apparatus is measured prior to fluid being added to the containment area, then with fluid containment prior to heating, and finally with whatever contained fluid remains after heating. The weight differences between the gasket seal test apparatus at these times determines the amount of fluid that escapes/evaporates during the test.
  • Gasket are generally difficult to manufacture especially flat gaskets and can have substantial variation in both the cutting of the gasket to achieve a particular size and also in the placement of the gasket on the surface of a representative composite microa ⁇ ay slide.
  • the combination of gasket placement e ⁇ or and gasket manufacturing e ⁇ or can be typically as high as about +/- 0.020 in.
  • Hot die stamping with a flat surface die Hot die stamping of a nylon composite slide is achieved by using a rectangle steel die as described in figure 4b heated at or around 790°F and having a contact time of around 5 seconds on the composite nylon slide surface (same composite slide construction as example 2; with the exception that the hot die creates an ablated hydrophobic rectangle with defined geometry).
  • the dies are heated at or near the melt point temperature of the porous polymer surface for effective ablation, loss of pore structure, and creation of the predetermined hydrophobic zones.
  • the typical operating temperatures for dies used to stamp nylon micro-a ⁇ ay slides are from about 600 to about 850° Fahrenheit. During the process, the dies will expand as die surface temperature increases.
  • Hot die ablation of a porous composite substrate can be made in any one of a plurality of dimensions; thus defining a hydrophobic boundary around a hydrophilic composite porous membrane zone.
  • the ablated area is defined by the die dimension, and placement on the composite porous substrate surface. Placement of the ablated area on the composite surface is defined by the die process staging and composite surface area.
  • a method for determining the ablated membrane area capability for limiting fluid loss from the su ⁇ ounded hydrophilic zone outside the hydrophobic zone includes performing a gasket leak test.
  • the gasket leak test has been described previously.
  • Figures 6 and 7 illustrate the test apparatus used for the leak test.
  • Leak testing results are considered acceptable if the tested ablated slide percent leakage is less than about 10%, and control slides do not exhibit failure.
  • the about 10% fluid loss is based on acceptance of microa ⁇ ay test fluid loss limits. Measurements were made to determine the precision of both the placement and the internal dimensions of the hydrophobic ablated zone. This was done to ensure that the desired defined geometry was successfully produced on the composite slide. The measurements are taken by using an optical comparator or a camera optical measurement device. To verify the hydrophobic zone placement on the composite slide, a series of measurements was conducted from the reference edges of the composite slide (refer to Figure 1). Measurements were made from the x-axis and y-axis to the respective parallel boundaries of the hydrophobic ablated zone.
  • Two measurement locations were chosen for the x-axis placement and two for the y-axis placement.
  • a total of 17 slides were measured in each of the four reference locations.
  • a mean and standard deviation were calculated for each of the four reference locations. The worse case standard deviation was chosen to represent the maximum offset variation relative to the reference edges.
  • To verify the hydrophilic zone dimensional area on the composite slide a series of measurements was conducted from the inner edges of the hydrophobic zone (fluid containment area, refer to Figure 1). Measurements were made of the length and the width of the fluid containment area. Two measurement locations were chosen for the length and two for the width. A total of 17 slides were measured in each of the four reference locations. A mean and standard deviation were calculated for each of the four reference locations.
  • test results for composite microa ⁇ ay slides not having their upper surfaces altered in accordance with the innovations of the present disclosure indicated and almost total loss of fluid, as was also expected.
  • test results for the composite microa ⁇ ay slides having their surfaces altered in accordance with the above example 3 allowed only a two- three percent loss of fluid. This is believed to be significant in that is somewhat less than the 10 percent loss considered acceptable.
  • Knife Edge Dies for conducting surface ablation and hydrophobic zone boundary definition Knife edge dies can be used to define the ablated hydrophobic zone boundary on the representative porous composite microarray slide. Knife edge dies have a recessed area on the contact surface of the die. This recess allows for different degrees of ablation of the nylon surface of the representative porous composite microa ⁇ ay slide. Total porous material surface ablation is accomplished by the die areas that first directly contact the nylon surface, while the recessed die area, in close proximity to the nylon surface, accomplishes partial ablation of the nylon surface (refer to Figure 4A). The areas between the knife edges of the recessed die provide thermal energy to at least partially ablate the porous nylon surface, i.e.
  • the inside surface of the recessed die provides a very uniform ablation, thus providing a substantially uniform hydrophobic zone boundary for gasket placement.
  • Areas of the die (non recessed areas) that first come in direct contact with the porous nylon surface comprise relatively thin lines, or points, which typically ablate the total surface of the porous nylon surface that they contact, thus creating channels or grooves in the non porous substrate underlying the nylon porous membrane surface.
  • Recessed dies used as described above can be made of aluminum, brass, copper, or other highly thermal conductive material. The process for recessed ablation is substantially the same as described for hot die stamping.
  • a brass die was chosen, along with a copper stage. The brass die was fabricated with .003" recess. The copper stage was fabricated with recessed insulation built into the stage (refer to figure 4A).
  • Table 4 Leak testing for a recessed Brass die with a recessed insulated Copper stage
  • Single-wavelength radiation Laser light can also be used to completely ablate or partially ablate the porous material surface on the composite slide.
  • Table top, commercially available, air cooled, 35 watt CO 2 lasers can be used for the ablation of the nylon membrane surface on the representative porous composite microarray slide.
  • the laser can replicate the effect of hot die stamping with a rastering laser cutting, or ablate the entire nylon surface on the representative porous composite microa ⁇ ay slide with vector cutting.
  • Vector cutting is a type of laser etching as specified by the commercially available laser unit.
  • Vector laser etching is defined as the laser synchronized light source emitting continuously on at a specified power and frequency, providing the line or point substantially complete ablation of the nylon membrane covered representative porous composite microa ⁇ ay slide.
  • Laser power, speed and frequency will dictate the vector line thickness dimension and the depth of ablation of the nylon porous material surface on the composite slide.
  • Another type of laser etching is rastering cutting.
  • DPI, power and speed provide the energy to ablate the porous nylon membrane surface of the representative porous composite microa ⁇ ay slide.
  • the rastering etching methods provides uniform depth ablation over the predetermined area of the representative porous composite microarray slide, similar to that achieved by the previously described hot die stamping.
  • Computer graphing software has been used to determine placement of the vector or rastering cutting on the porous material surface on the representative porous composite microarray slide, and is the laser instrument method for defining placement of the ablated boundary zone in the x and y direction.
  • Laser vector ablation allows lines to be cut into the porous material surface on the representative porous composite microarray slide as well as to and into the nonporous substrate.
  • the vector line can be cut to the surface of the support substrate, thus completely ablating the nylon at the point of contact of the laser beam.
  • the lines in the porous material and the support substrate act as barrier walls or channels to retain fluid within the predetermined hydrophilic zone su ⁇ ounded by the hydrophobic zone boundary of the representative porous composite microa ⁇ ay slide.
  • a gasket is placed over the vector cut ablated lines for testing in the leak test.
  • Vector cut ablation lines formed by laser vector cutting can range from one two to as many as seven or as many as may be required for a specific application within a defined hydrophobic zone boundary to provide the necessary boundary for fluid containment.
  • a gasket is placed over the vector cut ablated lines for sealing the circumference of the hydrophilic zone su ⁇ ounded by the hydrophobic zone boundary.
  • Figure 9 is a schematic illustrating ablated vector lines placed on a representative porous composite microa ⁇ ay slide using such lasers.
  • Example 5A, 5B and 5C Surface ablation using laser vector line cutting Laser cutting samples generated by the Epilog laser were evaluated for dimensional tolerances. Vector cutting was conducted under the following Epilog ® laser process settings: Table 5: Vector etching laser process settings
  • Example 6 Surface ablation using laser rastering etching Laser cutting samples generated by the Epilog laser were evaluated for dimensional tolerances. Rastering cutting was conducted under the following Epilog ® laser process settings shown in Table 7 below: Table 7: Rastering etching laser process settings
  • Example 7 Surface ablation using laser rastering etching and vector cutting
  • Laser cutting samples generated by the Epilog laser were evaluated for dimensional tolerances. Ablation of the microa ⁇ ay surface was done with rastering etching and vector cutting. The hydrophobic ablated zone was first etched with a laser using rastering etching. Vector cutting with the laser defined Jhe hydrophic ablation zone. In some samples, multiple vector lines were placed within the ablated hydrophic zone. Raster cutting alone to ablate and form the hydrophobic surface on the micro-a ⁇ ay was shown to have a high degree of variability when using the leak test for gasket functionality. Variability of the membrane thickness, glass and adhesive coating, in addition to the laser process variability will result in an inconsistent cut width and depth on the hydrophobic ablated area.
  • the hydrophobic ablated area is flatter than the non-ablated surface of the micro-a ⁇ ay slide, however, variation in cut width and depth is observed.
  • the raster cut will provide the flat surface required for applying the gasket in the hydrophic area on the micro-a ⁇ ay slide.
  • Vector cutting was added to the raster ablation cutting to improve the hydrophobic ablated area width and placement dimensions.
  • Vector cutting along the inner and outer borders of the hydrophobic area improved ablated area dimension placement.
  • the hydrophobic ablated zone was first etched with laser using rastering etching. Vector cutting with the laser defined the hydrophic ablation zone. In dual raster and vector cut samples, multiple vector cut lines were added within the ablated hydrophic zone. Table 9: Rastering etching laser process settings
  • the leak test results indicated that the innovation of the present disclosure was successful in meeting the 10% fluid leak target.
  • the processes for forming hydrophobic boundaries su ⁇ ounding hydrophilic areas have proven extremely successful.
  • the innovative ablation techniques applied to the composite slides result in well controlled, predetermined geometric shaped boundaries formed on the slides, and have the beneficial capabilities of providing zones for containing fluid, effectively forming ba ⁇ iers to prevent fluid leakage when used in conjunction with a sealing apparatus such as a gasket..
  • the ablated zone(s) further have a hydrophobic characteristic, which beneficially help to direct or contain aqueous liquid to the more hydrophilic porous structure.
  • the ablated zone(s) have well defined geometries, and (in conjunction with proper fixturing devices) can be placed reproduceably and accurately in predetermined locations on a representative composite slide, which results in an improved product useful for microa ⁇ ay applications. While the shapes of the hydrophilic zones illustrated herein have been square or rectangular in shape, it should be understood that the innovations described herein are not limited to any specific shape and that all possible geometric shapes are believed possible in the practice of these innovations. While the articles, apparatus and methods for making the articles contained herein constitute prefe ⁇ ed embodiments of the invention, it is to be understood that the disclosure is not limited to these precise articles, apparatus and methods, and that changes may be made therein without departing from the scope of the disclosure which is defined in the appended claims.

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Abstract

La présente invention concerne des processus et des procédés destinés à la production d'une limite de zone hydrophobe qui entoure une couche de matériau poreux hydrophile montée sur un substrat, la couche de matériau poreux hydrophile contenant des canaux sinueux et des pores de façon que le fluide contenu dans une région de la couche hydrophile ne traverse pas la limite de zone hydrophobe. L'invention concerne également des articles formés selon l'invention. L'invention concerne plus précisément des processus et des procédés destinés à la production d'une limite de zone hydrophobe qui sépare des régions adjacentes d'une couche de matériau poreux hydrophile montée sur le substrat, la couche de matériau poreux hydrophile contenant des canaux sinueux et des pores étant montée sur le substrat de façon qu'une couche limite de zone hydrophobe uniforme dans la direction z soit formée dans le matériau poreux hydrophile ou que le retrait de la couche de matériau poreux hydrophile du substrat forme une zone de matériau poreux hydrophile sur le substrat, la zone de matériau poreux hydrophile ainsi formée possédant une forme géométrique prédéterminée, de façon que la combinaison ainsi obtenue soit utile dans des applications de microréseaux et d'autres application. L'invention concerne également des produits de ces processus et procédés.
EP05779981A 2004-05-13 2005-05-13 Limite de forme geometrique a surface predeterminee comportant une zone d'ablation, formee sur un materiau poreux monte sur un substrat et ses procedes de fabrication Withdrawn EP1784255A1 (fr)

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US57144604P 2004-05-13 2004-05-13
PCT/US2005/016781 WO2005113131A1 (fr) 2004-05-13 2005-05-13 Limite de forme geometrique a surface predeterminee comportant une zone d'ablation, formee sur un materiau poreux monte sur un substrat et ses procedes de fabrication

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EP1784255A1 true EP1784255A1 (fr) 2007-05-16

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EP (1) EP1784255A1 (fr)
AU (1) AU2005245425A1 (fr)
BR (1) BRPI0511003A (fr)
WO (1) WO2005113131A1 (fr)

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GB201020574D0 (en) 2010-12-03 2011-01-19 Univ Glasgow Assay assembly and methods

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AUPN239395A0 (en) 1995-04-12 1995-05-11 Memtec Limited Method of defining an electrode area
AU2460399A (en) 1998-01-20 1999-08-02 Packard Bioscience Company Gel pad arrays and methods and systems for making them
US20030113528A1 (en) 1999-09-17 2003-06-19 Wilson Moya Patterned porous structures
US20030099949A1 (en) 2001-10-05 2003-05-29 Surmodics, Inc. Arrays having clustered arrangements and methods of making and using
US7195872B2 (en) 2001-11-09 2007-03-27 3D Biosurfaces, Inc. High surface area substrates for microarrays and methods to make same
JP2006515065A (ja) 2002-08-16 2006-05-18 ディシジョン バイオマーカーズ インコーポレイテッド 蛍光配列の読み取り
US6913931B2 (en) 2002-10-03 2005-07-05 3M Innovative Properties Company Devices, methods and systems for low volume microarray processing

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See references of WO2005113131A1 *

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WO2005113131A1 (fr) 2005-12-01
AU2005245425A1 (en) 2005-12-01
BRPI0511003A (pt) 2007-11-20

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