CA2213694C - Composite waveguide for solid phase binding assays - Google Patents

Composite waveguide for solid phase binding assays Download PDF

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Publication number
CA2213694C
CA2213694C CA002213694A CA2213694A CA2213694C CA 2213694 C CA2213694 C CA 2213694C CA 002213694 A CA002213694 A CA 002213694A CA 2213694 A CA2213694 A CA 2213694A CA 2213694 C CA2213694 C CA 2213694C
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waveguide
biosensor
optical material
film
refractive index
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CA2213694A1 (en
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W. Monty Reichert
James N. Herron
Hsu-Kun Wang
Douglas A. Christensen
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University of Utah Research Foundation UURF
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366Apparatus specially adapted for solid-phase testing
    • G01N33/54373Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/645Specially adapted constructive features of fluorimeters
    • G01N21/648Specially adapted constructive features of fluorimeters using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/7703Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator using reagent-clad optical fibres or optical waveguides
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03Cuvette constructions
    • G01N21/05Flow-through cuvettes

Abstract

A step-gradient composite waveguide (100) for evanescent sensing in fluorescent binding assays comprises a thick substrate layer (102) having one or more thin film waveguide channels (104) deposited thereon.

Description

COMPOSITE WAVEGUIDE FOR SOLID PHASE BINDING ASSAYS

BACKGROUND OF THE INVENTION
Field of the Invention: The invention relates to apparatus for solid-state biochemical binding assays, and especially to optical structures utilizing evanescent sensing principles for use in such apparatus and assays.
State of the Art: Immunoassays exploiting the properties of an optical technique known as total internal reflection (abbreviated TIR) are proving to be a valuable tool for detection of analytes at concentrations of 10-10 to 10-13 molar or below, without a wash step. RThen a light beam traveling in a waveguide is totally internally reflected at the interface between the waveguide and an adjacent medium having a lower refractive index, a portion of the electromagnetic field of the TIR
light penetrates shallowly into the adjacent medium. This phenomenon is termed an "evanescent penetration" or "evanescent light". The intensity of evanescent light drops off exponentially with distance from the waveguide surface.
Binding assays in general are based on the strong affinity of a selected "capture" molecule to specifically bind a desired analyte. The capture molecule/analyte pair can be an antibody/antigen pair or its converse, a receptor/ligand pair or its converse, etc, as known in the art. In a fluorescent binding assay, the binding of the analyte to the antibody is monitored by a tracer molecule which emits fluorescent light in response to excitation by an input light beam.
One of several possible schemes for exploiting the properties of evanescent light fields for fluorescence measurements is as follows. If an antibody is immobilized on an optical structure in which a light beam is being propagated by TIR, the resulting evanescent light can be used to selectively excite tracer molecules that are bound (whether directly or indirectly) to the immobilized antibody.
Tracer molecules free in solution beyond the evanescent penetration depth are not excited, and therefore do not emit fluorescence. For silica-based optical materials or optical plastics such as polystyrene, with the adjacent medium being an aqueous solution, the evanescent penetration depth is generally about 1000 to 2000 A
(angstroms).
The amount of fluorescence is thus a measure of the amount of tracer bound to the immobilized capture molecules. The amount of bound tracer in turn depends on the amount of analyte present, in a manner detezmined by the specifics of the immunoassay procedure.
U.S. Patents Nos. RE 33,064 to Carter, 5,081,012 to Flanagan et al, 4,880,752 to Keck, 5,166,515 to Attridge, and 5,156,976 to Slovacek and Love, and EP publications Nos. 0 517 516 and 0 519 623, both by Slovacek et al, all disclose apparatus for immunoassays utilizing evanescent sensing principles.
Desirably, an immunosensor should be capable of accurately and repeatably detecting analyte molecules at concentrations of 10-13 M (molar) to 10-15 M
and preferably below. At present, such sensitivity is not believed to be available in a commercially practical and affordable immunosensor. Also desirably, an immunosensor should provide multiple "channels", that is, the capacity for measuring multiple analytes and multiple measurements of the same analyte, on the same waveguide substrate. Such an immunosensor would allow both self-calibration with known standards, and screening for a panel of different analytes selected for a particular differential diagnostic procedure.
One approach to improving the sensitivity (lowering the detection limits) of fluorescent immunosensors, proposed by Ives et al. (Ives, J.T.; Reichert, W.M.;
Lin, J.N.; Hlady, V.; Reinecke, D.; Suci, P.A.; Van Wagenen, R.A.; Newby, K.;
Herron, J.; Dryden, P. and Andrade, J.D. "Total Internal Reflection Fluorescence Surface Sensors" in A.N. Chester, S. Martellucci and A.M. Verga Scheggi Eds.
Ontical Fiber Sensors, NATO ASI Series E, Vol 132, 391-397, 1987), is to use waveguides which are very thin, perhaps about l m in thickness. Such thin waveguides may provide higher evanescent intensity and a reflection density of 1000 reflections/cm or more. However, the potential lowering of the detection limit by use of thin-film waveguides is achievable only if the waveguide material is nonfluorescent and low-loss. Most present evanescent immunosensing technology ("thick" waveguides) utilizes silica glass (SiO2), which is intrinsically nonfluorescent. Only the purest grades of silica, for example UV grade quartz which is rather expensive, lack the additives and impurities that fluoresce (Dierker, et al., 1987).
Further, one cannot simply fabricate silica-on-silica waveguides by depositing SiO2 onto a quartz substrate because there would be no refractive index difference. Instead one must either (1) fabricate a glass waveguide of higher refractive index than the underlying silica substrate, or (2) deposit a silica waveguide onto a transparent substrate of a lower refractive index. Therefore, other materials must be employed.
Thin film waveguides have been described by Sloper et al. ("A planar indium phosphate monomode waveguide evanescent field immunosensor, Sensors and Actuators B1:589-591, 1990) and Zhou et al. ("An evanescent fluorescence biosensor using ion-exchanged buried waveguides and the enhancement of peak fluorescence", Biosensors and Bioelectronics 6:595-607, 1991. However, neither of these devices was capable of achieving detection of analyte concentrations significantly below 10-10 molar. The waveguide structure of Sloper was of the gradient-index type, formed by diffusion of a dopant into the silica base, which results in a drop-off of dopant concentration with distance from the interface. The waveguide of Zhou had only a single "channel" (measurement region).
Therefore, a need exists for an optical structure useful in an evanescent sensing immunoassay, which provides increased levels of propagated TIR light and increased evanescent field interisity, as well as multiple measurement regions. Such an optical structure should desirably be capable of detection of analyte concentrations of 10-13 M and preferably below 1015 M. A need also remains for an immunosensor including such an optical structure, which is sufficiently inexpensive and practical to be produced as a commercial device, and which provides accurate and repeatable results in the hands of non-skilled persons. Still further, a need also remains for an biosensor capable of detecting ions, as opposed to hormones or other biological molecules.

SUlAMARY
The invention comprises a step gradient waveguide, also described as a composite waveguide, useful for performing evanescent sensing assays. The waveguide includes a thick substrate formed of a first optical material of refractive index n, and having a first surface, and a thin film formed of a second optical material having a refractive index n2 which is greater than ni, the thin film being disposed adjacent and in operative contact with the substrate. The optical substrate has a thickness which may be from about 0.3 m up to 10 mm or more, depending on the material used, while the thin film has a thickness which is generally between about 0.3 ftm and about 5 m. Highly preferably, the waveguide thickness is selected to provide for internal propagation in from one to four modes only.
The invention further encompasses a kit comprising the composite waveguide, and at least one specific binding molecule immobilized to said thin film and constructed to bind with specificity an analyte. The kit may be further ' constructed for use in either a competition assay or a sandwich type assay.
The tracer molecule is further constructed to be excited by evanescent light penetrating from the thin film into an adjacent aqueous environment, and to respond thereto by emitting a photodetectable tracer signal.
In a preferred embodiment, the composite waveguide comprises the substrate with a plurality of thin strips of the thin film disposed in parallel array thereon, and the kit further includes a second solution containing a known concentration of analyte in a buffer.
In preferred embodiments of the composite waveguide, coupling means are integrally adapted and in operative contact with the thin film for coupling input light thereinto. One embodiment of coupling means is a grating etched into the substrate on the surface adjacent the thin film. Alternatively, instead of a physical grating a grating-type coupler may be composed of an array of segments of a different refractive index n5 disposed in the substrate in a regular spacing analogous to that of the ridges in a grating. In another alternate embodiment, a relatively thick waveguide coupler is disposed on the planar surface of the thin film waveguide opposite the substrate, near one end of the composite waveguide. The waveguide coupler is dimensioned and constructed of appropriate optical material so as to evanescently couple light propagated by TIR in a thick input waveguide across a thin spacing layer into the thin film waveguide.
In a highly preferred embodiment, the composite waveguide is constructed by vapor deposition of the thin film on the substrate, masking of the thin strips with a resist compound, and etching the thin film to expose the substrate in the unmasked areas. The resist compound is then removed to allow immobilization of the binding molecules to the thin strips.
The invention further embraces apparatus for performing specific binding assays, the apparatus comprising a composite waveguide together with an optical unit having a light source positioned to direct light into the waveguide for propagation by total internal reflectance therein, and detection means oriented to detect light from a region proximal to the optical structure.
The IOW of the invention is capable of detecting analyte concentrations in the femtomolar (10-18 M) range. Such sensitivity is well beyond that achieved by other thin-film evanescent sensors, and also beyond the sensitivity expected solely on the basis of the increased reflection density intensity in the thin-film waveguide.
BRIEF DESCRIPTION OF THE DRAWINGS
In the drawings, which depict the best mode presently known for carrying out the invention:
FIG. 1A is a side cross-sectional view of the composite waveguide of the invention;
FIG. 1B is a top view of a composite waveguide module having a cross-sectional composition according to FIG. 1A;
FIG. 2A is a side view of a composite waveguide having an integral etched grating coupler;
FIG. 2B is a schematic illustration of the embodiment of FIG. 2A showing diffraction of normal incident light beam into a guided mode of the thin film waveguide;
FIG. 3A is a side cross-sectional view of an alternate embodiment of a composite waveguide, having an integral waveguide coupler;
FIG. 3B is a perspective view of the embodiment of FIG. 3A;
FIG. 3C is a schematic illustration of the overlap of the evanescent electromagnetic fields of the input waveguide and the thin film waveguide;
FIG. 4A is a schematic diagram of a top view with partial cutaway of a flow cell incorporating a multichannel composite waveguide;
FIG. 4B is a side cross-section taken along line B-B of FIG. 4A;
FIG. 5 is a schematic diagram of optical apparatus including the flow cell of FIGS. 4A and 4B, for performing binding assays therewith;
FIGS. 6A and 6B are charts of fluorescence measurements made with the apparatus of FIG. 4; and FIG. 7 is a top view of an alternate embodiment of a composite waveguide, having etched sample wells.
DETAILED DESCRIPTION OF THE ILLUSTRATED EMBODIMENTS
A composite waveguide indicated at 100 has a substrate 102 formed of a first optical material of refractive index n1 (FIG. 1A), and a thin-film waveguide layer 103 of a second optical material of refractive index n2, with n2 being greater than nl. The waveguide layer 103 has a plurality of binding molecules 106, each constructed to specifically bind an analyte, immobilized on the upper surface 103A.
The substrate 102 has a thickness D1 which may be from about 0.3 m up to 10 millimeters (mm) or more, while the thin-waveguide 103 has a thickness D2 of between about 0.3 microns ( m) and 5i.cm. If the substrate 102 is relatively thick, perhaps 0.5 mm or more, as shown in FIG. 1A, it may also serve as a mechanical support in addition to having the optical properties necessary for supporting effective waveguiding of light in the thin film waveguide 103. However, in some embodiments it is preferred, generally for easier manufacture, that the substrate 102 be comparatively thin. In such cases an additional support layer (not shown) may be added below the optical substrate 102 (not shown).
In a presently preferred embodiment, the substrate 102 is silicon dioxide, either in the form of a SiO2 thin film laid down by vapor deposition or other techniques as known in the art, or in the form of quartz (here defined as including both natural quartz, and fused silica or other manmade quartz), with a waveguide layer formed of silicon oxynitride (SiZOXNy, or, generally, "SiON"). The term "deposited SiO2" is used hereinafter to distinguish the deposited SiO2 from quartz/fused silica, etc. Silicon dioxide has a refractive index of n = 1.47, and, if sufficiently pure, exhibits very low fluorescence. Silicon oxynitride is mechanically durable, transparent in the visible wavelength range, substantially non-fluorescent, and has a refractive index generally above 1.5 and reaching as high as 2.0 depending on the stoichiometric ratios of 0 to N. In a further presently preferred embodiment, the SiON is Si2O3N.
In another embodiment, the substrate 102 is formed of SiON having a refractive index nl and the waveguide film 103 is formed of SiON having a refractive index n2 > n,. The refractive index of the SiON compositions is controlled by the proportion of nitrogen. That is, the SiON of the substrate will have a lower proportion of nitrogen than the SiON of the waveguide film.
In still another embodiment, the substrate 102 is formed of MgF2 (magnesium fluoride), refractive index n = 1.38, and the waveguide layer 103 is a pure silicon dioxide thin film. Because the MgF substrate is formed by vapor deposition techniques or the like which are tedious and comparatively expensive, in this embodiment the substrate 102 need be only sufficiently thick to ensure efficient propagation of at least one guided mode. At present, the SiON waveguide layer 103 is preferred, as it provides better adherence to the substrate in aqueous solutions than the MgF2 waveguide. However, the MgF2 waveguide/Si02 composite waveguide may be entirely suitable for use with non-aqueous solvents.
In a presently preferred embodiment of a structure including a thin film SiON waveguide, the substrate 102 is a layer of SiO2 deposited on a silicon wafer support. This is preferred because the surface of the substrate 102 is much more uniform and smooth, which in turn produces a much smoother planar surfaces in the deposited SiON film. The increased smoothness of the waveguide surfaces reduces the propagation losses by at least two-fold in comparison with a thin film waveguide deposited on a quartz or fused silica substrate.
In FIG. 1B, the waveguide layer is shown as a plurality of strips 104 which do not extend the full length of the substrate 102. This is an optional configuration of the waveguide layer designed to provide multiple waveguide "channels" for different sample solutions and/or for having different immobilized capture molecules. Further optionally, the composite waveguide 120 may be configured as a plurality of rectangular wells 108A, 108B, 108C, each containing three strips 104.
The strips 104 are separated from one another by a distance D3 of at least about 5 mm. The wells 108A,B,C are separated by walls 110 extending upward from the substrate 102. The device of FIG. 1B may be formed by vapor deposition of the SiON over the entire surface of the substrate 102, masking the regions corresponding to the thin strips 104 with a resist compound, etching the exposed SiON to remove it from the substrate 102, and removing the resist compound. In FIG. 1B, shaded regions 112 represent areas of the waveguide which were not exposed to etchant. The binding molecules 106 are then immobilized to the thin strips 104. This can be accomplished by means known in the art of binding assays.
Alternatively, the wells 108A, 108B, 108C may be configured for flow-cell type operation, as separate flow-through compartments for background, calibration (known analyte concentration), and test sample (unknown) solutions.
High quality composite waveguides made of the material combinations described above can be formed using plasma enhanced chemical vapor deposition (PECVD) to deposit a thin film over an entire surface of the substrate, and then lithographically etching away the thin film except in the desired strips. In the case of SiON, the PECVD is performed with a mixture of nitrogen, nitrous oxide, ammonia and silane gases, and the stoichiometry of SiZOXN,, is varied by changing the respective partial pressures of the above components.
For the lithographic steps, it is presently preferred to use a reactive-iou etch (RIE) process with commercially available photoresist compounds. The areas of the film which are to become the strips or channels are coated with the resist, and the surrounding areas are etched down to the substrate. In the present embodiment, the DUV negative-tone photoresist XP89131 available from Shipley Co. is preferred, and the etchant is a plasma of 02 and CHC13 gases. Positive tone resists such as XP2198 (Shipley Co.), APEX-E from IBM, and CAMP6 from OCG are also suitable. (All of these compositions are proprietary). All of these resist compounds are of the chemical amplification type, comprised substantially of phenolic polymers. While older photo-resists such as PMMA (polymethylmethacrylate) could be used, the chemically amplified DUV resists are considered to produce superior results for the present purposes.
Almost any other etching technique could be readily applied for etching the channels into the waveguide film, including plasma etch, ion milling, or a wet etch.
However, where it is desired to use a substrate-etched grating to increase the coupling efficiency (see FIG. 2A and related description), the RIE process is preferred. This is because the etching process is critical to production of a highly anisotropic grating. RIE etching is believed to provide the best results for the grating, and wet etching would be unsuitable. For waveguides having an etched grating coupler, then, it is convenient to use the RIE process to etch the channels as well.
It is also within contemplation that the thin-film waveguide channels could be produced by a wet lift-off process. In this process, the substrate of the waveguide would be masked to leave the channel regions bare, and the SiON thin film would then be deposited over the whole surface including the masking agent. After deposition of the film, the entire surface would be immersed in a solvent selected to "lift-off" or remove the mask, together with the waveguide film deposited on the mask, while leaving the film deposited on the bare quartz.
All steps of the waveguide fabrication should be performed in an extremely clean environment, preferably a clean-room meeting the standards of at least class 10.

Example 1. SiON composite waveguide fabrication.
Waveguides comprising a 1 m thick film of Si2O3N on SiO2 have been produced as follows. A heated sample holder containing a 10.16 centimeter (4 inch) quartz wafer was placed in a plasma-enhanced chemical vapor deposition (PECVD) reactor. Process gasses flowed from the perimeter of the PECVD vessel, over the sample, and were then pumped out of the vessel through a central port.
During deposition, the PECVD reactor was maintained at 300 C and 1.25 Torr, with 50W of power to a 13.56 MHz generator. The gas mixture consisted essentially of 27 standard cm3 per minute (sccm) silane (SiH4), 500 sccm nitrogen, 200 sccm ammonia, and 1300 sccm nitrous oxide. The respective inlet partial pressures were approximately 17 mTorr, 308 mTorr, 123 mTorr, and 802 mTorr.
Under these conditions, the deposition rate was about 590 A/minute and a 1 m film was produced in about 15 minutes. These silicon oxynitride films had an approximate elemental ratio of Si:O:N = 2:3:1, and a refractive index of about n 1.53 to 1.54.
Next, the Si2O3N films were coated with photoresist, masked, and developed to expose all but nine parallel 1 mm x 65 mm strips of the SiON film separated by 5 mm. The unmasked SiON was etched down to the quartz wafer. The photoresist was stripped, and the etched wafer was cleaved to produce three 23 mm x 69 mm rectangular pieces, where each piece contained three parallel 1 m thick channel waveguides. Only the two outside channels were used in the assays described here.
There are several properties of composite waveguides which can be correlated to their relative suitability for evanescent-sensing biochemical binding assays. These include the ability to withstand the solvent used in the assay, which is generally but not always water; the amount of propagated light lost per unit distance traveled in the waveguide ("propagation loss", expressed as dB per cm; the intrinsic level of fluorescence of the waveguide upon irradiation with light of the excitation wavelength to be used, as measured within the bandwidth of the tracer fluorescence emission; reflections/cm at the waveguide-superstrate interface (N,);
and adequate depth of evanescent penetration (dp).
Example 2. Characterization of SiON wave ug ide.
Scanning electron microscopy was used to examine the shape and thickness of etched channel waveguides. A nominal channel should have optically smooth, rectangular edges and a uniform thickness. For optical characterization of channel waveguides, the beam of the 632.8 nm line from a HeNe laser was coupled into the waveguide using a prism coupler. In a typical experiment, values were determined for waveguide thickness (tWg), refractive index (nwg), internal reflection angle (qwg), reflections/cm at the waveguide-superstrate interface (N), depth of penetration (dP), and propagation loss (dB/cm). The data of Table 1 summarize the properties measured for a typical SiON waveguide constructed by the procedure outlined Table I

Optical characterization of SiON waveguides wave ug ide parameter air superstrate water superstrate twg ( m) 1.31 0.11 N.A.
n,,,g 1.53 0.0 N.A.
propagation loss (dB/cm) 0.76 f 0.09 N.D.
q,,,g (degrees) 82. 67 f 0.47 82. 85 t 0.42 dp (nm) 44.10 f 0.06 68.79 0.29 Nr (reflections/cm) 492.10 71 479 0.29 Each parameter was measured for three different channel waveguides with air as the superstrate. The mean and standard error of these measurements is reported. In addition, the last three parameters (qwg, dP, N) were determined when water was substituted for air as the superstrate. N.D. - not determined.
in Example 1. Also, these waveguides possess a high degree of physical defmition and the desirable features of low propagation loss and minimal intrinsic waveguide fluorescence.

Example 3. Fabrication of MgF2/SiO, composite waveguide.
The SiO2/MgF2 laminates were deposited in situ by electron beam evaporation, using a multipocket electron beam gun in a Balzers BAK760 high-vacuum coater. The deposition chamber was evacuated to 2 Torr, and the silica chips used as mechanical supports on which the laminates were deposited were heated to 200 C. The chamber pressure rose to 3.8 Torr as a result of heating of the silica chips prior to deposition. The source materials, 99.9 % pure MgF2 and 99.999% pure SiO2, were placed in molybdenum- and graphite-lined hearths, respectively, in the rotatable source carousel. Both source materials were brought to the deposition temperature with the shutter closed. First the MgF2 was evaporated with a slowly sweeping 10-kV (kilovolt) electron beam (= 12 mm2 elliptical spot size) followed by similar evaporation of the Si02. The deposition rates of MgF2 and SiOz were controlled at 20 A/s and 10 A/s (angstroms per second), respectively, by means of an oscillating crystal monitor. A 0.36 m film of MgF2 was deposited in 3 minutes, and a 1.0 m film of Si02 was deposited in minutes.
Following construction of the physical portion of the waveguide, a plurality of specific binding molecules, that is, molecules having the property of specifically binding a chosen analyte, are immobilized on the surface of the thin film waveguide channel(s). Such specific binding molecules may be antibodies, receptor molecules, and the like, or fragments thereof that are operative to specifically bind the corresponding analyte. Converse pairs, e.g. such as an antigen for detecting certain antibodies with the analyte being the antibody, are also suitable. Other types of binding molecule/analyte pairs will be apparent to those of ordinary skill, as will means for immobilizing the binding molecule. Presently preferred means for immobilizing the binding molecule are discussed subsequently herein.
A flow cell incorporatiiig a two channel waveguide fabricated and characterized as in Examples 1 and 2 is shown in FIGS. 4A and 4B. A composite waveguide 100 is held between a bottom plate 402 and a top plate 404 spaced by a pair of black silicone rubber o-ring gaskets 406 (FIG. 4B). Each gasket is seated in a corresponding groove 406A milled in the top plate, and forms a longitudinal flow channel 408. The flow channels have respective inlet ports 410A, 412A and outlet ports 410B, 412B. Arrows 409 indicate the direction of flow of liquid through the channel 408. Each flow channel also has a hole 414 for placing a coupling prism 416. The interior of the bottom plate has a window 418 milled therein to support the waveguide and provide a clear view of the waveguide bottom through the silica chip support layer. The aluminum parts of the entire flow cell are desirably flat black, to absorb stray light. Coupling prisms (4 mm wide by 10 mm high truncated 45-45-90 LaSF prisms, np = 1.83, obtained from Karl Lambrecht Co. were fixed in place inside the windows 414 with GTE 118 RTV silicone rubber cement. A series of finger screws (not shown) were used to clamp the top and bottom plates 404, to the substrate 102, and to maintain coupling pressure on the prisms 416.
Polyethylene screw-in tubing connectors, TEFLON microtubing, and disposable syringes served as the sample delivery system (not shown). The sample volume of each channel 408 was 300 L.
Prism coupling the waveguide with aqueous superstrate comprises placing the waveguide on the bottom plate of the flow cell, wetting the waveguide surface opposite the substrate with buffer solution to be used in measurements, and then placing the top plate with the attached prisms over the wetted waveguide. An advantage of the raised-channel waveguide structure is that effective prism coupling is easily accomplished by placing mild pressure on the coupling prisms.
Evanescent binding assays in two model systems were performed with the flow cell of FIGS. 4A, 4B, using a dual-channel assay format similar to that described by Herron et al. in "Fluorescent immunosensors using planar waveguides", SPIE Vol. 1885 (Advances in Fluorescence Sensing Technology), pp.
28-39, 1993, as well as in the copending U.S. Patent Applications Serial Nos.
08/064,608 and 08/263,522.

Example 4. Model assays using thin-film waveg;uide flow cell.
In the first or direct-binding model system, the binding of a biotinylated antibody labelled with a fluorescent tracer to avidin immobilized on the waveguide surface was measured. The particular antibody used was an anti-fluorescein antibody designated 9-40, produced by standard hybridoma methodology. The 9-40 antibody was labelled with the dye Cy-5 650 nm, Xrm,gõX = 667 nm, e.
= 2x105 M-' cm-1) according to the manufacturer's procedure (Biological Detection Systems). The labeling efficiency was approximately four dyes per protein = 5 molecule. An untreated hydrophilic SiON IOW surface was treated for 8 hr with a 0.2 mg/mL solution of avidin (obtained from Sigma) dissolved in phosphate buffered saline (PBS, pH 7.4), to physically adsorb the avidin to the waveguide surface.
A portion of the Cy-5 labeled 9-40 was biotinylated using N-hydroxy-succinimidobiotin (Sigma). Specifically, a 20-fold molar excess of this reagent was added to a 1 mg/mL solution of 9-40 in 0.1 M sodium carbonate/sodium bicarbonate buffer, pH 9 (CBB). This mixture was allowed to react for 2 hr at room temperature, and the product (biotinylated, CY-5-conjugated 9-40) was purified by gel permeation chromatography using a PD-10 column (Pharmacia) equilibrated in PBS.
The second type of assay was an indirect, sandwich type assay in which the binding of free fluorescein-conjugated BSA to immobilized anti-fluorescein antibody (antibody 9-40) was measured using an anti-BSA antibody conjugated with Cy-5 as a tracer. The IOW surfaces with immobilized 9-40 antibody was prepared as follows. Prior to immobilization, 9-40 was acid pre-treated in citrate buffer (pH 3) for 1 hr and reconstituted in PBS at a concentration of 2.3x10-' M (0.03 mg/ml), following the procedure described by Lin et al. (1989). SiON IOWs were treated with 1 % dichlorodimethylsilane (DDS, Sigma), rinsed three times in deionized water, and then immersed in a solution of the acid-pretreated antibody 9-40 for 3 hr at room temperature.
Murine monoclonal anti-bovine serum albumin antibody (anti-BSA; available from Sigma) was labeled with Cy-5 as described for antibody 9-40. Bovine serum albumin (Sigma) was labeled with fluorescein isothiocyanate (FTTC, Sigma). A

fold molar excess of FITC was added to a 1 mg/mi solution of BSA in CBB. This mixture was allowed to react for 1 hr at room temperature, and the conjugate was purified using a PD-10 column equilibrated in PBS. Labeling efficiency was approximately 2 FITC groups per BSA molecule. It should be noted that since the assays were performed with excitation at 632.8 nm, a wavelength which well exceeds the absorption band of FITC (kbs,n,a., = 492 nm), the FITC here serves solely as a hapten for 9-40, and not as a fluorescent tag.
To perform the assays, IOWs coated as described in the preceding paragraphs were mounted in a flow cell like that of FIG. 4, and placed on a rotating goniometer (not shown) in the collection path of a spectrograph 502 associated with a CCD camera 504 (FIG. 5). The goniometer is optional, but useful in an experimental set-up to ascertain the best coupling angle for coupling the laser beam into the waveguide. Once the best coupling angle has been determined and the apparatus standardized, a goniometer is not required to perform the assay. The input light was the red line (6 mW at 632.8 nm) of a HeNe laser (made by Melles Griot), split into dual beams 506, and simultaneously prism coupled into the two channels 104 of the composite waveguide. In this manner, a reference channel and a sample channel were excited.
For detection, the 1 mm wide guided modes in each channel were oriented perpendicular to the entrance slit of the spectrograph. Light emitted from a 1 mm long section of the two parallel streaks was imaged through the window 418 in the bottom plate 402 of the flow cell with a 50 mm f/5.6 camera lens onto the slit of spectrograph 502. The collected light emission was wavelength-dispersed using a single grating monochromator 502 (SPEX 1681c Minimate-2, f/3.9, 300 grooves/mm) centered at 700 nm, and directed onto a thermoelectrically-cooled CCD detector 504 (Photometrics Series 200), producing an image 508 containing reference region 510 and sample region 512. The reference and sample image regions 510, 512 were separately binned and vertically integrated and converted on a MACINTOSH IIx computer to reference spectrum 520 and sample spectrum 522, respectively, of intensity vs. wavelength.
Both assays followed the same general scheme: 1) a capture protein was immobilized to the surface of both the sample and reference channels; 2) aliquots of the hapten-carrier protein conjugate (with fluorescent tracer) were added to the sample channel; and 3) the same concentration of fluorescent tracer (without hapten) was added simultaneously to the reference channel.
Each measurement began by collecting the baseline spectra of PBS buffer in both the sample and reference channels. Starting with the most dilute analyte solution, 1 mL of the sample and reference solutions described in Table 1 were injected into the flow cell. The solutions were incubated with the waveguide surface for 2 min for assay 1 and 5 min for assay 2, followed by collection of sec spectrograph images of both channels. This process was repeated several times until the most concentrated solution was assayed. All CCD images were collected without a wash step; i. e. , with the fluorescent analyte filling the sample and reference channel volume. Reference and sample spectra derived from each CCD
image of each measurement were integrated from 653 nm to 693 nm. Binding curves were constructed from these data by taking the ratio of the integrated intensity of the sample to that of the reference, and then plotting these ratios as a function of bulk analyte concentration. Such radiometric measurements compensated for two effects: nonspecific binding of the tracer, and parasitic excitation of bulk fluorescence.
The results obtained with the two assay schemes are shown in FIGS. 6A-6B.
The results indicate that analyte could be detected at concentrations as low as 3x10-15 M with a two-channel SiZO3N thin film waveguide flow cell. These measurements are at least 3.3x104 times more sensitive than the thin film waveguide immunosensors reported by Sloper et al. or Zhou et al., op cit., and at least times more sensitive than the thick film sensor of Herron et al. 1993, op cit.
The prism coupling scheme described in Example 4 is efficient, simple, and well suited for use in laboratory experiments. However, such a device is relatively expensive, and thus less than ideal for a disposable IOW module for use with a point-of-care optical unit. Further, the prism coupling scheme requires precise pressure on the prisms and precise alignment of the incident laser beam.
An alternate embodiment of a coupling scheme, which avoids these problems, is to provide a diffraction grating etched either into the surface of the waveguide opposite the substrate, or into the substrate adjacent the waveguide. The presently preferred embodiment is that of a substrate-etched grating. As seen in FIG. 2A, the substrate 102 has a grating 200 etched into the surface 102A
which is in contact with the thin film 103. The binding molecules 106 are immobilized down-beam of the grating 200. Optionally, a second grating (not shown) may be etched into the substrate 102, down-beam of the immobilized binding molecules.
Such a second grating is for purposes of optical characterization of the waveguide, and is not required for purposes of the fluorescence binding assay.
The depicted substrate-etched grating is advantageous in that the light can be conveniently in-coupled from the back side of the device, and without interference from macromolecules bound to the waveguide surface. Also, it is convenient and economical from a manufacturing perspective to etch the grating into the substrate prior to deposition and etching of the channel waveguides.
The efficiency of light incoupling by the etched grating will vary according to the spacing, the profile or blaze, and the aspect ratio of the grooves.
These parameters should be optimized according to the wavelength of the input (excitation) light, the index of refraction of the superstrate, and the waveguide thickness, by techniques conventional in the art. In the present case, a grating period D4 of about 0.42 m was computed for coupling of a normally incident 632.8 nm laser beam into the m=0 mode of the thin film SiON waveguide (8 = 83 ; see FIG. 2B).
Still another coupling scheme uses the principle of evanescent penetration to evanescently couple light from a waveguide coupler into the composite waveguide across a precise spacing layer (FIGS. 3A, 3B and 3C). The waveguide coupler is contactingly mounted on the upper surface of the composite waveguide, adjacent the exposed surface of the thin film. Like the prism coupler, the waveguide coupler does not extend into the region to which the specific binding molecules are immobilized.
As seen in FIG. 3A, a waveguide coupler 300 comprises an input waveguide 302 which consists of a comparatively thick (1 mm or greater) layer of material of refractive index n3, and a thin spacing layer 304 of refractive index n4, where n4 is less than both n3 and n2 . The input waveguide has a receiving edge for receiving the input laser light, and a thickness D5 which is sufficient to allow easy end coupling of the input beam into the receiving edge. The spacing layer has a thickness D6 selected to maximize the evanescent coupling of waveguided light from the input waveguide into the thin film of the composite waveguide.
According to coupled mode theory, light propagating in one waveguide can be synchronously coupled into an adjacent waveguide if there is overlap between the evanescent regions of the two waveguides in the low index region separating them (FIG. 3C). The efficiency of coupling depends upon the spacing between the guides, the respective modal propagation constants, and the distance of the interaction. Since the coupling efficiency varies according to the cosine of the distance of interaction, in theory it will reach 100 % at a certain length.
Therefore, the synchronous evanescent coupling device should be quite efficient.
To achieve maximum coupling efficiency, the spacing layer thickness should be adjusted such that K, =(j + 1)7c/2, ( j = 0, 1 , 2...) where K is the coupling constant and depends on the refractive indices n,, n2, n3, n4, the wavelength lambda of the input light, and the respective thicknesses D2, D5 and D6 of the thin film waveguide, the input waveguide, and the spacing layer.
With the thin film waveguide, it should be taken into account that generally, only one or a few of the lowest modes (preferably from j = 0 to about j=10, and no more than j=20) are being propagated in the waveguide. Preferred waveguide couplers are configured to increase the proportion of laser light which is propagated in these lower modes.
For a silicon oxynitride composite waveguide, the spacing layer can be made of SiO2. Desirably, the spacing layer 304 is deposited epitaxially, so that its thickness can be controlled veiy accurately at the time of deposition. The input waveguide 302 is attached on top of the spacing layer, after all epitaxial processing and etching steps have been fniished in the fabrication of the composite waveguide.
The input waveguide is made of a high refractive index material such as rutile, zirconia, or a high-index glass, and may be adhered to the spacing layer 302 by an index-matching cement of refractive index near n3. This assembly would be comparatively inexpensive, and is thus attractive for a disposable, one-time use module.
Preferably, as shown in FIG. 3A, the laser light is focussed into the receiving edge of the input waveguide at an angle 0' which is a degree or two less than the critical angle for TIR at the interface between the input waveguide and the spacing layer. Such angled beam entry has been found to substantially increase the proportion of internally propagated light in the evanescent tail, and thus the evanescent intensity.
Still another embodiment is an IOW with integrally formed sample wells 702, 704, 706 (FIG. 7), each of which may contain multiple thin waveguide channels 104. In a preferred embodiment, walls 700 which enclose the wells are formed by layering a submicron cladding film of the. substrate optical material on the composite waveguide, appropriately masking, and then etching the cladding back down to the SiON waveguide layer. These steps are performed prior to immobilization of the specific binding molecules. Sainple wells thus constructed provide physically defmed regions in which either different antibodies can be immobilized, or to which different solutions, e.g. background or control samples vs.
test samples, can be added. The same technique could be used to form open-ended longitudinal flow channels instead of the box-shaped reservoirs illustrated.
Also, instead of layering the reservoir wall material over the composite waveguide, the same technique could be used to form reservoir or flow channel walls on a separate top plate.
At present it is believed that two additional factors related to the chemistry of immobilization of specific binding molecules to the waveguide surface can affect the sensitivity achieved. These are 1) the relative amount of non-specific protein binding to the thin-film surface (including the walls of any sample wells);
and 2) the analyte/tracer-binding capacity of the immobilized binding molecules.
In a highly preferred embodiment of the multi-channel IOW, the IOW
surface is treated to minimize non-specific binding which otherwise can cause significant overlap of tracer signal between channels. The problem of such overlap worsens rapidly with increasing numbers of channels per IOW. Take as an example a six-channel waveguide divided into three adjacent-channel pairs in three corresponding separate wells, with anti-CK-MB immobilized to one member of each pair and anti-CK-MM immobilized to the other member. The IOW so constructed would permit simultaneous calibrated measurement of binding of 2 different analytes, CK-MB and CK-MM. If the level of non-specific binding is assumed to be about 5 % of the specific binding level, the channel with anti-CK-MB
covalently attached will be contaminated to about 5 % with non-specifically adsorbed anti-CK-MM. Conversely, the channel with anti-CK-MM will be contaminated to about 5 %
with non-specifically adsorbed anti-CK-MB. Thus, with such a 2-analyte IOW
there could be about 10% crosstalk between the channels, already an undesirable contribution to background level. For a 4-analyte IOW (twelve channels), however, the background contributed by such crosstalk would increase to about 30%, which would seriously impair detection sensitivity. Obviously the problem worsens with increasing numbers of different analytes.
For this reason, it is highly desirable that the channels be coated with a coating that reduces non-specific binding to I % or less of specific binding, prior to covalent immobilization of the specific binding molecule.
A method of enhancing the number of available analyte binding sites is to site-specifically immobilize the analyte binding molecules. For antigen-binding fragments such as F(ab')2'S, this can be accomplished by providing a base coating having pendant maleimido groups. The F(ab')2 fragments are then reduced to yield Fab' fragments which contain a free thiol group at their C-terminal end, and this thiol group readily reacts to covalently couple the F(ab')2 fragment to the maleimido group. Desirably, the coating with pendant maleimido groups also provides the inhibition of non-specific binding discussed in the preceding paragraph; this may for example be accomplished by a base coating comprising chains of polymerized hydrophilic residues adhered via a silica-affinic moiety to the waveguide surface, as disclosed by the copending U.S. patent applications S.N. 08/064,608 and 08/263,522. Protocols for achieving both of these methods can be found in the noted copending applications, and in the literature in Herron et al., "Fluorescent immunosensors using planar waveguides", SPIE 1885:28-39, 1993. Preferably, at least 70% of the immobilized analyte binding molecules should be available for binding.
ides.
Example 5. Nucleic acid assays usingthin-film waveguides.
Introduction Nucleic acid probes are gaining increasing acceptance in clinical diagnostics and may one day equal or surpass the importance of immunoassays. Because of their exquisite sensitivity, evanescent wave sensors have the potential of being able to detect DNA hybridization without prior amplification of the analyte. The strategy of this invention is to employ the nucleic acid analog of an immunological sandwich assay. Specifically, an oligonucleotide primer (capture oligonucleotide) complementary to the sequence of interest (analyte) is immobilized to the waveguide using either amine-reactive, thiol-reactive or the (strep)avidin-biotin coupling chemistry (see Herron, J. N., Caldwell, K. D., Christensen, D. A., Dyer, S., Hlady, V., Huang, P., Janatova, V., Wang, H.-K., and Wei, A.-P. "Fluorescent Immunosensors using Planar Waveguides, " SPEE Vol. 1885 (Advances in Fluorescence Sensing Technology) 28-39 (1993).
Next, a fluorescent label such as Cy-5 is coupled to one end of a second oligonucleotide primer (tracer) which is complementary to another sequence on the analyte. To perform the assay, the analyte is first heated if it is double-stranded (to separate the two strands) and then mixed with the tracer oligonucleotide; this mixture is delivered to the sensor where the analyte hybridizes with the capture nucleotide and brings the tracer oligonucleotide into the evanescent field, where it (the tracer) fluoresces.
For detection of nucleic acids, the major technical barrier is the elimination of the amplification step (e.g., polymerase chain reaction), which is required in present-day assays to increase the amount of nucleic acid up to a measurable level.
In theory, evanescent wave sensors possess the intrinsic sensitivity necessary to measure non-amplified DNA. However, there are many technical barriers to achieving this goal including development of (1) immobilization strategies which give very low levels of non-specific binding (perhaps 10- to 100-fold lower than present day technology); (2) effective reagent dispensing strategies that can deliver femtogram quantities of nucleic acid to the sensor surface without losses in a reasonable amount of time (5-10 min); and (3) effective strategies for denaturing double-stranded analytes into the single-stranded form required for hybridization assays.
Materials and Methods Wave ug ides Planar waveguides are employed for conducting fluorescent nucleic acid hybridization assays. Nucleic acid probes are immobilized as described below to either fused silica waveguides (1.0 x 1.0 x 0.1 cm, CO grade, ESCO) or injection molded polystyrene waveguides (1.0 x 1.0 x 0.05 cm, HCP Diagnostics, Salt Lake city).

Nucleic Acid Probes The transcriptional promoter site of the T3 RNA polymerase is employed as a model oligonucleotide sequence in these studies. The T3 promoter is a 20-mer consisting of the following sequence: 5' AATTAACCCTCACTAAAGGG 3'. This oligonucleotide is synthesized and purified by a molecular biology service facility at the LFT-liversity of Utah. When used in DNA hybridization assays, it is immobilized to waveguides (using procedui=es described below) and used as the "capture"
oligonucleotide. It is also used as the soluble inhibitor in competition assays. A
second oligonucleotide sequence, complementary in sequence to the T3 promoter, is synthesized at the same service facility, fluorescently-labeled and used as the soluble "tracer" oligonucleotide in DINTA hybridization assays. To this end, the oligonucleotide is synthesized with a terminal amino group on the 3' end and labeled with Cy-5 (Molecular Detection Systems, Pittsburgh), a red-emitting fluorescent dye.
Immobilization of oligonucleotide probes to silica waveguides Oligonucleotide probes are coupled to silica waveguides using the avidin/biotin coupling chemistry. The general strategy is to immobilize avidin to the waveguides via electrostatic interactions (silica is negatively charged and avidin positively charged at neutral pH) and then complex biotinylated nucleotides to the immobilized avidin. Biotinylated oligonucleotides are prepared by coupling biotinamidocaproate N-hydroxysuccinimide ester (BCHS, Sigma) to a modified oligonucleotide that contained a 5' amino group. The BCHS derivative is used in order to provide a six carbon spacer between the 5' end of the oligonucleotide and the biotin moiety.. The spacer is thought to provide the oligonucleotide with more conformational flexibility in hybridizing with other nucleotides.
Specifically, a 20-fold molar excess of BCHS is added to a 0.1 mg/mL solution of oligonucleotide in 0.1 M sodium carbonate/sodium bicarbonate buffer, pH 9 (CBB). This mixture is allowed to react for 2 hr at room temperature, and the product (biotinylated oligonucleotide) is purified by :reverse phase FPLC chromatography (Pharmacia) using an acetylnitrile/H20 gradient. Avidin is immobilized to silica surfaces by physical adsorption. To this end, clean silica samples are immersed in an avidin solution [3x101 M in phosphate buffered saline (PBS), pH 7.4] for 3 hr. at room temperature and then washed several times in the same buffer to remove unabsorbed avidin. Avidin-coated surfaces are then immersed in a 1.5x10-' M solution of the biotinylated oligonucleotide in PBS for 1 hour at room temperature. Unbound nucleotide is removed by washing in PBS. In some cases, biotinylated poly(ethylene glycol) (biotin-PEG) is coupled to surfaces following the immobilization of biotinylated oligonucleotides. Biotin-PEG is prepared by reaction of N-hydroxysuccinimidobiotin (NHSB) with NH2-PEG-OCH3 (5000 MW, Sigma) for 2 hours in CBB, and purified by dialysis using Spectra/Por dialysis membranes (1000 MW cutoff). Silica surfaces (coated with avidin and oligonucleotides) are immersed in a solution of biotin-PEG (5x10-8 M or 1x10' M) for 1 hour at room temperature. Unbound biotin-PEG is removed by washing in PBS.
Immobilization of oligonucleotide probes to injection molded poly ~~st, r ene waveguides Similar procedures are employed for immobilizing oligonucleotides to polystyrene waveguides, except that strepavidin is used instead of avidin.
Whereas avidin interacts strong with silica substrates due to electrostatic interactions, our experience has shown that it does not adsorb nearly as well to polystyrene surfaces (which are uncharged and hydrophobic in nature). In contrast, strepavidin is the protein of choice for adsorption to polystyrene surfaces. Although functionally equivalent to avidin (i.e., it also contains four biotin binding sites), strepavidin possesses little charge at neutral pH and retains its biotin binding activity upon adsorption.

Total internal reflection fluorescence measurements Total internal reflection fluorescence (TIRF) spectroscopy is an optical technique that is especially well-suited for measuring the concentration of fluorescent molecules attached to a solid surface such as a waveguide. Because water has a lower index of refraction than either silica or polystyrene, a beam of light traveling through a waveguide and striking its edge will either be refracted into the aqueous phase or reflected totally back into the waveguide, depending on the angle of incidence. In the latter case, the incident and reflected beams interfere to produce a standing wave in the waveguide. This wave has a finite electric field amplitude at the edge of the waveguide, but decays exponentially over a distance of 1000 to 2000 angstroms as one moves into the aqueous phase. This decaying electric field is referred to as the evanescent field.
Because the evanescent field is confmed relatively close to the waveguide's surface, it can be used to detect nucleic acid hybridization reactions which occur at the surface. Waveguides are coated with ohgonucleotides as described above and then mounted in the flowceH described in co-pending patent application serial no.
08/064,608. The fluorescence of the Cy-5 labeled tracer oligonucleotide is excited with either the 632.8 run line of a He-Ne laser (Melles Griot) or the 633 nm line of a semiconductor laser (Power Technology, Inc.). The red line is formed into a sheet beam (with a cross-sectional area of ca. 0.2 cm x 3 cm) and coupled simultaneously into the two channels (sample and reference) of the waveguide using a cylindrical coupling lens. The fluorescence emission of Cy-5 is collected using a min f/5.6 camera lens (Nikon) attached to a charge-coupled device (CCD) (either Photometrics Ltd Series 200 or Santa Barbara Instrument Group ST6).
Fluorescence emission is discriminated from Rayleigh light scattering using either a single grating monochromator (SPEX 1681c Minimate-2, f/3.9, 300 grooves/mm) or a 670 nni bandpass interference filter (Omega).

Assay Scheme The use of planar waveguides and TIRF in detection of DNA hybridization is evaluated in two different assays. The first of these (Assay 1) is a direct binding assay in which one strand of a DNA duplex is immobilized to the waveguide (referred to as the "immobilized strand") and the other strand (referred to as the "soluble strand") is labeled with a fluorescent dye. Formation of double-stranded DNA molecules on the waveguide is monitored as a function of the concentration of fluorescently-labeled soluble strand. The second assay (Assay 2) is a competition assay employing immobilized and soluble strands. In addition to fluorescent dye-labeled species of the soluble strands, unlabeled species of the soluble strand are present. Specifically, different concentrations of the unlabeled soluble strand are mixed with a fixed concentration of the labeled soluble strand and each mixture is allowed to react with the immobilized strand on a waveguide sensor. The formation of double- stranded DNA molecules is monitored using TIRF.
Assays are performed using the two- channel flowcell described in U.S. Patent 5,512,492, issued April 30, 1996. A "sample" solution containing the reagents described above is injected into one channel and a "reference"
solution is injected into the other .
The reference solution is used to measure the degree of non-specific binding between the fluorescently-labeled oligonucleotide and the waveguide and also to correct for fluctuations in laser intensity that occurred during the course of an experiment. Each measurement begins by collecting the baseline fluorescence of PBS buffer in both the sample and reference channels. Starting with the most dilute analyte solution, 1 mL
of the sample and reference solutions are injected into the flowcell. The solutions are allowed to incubate with the waveguide surface for 5 minutes, followed by collection of 10 sec fluorescence images of both channels. This process is repeated several times until the most concentrated solution is assayed. All CCD images are collected without a wash step; i.e., with the fluorescent analyte filling the sample and reference channel volume. Binding curves are constructed from these data by taking the ratio of the intensity of the sample to that of the reference, and then plotting these ratios as a function of bulk analyte concentration. Such radiometric measurements compensate for three effects - nonspecific binding of the tracer, excitation of bulk fluorescence by light scattered off the surface of the waveguide and fluctuations in laser intensity.
It will be apparent that details of the composite waveguide can be varied considerably without departing from the concept and scope of the invention.
The claims alone define the scope of the invention as conceived and as described herein.

Claims (20)

What is claimed is:
1. A biosensor for performing specific binding assays, characterized by:
a step gradient waveguide including a planar substrate (102) comprising a first optical material of refractive index n1 and having a surface (102A) disposed adjacent to and in direct contact with a waveguide film (103) including a plurality of discrete regions (104) comprising a second optical material having a refractive index n2 which is greater than n1, and at least one specific binding molecule (106) immobilized to said waveguide file (103), said binding molecule being specific for assaying an analyte.
2. The biosensor of claim 1, wherein said second optical material is selected from the group consisting of: silicon oxynitride and silicon dioxide; and said first optical material is selected from the group consisting of: silicon dioxide, quartz, fused silica, silicon oxynitride, and magnesium fluoride.
3. The biosensor of claim 1, wherein said first and second optical materials are mutually selected from the group of pairwise combinations consisting of:
silicon dioxide/silicon oxynitride; quartz or fused silica/silicon oxynitride; silicon oxynitride of refractive index n1/silicon oxynitride of refractive index n2; and magnesium fluoride/deposited silicon dioxide.
4. The biosensor of claim 3, wherein said waveguide film (103) is constructed to have propagation loses in air of less then 1 dB/cm.
5. The biosensor of claim 2, wherein said planar substrate surface (102A) has a grating (200) formed thereon, said grating (200) being constructed to facilitate coupling of light from an incident excitation beam into said waveguide film (103).
6. The biosensor of claim 1, wherein said waveguide film (103) is coated with a coating which provides a level of nonspecific protein binding which is less than 10%
of the amount of specific binding of said analyte, said coating being selected from the group consisting of: polymethacryloyl polymer, polyethyleneglycol, and avidin;
and wherein said at least one specific binding molecule (106) is immobilized to said waveguide film (103) by binding to said coating.
7. The biosensor of claim 1, wherein the waveguide film (103) has a thickness of 0.1 m to 10 µm.
8. The biosensor of claim 7, wherein said substrate (102) has a thickness of 0.3 µm to 10 mm.
9. The biosensor of claim 8, wherein said waveguide film (103) has a thickness of 0.3 µm to 5 µm.
10. The biosensor of claim 7, wherein said second optical material is selected from the group consisting of: silicon oxynitride and silicon dioxide; and said first optical material is selected from the group consisting of: silicon dioxide, quartz, fused silica, silicon oxynitride, and magnesium fluoride.
11. The biosensor of claim 7, wherein said waveguide film (103) includes a plurality of discrete, parallel strips (104) comprising a plurality of waveguide channels (104).
12. The biosensor of claim 11, comprising a plurality of spaced grooves in said surface (102A) of said first optical material, said spaced grooves being oriented perpendicular to said plurality of waveguide channels (104), and spacing of said spaced grooves serves as a grating (200) facilitating efficient coupling of light from an incident excitation beam into said waveguide film (103).
13. The biosensor of claim 1, wherein said second optical material is silicon oxynitride having a stoichiometric ratio of Si2O3N.
14. The biosensor of claim 1, wherein said step gradient waveguide further includes a waveguide coupler (300).
15. The biosensor of claim 14, wherein the waveguide coupler (300) is contactingly disposed on said waveguide film (103).
16. The biosensor of claim 15, wherein said waveguide coupler (300) comprises:

an input waveguide (302) constructed to receive light through an edge and to propagate the received light by total internal reflection, and a spacing layer (304) interposed between said input waveguide (302) and said waveguide film (103) and having a thickness selected to optimize the evanescent coupling of light from said input waveguide (302) into said waveguide film (103).
17. The biosensor of claim 16, wherein:

said input waveguide (302) comprises an optical material having a refractive index n3, and has a thickness of 0.5 mm to 5 mm; and said spacing layer (304) comprises an optical material having a refractive index n4, wherein n4 < n2 and n4 < n3, and said spacing layer (304) has a thickness selected to optimize the evanescent coupling of light from said input waveguide (302) into said waveguide film (103).
18. The biosensor of claim 16, wherein said input waveguide (302) comprises an optical material selected from the group consisting of: rutile, zirconia, and high-index glass and said spacing layer (304) comprises silicon dioxide and has a thickness of 0.1 µm to 5µm.
19. The biosensor of claim 16, further comprising:

a plurality of sample wells (108A, 108B, 108C), including an outer layer of refractive index n, on said waveguide film (103) defining said sample wells (108A, 108B, 108C).
20. A kit for performing specific binding assays, comprising:

a biosensor according to any of claims 1-19, wherein the at least one specific binding molecule (106) includes a plurality of different species of specific binding molecules (106), and wherein each of said waveguide channels (104) has a species of said plurality of different species immobilized thereto.
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Families Citing this family (207)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5919712A (en) 1993-05-18 1999-07-06 University Of Utah Research Foundation Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays
US5814565A (en) 1995-02-23 1998-09-29 University Of Utah Research Foundation Integrated optic waveguide immunosensor
US5736423A (en) * 1995-11-16 1998-04-07 Advanced Micro Devices, Inc. Method for depositing very thin PECVD SiO2 in 0.5 micron and 0.35 micron technologies
US6611634B2 (en) 1996-03-19 2003-08-26 University Of Utah Research Foundation Lens and associatable flow cell
SE9602545L (en) 1996-06-25 1997-12-26 Michael Mecklenburg Method of discriminating complex biological samples
JP2001504213A (en) * 1996-08-29 2001-03-27 ツェプトゼンス アクチエンゲゼルシャフト Chemical / biochemical optical sensors
CZ297165B6 (en) * 1997-04-21 2006-09-13 Randox Laboratories Ltd. A British Company Of Ardmore Solid state device for performing multi-analyte assays
EP0991777A1 (en) * 1997-06-18 2000-04-12 Ulrich J. Krull Nucleic acid biosensor diagnostics
US6222619B1 (en) 1997-09-18 2001-04-24 University Of Utah Research Foundation Diagnostic device and method
US6395480B1 (en) 1999-02-01 2002-05-28 Signature Bioscience, Inc. Computer program and database structure for detecting molecular binding events
US6485905B2 (en) * 1998-02-02 2002-11-26 Signature Bioscience, Inc. Bio-assay device
US6338968B1 (en) * 1998-02-02 2002-01-15 Signature Bioscience, Inc. Method and apparatus for detecting molecular binding events
US6368795B1 (en) * 1998-02-02 2002-04-09 Signature Bioscience, Inc. Bio-assay device and test system for detecting molecular binding events
US7794946B1 (en) 1998-02-04 2010-09-14 Life Technologies Corporation Microarray and uses therefor
JP3086674B2 (en) * 1998-02-20 2000-09-11 アヴェンティス・リサーチ・ウント・テクノロジーズ・ゲーエムベーハー・ウント・コー・カーゲー Organic substance detection device that enables finger calibration and organic substance monitoring system using the same
GB9810350D0 (en) * 1998-05-14 1998-07-15 Ciba Geigy Ag Organic compounds
US6346376B1 (en) * 1998-06-03 2002-02-12 Centre Suisse D'electronique Et De Mictotechnique Sa Optical sensor unit and procedure for the ultrasensitive detection of chemical or biochemical analytes
US6576478B1 (en) 1998-07-14 2003-06-10 Zyomyx, Inc. Microdevices for high-throughput screening of biomolecules
US6682942B1 (en) 1998-07-14 2004-01-27 Zyomyx, Inc. Microdevices for screening biomolecules
US6406921B1 (en) 1998-07-14 2002-06-18 Zyomyx, Incorporated Protein arrays for high-throughput screening
US6897073B2 (en) 1998-07-14 2005-05-24 Zyomyx, Inc. Non-specific binding resistant protein arrays and methods for making the same
US6188812B1 (en) * 1998-09-01 2001-02-13 Hung Pin Kao Method and apparatus for enhanced evanescent fluorescence and color filtering using a high refractive index thin film coating
US6422066B1 (en) * 1998-10-31 2002-07-23 Yellow Spring Optical Sensor Co. Pll Sensor capsule for CO2 sensor
US6285807B1 (en) * 1998-11-16 2001-09-04 Trustees Of Tufts College Fiber optic sensor for long-term analyte measurements in fluids
US6192168B1 (en) * 1999-04-09 2001-02-20 The United States Of America As Represented By The Secretary Of The Navy Reflectively coated optical waveguide and fluidics cell integration
DE60030978T2 (en) 1999-07-05 2007-06-14 Novartis Ag METHOD FOR USING A SENSOR UNIT
US6771376B2 (en) 1999-07-05 2004-08-03 Novartis Ag Sensor platform, apparatus incorporating the platform, and process using the platform
US20020132371A1 (en) * 1999-09-27 2002-09-19 Kreimer David I. Amplification of analyte detection by substrates having particle structures with receptors
US20040023293A1 (en) * 1999-09-27 2004-02-05 Kreimer David I. Biochips for characterizing biological processes
JP2003510065A (en) * 1999-09-27 2003-03-18 アレイ バイオサイエンス コーポレイション Particle structures with receptors for analyte detection
US20030232388A1 (en) * 1999-09-27 2003-12-18 Kreimer David I. Beads having identifiable Raman markers
US6603548B2 (en) * 1999-12-03 2003-08-05 Sciperio, Inc. Biosensor
US6399295B1 (en) 1999-12-17 2002-06-04 Kimberly-Clark Worldwide, Inc. Use of wicking agent to eliminate wash steps for optical diffraction-based biosensors
GB0000896D0 (en) * 2000-01-14 2000-03-08 Univ Glasgow Improved analytical chip
US6510263B1 (en) * 2000-01-27 2003-01-21 Unaxis Balzers Aktiengesellschaft Waveguide plate and process for its production and microtitre plate
US6728429B1 (en) 2000-02-16 2004-04-27 Biotell, Inc. Optical detection
JP2003531372A (en) * 2000-04-14 2003-10-21 ツェプトゼンス アクチエンゲゼルシャフト Grating waveguide structure and its use for enhancing the excitation field
US7175811B2 (en) 2000-04-28 2007-02-13 Edgelight Biosciences Micro-array evanescent wave fluorescence detection device
US7396675B2 (en) * 2000-06-02 2008-07-08 Bayer Technology Services Gmbh Kit and method for determining a plurality of analytes
US7678539B2 (en) 2000-08-10 2010-03-16 Corning Incorporated Arrays of biological membranes and methods and use thereof
US6977155B2 (en) 2000-08-10 2005-12-20 Corning Incorporated Arrays of biological membranes and methods and use thereof
US6600558B2 (en) * 2000-08-22 2003-07-29 Nippon Telegraph And Telephone Corporation Micro-fluidic cell for optical detection of gases and method for producing same
US7087179B2 (en) * 2000-12-11 2006-08-08 Applied Materials, Inc. Optical integrated circuits (ICs)
US6707548B2 (en) 2001-02-08 2004-03-16 Array Bioscience Corporation Systems and methods for filter based spectrographic analysis
WO2002074899A1 (en) * 2001-03-15 2002-09-26 Array Bioscience Corporation Enhancing surfaces for analyte detection
US6469677B1 (en) 2001-05-30 2002-10-22 Hrl Laboratories, Llc Optical network for actuation of switches in a reconfigurable antenna
US7076092B2 (en) * 2001-06-14 2006-07-11 The United States Of America As Represented By The United States Department Of Energy High-throughput, dual probe biological assays based on single molecule detection
US20040166593A1 (en) * 2001-06-22 2004-08-26 Nolte David D. Adaptive interferometric multi-analyte high-speed biosensor
FR2827386B1 (en) 2001-07-11 2003-10-31 Centre Nat Rech Scient BIOPUCE AND ITS MANUFACTURING METHOD
SE526185C2 (en) * 2001-11-07 2005-07-19 Prolight Diagnostics Ab Method and apparatus for immunoassay
US7102752B2 (en) * 2001-12-11 2006-09-05 Kimberly-Clark Worldwide, Inc. Systems to view and analyze the results from diffraction-based diagnostics
US7029631B2 (en) * 2002-04-19 2006-04-18 Agilent Technologies, Inc. Apparatus for improved light collection
US6670210B2 (en) * 2002-05-01 2003-12-30 Intel Corporation Optical waveguide with layered core and methods of manufacture thereof
US7118855B2 (en) * 2002-05-03 2006-10-10 Kimberly-Clark Worldwide, Inc. Diffraction-based diagnostic devices
US7485453B2 (en) * 2002-05-03 2009-02-03 Kimberly-Clark Worldwide, Inc. Diffraction-based diagnostic devices
US7771922B2 (en) 2002-05-03 2010-08-10 Kimberly-Clark Worldwide, Inc. Biomolecule diagnostic device
US7214530B2 (en) * 2002-05-03 2007-05-08 Kimberly-Clark Worldwide, Inc. Biomolecule diagnostic devices and method for producing biomolecule diagnostic devices
US7091049B2 (en) * 2002-06-26 2006-08-15 Kimberly-Clark Worldwide, Inc. Enhanced diffraction-based biosensor devices
US7169550B2 (en) * 2002-09-26 2007-01-30 Kimberly-Clark Worldwide, Inc. Diffraction-based diagnostic devices
WO2004042429A2 (en) * 2002-10-31 2004-05-21 Luna Innovations, Inc. Fiber-optic flow cell and method relating thereto
US20040086244A1 (en) 2002-11-05 2004-05-06 Zoorob Majd E. Optical waveguide structure
US6985664B2 (en) * 2003-08-01 2006-01-10 Corning Incorporated Substrate index modification for increasing the sensitivity of grating-coupled waveguides
EP3006039B1 (en) 2004-03-02 2021-01-06 Acceleron Pharma Inc. Alk7 polypeptides for use in promoting fat loss
IL161196A0 (en) * 2004-03-04 2005-11-20 Nova Measuring Instr Ltd Polarizer device and method of its manufacture
US7079740B2 (en) * 2004-03-12 2006-07-18 Applied Materials, Inc. Use of amorphous carbon film as a hardmask in the fabrication of optical waveguides
US20050220984A1 (en) * 2004-04-02 2005-10-06 Applied Materials Inc., A Delaware Corporation Method and system for control of processing conditions in plasma processing systems
US7815854B2 (en) 2004-04-30 2010-10-19 Kimberly-Clark Worldwide, Inc. Electroluminescent illumination source for optical detection systems
US7796266B2 (en) 2004-04-30 2010-09-14 Kimberly-Clark Worldwide, Inc. Optical detection system using electromagnetic radiation to detect presence or quantity of analyte
DK2332977T3 (en) 2004-07-23 2016-02-29 Acceleron Pharma Inc ActRII receptor polypeptides
AU2005289414B2 (en) 2004-09-27 2010-12-09 Government Of The United States Of America, Represented By The Secretary Department Of Health And Human Services Nitric oxide-releasing diazeniumdiolated acrylonitrile-based polymers, and compositions, medical devices, and uses thereof
US7209223B1 (en) 2004-11-15 2007-04-24 Luna Innovations Incorporated Optical device for measuring optical properties of a sample and method relating thereto
US7285420B2 (en) * 2004-11-18 2007-10-23 Corning Incorporated System and method for self-referencing a sensor in a micron-sized deep flow chamber
JP4004505B2 (en) * 2005-01-11 2007-11-07 富士フイルム株式会社 Channel member and sensor unit
US7910356B2 (en) * 2005-02-01 2011-03-22 Purdue Research Foundation Multiplexed biological analyzer planar array apparatus and methods
US20070023643A1 (en) * 2005-02-01 2007-02-01 Nolte David D Differentially encoded biological analyzer planar array apparatus and methods
WO2006083917A2 (en) * 2005-02-01 2006-08-10 Purdue Research Foundation Laser scanning interferometric surface metrology
JP4607684B2 (en) * 2005-06-29 2011-01-05 富士フイルム株式会社 Flow path block, sensor unit, and measuring device using total reflection attenuation
US7928079B2 (en) * 2005-10-31 2011-04-19 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Polysaccharide-derived nitric oxide-releasing carbon-bound diazeniumdiolates
CN100427924C (en) * 2005-11-02 2008-10-22 天津科技大学 Self-illuminating biological chip and producing and detecting method thereof
KR20160137665A (en) 2005-11-23 2016-11-30 악셀레론 파마 인코포레이티드 Activin-actrπa antagonists and uses for promoting bone growth
DE102005062377A1 (en) * 2005-12-23 2007-06-28 Bayer Technology Services Gmbh Method for rapid detection of mycotoxins, useful for analysis of food and environmental samples, uses a waveguide that carries binding agents for the toxins
JP4365832B2 (en) 2006-03-07 2009-11-18 株式会社日立製作所 Biochemical analysis cell, biochemical analysis kit and biochemical analysis device
US9423397B2 (en) 2006-03-10 2016-08-23 Indx Lifecare, Inc. Waveguide-based detection system with scanning light source
US9976192B2 (en) 2006-03-10 2018-05-22 Ldip, Llc Waveguide-based detection system with scanning light source
US9528939B2 (en) 2006-03-10 2016-12-27 Indx Lifecare, Inc. Waveguide-based optical scanning systems
US8288157B2 (en) 2007-09-12 2012-10-16 Plc Diagnostics, Inc. Waveguide-based optical scanning systems
US7951583B2 (en) * 2006-03-10 2011-05-31 Plc Diagnostics, Inc. Optical scanning system
US20070259366A1 (en) * 2006-05-03 2007-11-08 Greg Lawrence Direct printing of patterned hydrophobic wells
EP2018541A2 (en) * 2006-05-17 2009-01-28 Siemens Healthcare Diagnostics Inc. Method for signal intensity correction in waveguide sensors
US8207509B2 (en) 2006-09-01 2012-06-26 Pacific Biosciences Of California, Inc. Substrates, systems and methods for analyzing materials
US8471230B2 (en) 2006-09-01 2013-06-25 Pacific Biosciences Of California, Inc. Waveguide substrates and optical systems and methods of use thereof
US20080230605A1 (en) * 2006-11-30 2008-09-25 Brian Weichel Process and apparatus for maintaining data integrity
US7522282B2 (en) * 2006-11-30 2009-04-21 Purdue Research Foundation Molecular interferometric imaging process and apparatus
US20080144899A1 (en) * 2006-11-30 2008-06-19 Manoj Varma Process for extracting periodic features from images by template matching
EP2468291B1 (en) 2006-12-18 2017-11-22 Acceleron Pharma, Inc. Activin-actrii antagonists and uses for increasing red blood cell levels
CN101568823B (en) * 2006-12-21 2013-03-27 皇家飞利浦电子股份有限公司 Aperture biosensor with trenches
WO2008089495A2 (en) * 2007-01-19 2008-07-24 Purdue Research Foundation System with extended range of molecular sensing through integrated multi-modal data acquisition
EP2599495A1 (en) 2007-02-01 2013-06-05 Acceleron Pharma, Inc. Activin-ActRIIa Antagonists and Uses for Treating or Preventing Breast Cancer
TW201803890A (en) 2007-02-02 2018-02-01 艾瑟勒朗法瑪公司 Variants derived from ActRIIB and uses therefor
EA025371B1 (en) 2007-02-09 2016-12-30 Акселерон Фарма Инк. ACTIVIN-ActRIIa ANTAGONISTS AND USE FOR PROMOTING BONE GROWTH IN CANCER PATIENTS
WO2008118934A1 (en) * 2007-03-26 2008-10-02 Purdue Research Foundation Method and apparatus for conjugate quadrature interferometric detection of an immunoassay
CN101607995B (en) 2007-06-15 2013-05-01 厦门大学 Monoclonal antibody or binding activity fragment thereof of H5 subtype avian influenza virus hemagglutinin protein and application thereof
DE102007033124B4 (en) 2007-07-16 2012-12-06 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Device for the optical detection of substances in a liquid or gaseous medium
EP2190201A4 (en) * 2007-09-18 2011-07-06 Panasonic Corp Dispay device, display method and display program
US7960343B2 (en) 2007-09-18 2011-06-14 Acceleron Pharma Inc. Activin-ActRIIa antagonists and uses for decreasing or inhibiting FSH secretion
JP2011503536A (en) * 2007-11-05 2011-01-27 コーニンクレッカ フィリップス エレクトロニクス エヌ ヴィ Microelectronic sensor
GB2461026B (en) 2008-06-16 2011-03-09 Plc Diagnostics Inc System and method for nucleic acids sequencing by phased synthesis
AU2009262968A1 (en) 2008-06-26 2009-12-30 Acceleron Pharma Inc. Antagonists of activin-ActRIIa and uses for increasing red blood cell levels
PT2340031T (en) 2008-08-14 2019-07-25 Acceleron Pharma Inc Gdf traps for use to treat anemia
DE102009029504A1 (en) 2008-09-18 2010-03-25 C. Rob. Hammerstein Gmbh & Co. Kg Motor vehicle seat has seat frame, backrest and hinge fitting arranged between seat frame and backrest, where control cam is arranged on backrest, which is movably connected with hand lever
WO2010099219A2 (en) 2009-02-24 2010-09-02 The Salk Institute For Biological Studies Designer ligands of tgf-beta superfamily
CN107970445B (en) 2009-03-30 2021-09-07 阿塞勒隆制药公司 BMP-ALK3 antagonists and uses for promoting bone growth
KR101766494B1 (en) 2009-04-15 2017-08-08 쓰리엠 이노베이티브 프로퍼티즈 컴파니 Optical film for preventing optical coupling
US9291752B2 (en) 2013-08-19 2016-03-22 3M Innovative Properties Company Retroreflecting optical construction
WO2010120845A2 (en) 2009-04-15 2010-10-21 3M Innovative Properties Company Backlight and display system incorporating same
TWI605276B (en) 2009-04-15 2017-11-11 3M新設資產公司 Optical construction and display system incorporating same
BRPI1006654A2 (en) 2009-04-15 2016-02-10 3M Innovative Properties Co retroreflective optical constructions and optical films
CA2759396A1 (en) 2009-04-29 2010-11-04 Plc Diagnostics Inc. Waveguide-based detection system with scanning light source
BRPI1010587A2 (en) 2009-06-08 2019-04-09 Acceleron Pharma Inc. Methods to Increase Thermogenic Adipocytes
WO2010144150A2 (en) * 2009-06-12 2010-12-16 Pacific Biosciences Of California, Inc. Real-time analytical methods and systems
CN113577291A (en) 2009-08-13 2021-11-02 阿塞勒隆制药公司 Combined use of GDF traps and erythropoietin receptor activators to increase red blood cell levels
US20110070233A1 (en) 2009-09-09 2011-03-24 Acceleron Pharma Inc. Actriib antagonists and dosing and uses thereof
ES2869864T3 (en) 2009-11-03 2021-10-26 Acceleron Pharma Inc Procedures for treating fatty liver disease
JP6267425B2 (en) 2009-11-17 2018-01-24 アクセルロン ファーマ, インコーポレイテッド ACTRIIB protein and its variants and uses thereof for utrophin induction for the treatment of muscular dystrophy
CA2782320A1 (en) 2009-12-02 2011-06-09 Acceleron Pharma Inc. Compositions and methods for increasing serum half-life of fc fusion proteins
EP2933629B1 (en) 2010-02-19 2019-04-10 Pacific Biosciences Of California, Inc. System for measuring analytical reactions comprising a socket for an optode array chip
US8994946B2 (en) 2010-02-19 2015-03-31 Pacific Biosciences Of California, Inc. Integrated analytical system and method
CN102844175B (en) 2010-04-15 2016-08-03 3M创新有限公司 Including optical active areas with without the retroreflective articles of optical active areas
WO2011129832A1 (en) 2010-04-15 2011-10-20 3M Innovative Properties Company Retroreflective articles including optically active areas and optically inactive areas
KR101849889B1 (en) 2010-04-15 2018-04-17 쓰리엠 이노베이티브 프로퍼티즈 캄파니 Retroreflective articles including optically active areas and optically inactive areas
CN102762966B (en) * 2010-04-29 2014-08-06 台湾超微光学股份有限公司 Optomechanical module of micro-spectrometer with conical slit and slit structure thereof
DE202011001569U1 (en) * 2011-01-14 2012-03-01 Berthold Technologies Gmbh & Co. Kg Device for measuring optical properties in microplates
WO2013036829A1 (en) 2011-09-09 2013-03-14 Genentech, Inc Treatment of th17 mediated inflammatory diseases
KR20220075438A (en) 2011-10-17 2022-06-08 악셀레론 파마 인코포레이티드 Methods and compositions for treating ineffective erythropoiesis
US9372308B1 (en) 2012-06-17 2016-06-21 Pacific Biosciences Of California, Inc. Arrays of integrated analytical devices and methods for production
EP3308796B1 (en) 2012-11-02 2021-07-14 Celgene Corporation Activin-actrii antagonists and uses for treating bone and other disorders
EP4123294A1 (en) 2012-12-18 2023-01-25 Pacific Biosciences Of California, Inc. An optical analytical device
US9624540B2 (en) 2013-02-22 2017-04-18 Pacific Biosciences Of California, Inc. Integrated illumination of optical analytical devices
AU2014277045B2 (en) * 2013-06-03 2017-07-13 Ventana Medical Systems, Inc. Fluorescence imaging system for tissue detection
US20150077756A1 (en) * 2013-06-13 2015-03-19 Lumense, Inc. System and method for continuous real-time monitoring of water at contaminated sites
JP6715766B2 (en) 2013-06-21 2020-07-01 ザ ジョンズ ホプキンス ユニバーシティ Virion display array for profiling the function and interaction of human membrane proteins
US20150056199A1 (en) 2013-08-22 2015-02-26 Acceleron Pharma, Inc. Tgf-beta receptor type ii variants and uses thereof
MX2016006455A (en) 2013-11-17 2016-12-09 Quantum-Si Incorporated Active-source-pixel, integrated device for rapid analysis of biological and chemical speciments.
US10018566B2 (en) 2014-02-28 2018-07-10 Ldip, Llc Partially encapsulated waveguide based sensing chips, systems and methods of use
MX2016013635A (en) 2014-04-18 2017-02-02 Acceleron Pharma Inc Methods for increasing red blood cell levels and treating sickle-cell disease.
WO2015192111A1 (en) 2014-06-13 2015-12-17 Acceleron Pharma, Inc. Methods and compositions for treating ulcers
MX2021007934A (en) 2014-08-08 2023-01-17 Quantum Si Inc Integrated device for temporal binning of received photons.
WO2016023011A1 (en) 2014-08-08 2016-02-11 Quantum-Si Incorporated Integrated device with external light source for probing, detecting, and analyzing molecules
CA2957543A1 (en) 2014-08-08 2016-02-11 Quantum-Si Incorporated Optical system and assay chip for probing, detecting and analyzing molecules
EP3186617A4 (en) 2014-08-27 2018-04-25 Pacific Biosciences Of California, Inc. Arrays of integrated analytcal devices
US9400269B2 (en) * 2014-09-22 2016-07-26 The Royal Institution For The Advancement Of Learning/Mcgill University Systems for detecting target chemicals and methods for their preparation and use
MA41052A (en) 2014-10-09 2017-08-15 Celgene Corp TREATMENT OF CARDIOVASCULAR DISEASE USING ACTRII LIGAND TRAPS
WO2016086043A1 (en) 2014-11-24 2016-06-02 Massachusetts Institute Of Technology Methods and apparatus for spectral imaging
TWI730949B (en) 2014-12-03 2021-06-21 美商西建公司 Activin-actrii antagonists and uses for treating anemia
MA41119A (en) 2014-12-03 2017-10-10 Acceleron Pharma Inc METHODS OF TREATMENT OF MYELODYSPLASIC SYNDROMES AND SIDEROBLASTIC ANEMIA
US10302972B2 (en) * 2015-01-23 2019-05-28 Pacific Biosciences Of California, Inc. Waveguide transmission
US11181479B2 (en) 2015-02-27 2021-11-23 Ldip, Llc Waveguide-based detection system with scanning light source
US10487356B2 (en) 2015-03-16 2019-11-26 Pacific Biosciences Of California, Inc. Integrated devices and systems for free-space optical coupling
WO2016164501A1 (en) 2015-04-06 2016-10-13 Acceleron Pharma Inc. Single-arm type i and type ii receptor fusion proteins and uses thereof
MA41919A (en) 2015-04-06 2018-02-13 Acceleron Pharma Inc ALK4 HETEROMULTIMERS: ACTRIIB AND THEIR USES
EP3828199A1 (en) 2015-04-06 2021-06-02 Acceleron Pharma Inc. Alk7: actriib heteromultimers and uses thereof
US11466316B2 (en) 2015-05-20 2022-10-11 Quantum-Si Incorporated Pulsed laser and bioanalytic system
US10605730B2 (en) 2015-05-20 2020-03-31 Quantum-Si Incorporated Optical sources for fluorescent lifetime analysis
EP3308204A4 (en) 2015-06-12 2019-03-13 Pacific Biosciences of California, Inc. Integrated target waveguide devices and systems for optical coupling
US9851290B2 (en) * 2015-06-22 2017-12-26 Sharp Laboratories Of America, Inc. Particle detector for particulate matter accumulated on a surface
WO2017019482A1 (en) 2015-07-24 2017-02-02 Massachusetts Institute Of Technology Apparatus, systems, and methods for biomedical imaging and stimulation
EP4218792A1 (en) 2015-08-04 2023-08-02 Acceleron Pharma Inc. Composition for treating myeloproliferative disorders
US10550170B2 (en) 2015-11-23 2020-02-04 Acceleron Pharma Inc. Methods for treating vascular eye disorders with actrii antagonists
US10006809B2 (en) 2016-02-10 2018-06-26 Massachusetts Institute Of Technology Apparatus, systems, and methods for on-chip spectroscopy using optical switches
WO2017143129A1 (en) 2016-02-17 2017-08-24 Tesseract Health, Inc. Sensor and device for lifetime imaging and detection applications
US10151879B2 (en) * 2016-05-19 2018-12-11 Raytheon Bbn Technologies Corporation Photonic device for ultraviolet and visible wavelength range
EP3464337A4 (en) 2016-05-31 2019-12-18 Mogam Institute for Biomedical Research Ab6 family designer ligands of tgf-beta superfamily
WO2018013243A1 (en) 2016-06-01 2018-01-18 Quantum-Si Incorporated Integrated device for detecting and analyzing molecules
US10722558B2 (en) 2016-07-15 2020-07-28 Acceleron Pharma Inc. Compositions and methods for treating pulmonary hypertension
MA45811A (en) 2016-07-27 2019-06-05 Acceleron Pharma Inc METHODS AND COMPOSITIONS OF TREATMENT OF DISEASE.
CA3036104A1 (en) 2016-09-15 2018-03-22 Acceleron Pharma Inc. Twisted gastrulation polypeptides and uses thereof
CN110198743B (en) 2016-10-05 2023-07-18 艾科赛扬制药股份有限公司 Compositions and methods for treating kidney disease
WO2018112170A1 (en) 2016-12-16 2018-06-21 Quantum-Si Incorporated Compact beam shaping and steering assembly
KR102330080B1 (en) 2016-12-16 2021-11-25 퀀텀-에스아이 인코포레이티드 Compact Mode Synchronous Laser Module
CN110168732B (en) 2016-12-22 2024-03-08 宽腾矽公司 Integrated photodetector with direct combined pixels
JP7267664B2 (en) 2017-05-04 2023-05-02 アクセルロン ファーマ インコーポレイテッド TGFbeta receptor type II fusion protein and uses thereof
KR20200028474A (en) * 2017-07-24 2020-03-16 퀀텀-에스아이 인코포레이티드 Optical rejection photon structures
WO2019032735A1 (en) 2017-08-08 2019-02-14 Massachusetts Institute Of Technology Miniaturized fourier-transform raman spectrometer systems and methods
CN111867729B (en) 2018-01-08 2022-06-10 宽腾矽公司 System and method for electrokinetic loading of sub-micron reaction chambers
WO2019147904A1 (en) 2018-01-26 2019-08-01 Quantum-Si Incorporated Machine learning enabled pulse and base calling for sequencing devices
WO2019190740A1 (en) * 2018-03-29 2019-10-03 Illumina, Inc. Illumination for fluorescence imaging using objective lens
CA3098712A1 (en) 2018-05-03 2019-11-07 Quantum-Si Incorporated Characterizing an optical element
JP7317050B2 (en) 2018-05-14 2023-07-28 クアンタム-エスアイ インコーポレイテッド Systems and methods for integrating statistical models of different data modalities
KR20210021018A (en) 2018-06-15 2021-02-24 퀀텀-에스아이 인코포레이티드 Data acquisition control for advanced analysis instruments with pulsed optical sources
EP3811610A1 (en) 2018-06-22 2021-04-28 Quantum-Si Incorporated Integrated photodetector with charge storage bin of varied detection time
US11041759B2 (en) 2018-06-28 2021-06-22 Massachusetts Institute Of Technology Systems and methods for Raman spectroscopy
EP3837532A1 (en) 2018-08-17 2021-06-23 University of Rochester Optical biosensor comprising disposable diagnostic membrane and permanent photonic sensing device
BR112021003074A2 (en) 2018-08-29 2021-05-11 Quantum-Si Incorporated sample well fabrication techniques and structures for integrated sensor devices
CN113424047A (en) 2019-01-03 2021-09-21 宽腾矽公司 Optical waveguide and coupler for delivering light to an array of photonic elements
WO2020167370A1 (en) 2019-02-11 2020-08-20 Massachusetts Institute Of Technology High-performance on-chip spectrometers and spectrum analyzers
BR112021025186A2 (en) 2019-06-14 2022-04-12 Quantum Si Inc Sliced grid coupler with increased beam alignment sensitivity
WO2020257445A1 (en) 2019-06-19 2020-12-24 Quantum-Si Incorporated Optical nanostructure rejecter for an integrated device and related methods
KR20220025853A (en) 2019-06-28 2022-03-03 퀀텀-에스아이 인코포레이티드 Elimination of optical and electrical secondary paths
CN114514420A (en) 2019-08-08 2022-05-17 宽腾矽公司 Increased emission collection efficiency in integrated optical devices
KR20220115630A (en) 2019-12-09 2022-08-17 클라우디오 올리베이라 에갈론 Side Illumination Systems and Methods of Waveguides
JP2023510577A (en) 2020-01-14 2023-03-14 クアンタム-エスアイ インコーポレイテッド Integrated sensor for lifetime characterization
IL294732A (en) 2020-01-14 2022-09-01 Quantum Si Inc Sensor for lifetime plus spectral characterization
EP4111178A1 (en) 2020-03-02 2023-01-04 Quantum-si Incorporated Integrated sensor for multi-dimensional signal analysis
US11573180B2 (en) 2020-04-08 2023-02-07 Quantum-Si Incorporated Integrated sensor with reduced skew
US20210349018A1 (en) * 2020-05-07 2021-11-11 Hand Held Products, Inc. Apparatuses, systems, and methods for sample testing
US20220128402A1 (en) 2020-10-22 2022-04-28 Quantum-Si Incorporated Integrated circuit with sequentially-coupled charge storage and associated techniques
US11846574B2 (en) 2020-10-29 2023-12-19 Hand Held Products, Inc. Apparatuses, systems, and methods for sample capture and extraction

Family Cites Families (89)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4264766A (en) * 1877-09-19 1981-04-28 Hoffmann-La Roche Inc. Immunological diagnostic reagents
US3449037A (en) * 1966-03-14 1969-06-10 American Optical Corp Fiber optical image-enhancing devices,systems,and the like
US3934061A (en) * 1972-03-30 1976-01-20 Corning Glass Works Method of forming planar optical waveguides
US4166105A (en) * 1973-07-30 1979-08-28 Block Engineering, Inc. Dye tagged reagent
US3939350A (en) * 1974-04-29 1976-02-17 Board Of Trustees Of The Leland Stanford Junior University Fluorescent immunoassay employing total reflection for activation
USRE34394E (en) * 1978-01-23 1993-09-28 Baxter Diagnostics Inc. Method and composition for double receptor, specific binding assays
JPS5510590A (en) * 1978-05-04 1980-01-25 Wellcome Found Enzyme immunity quantity analysis
US4399099A (en) * 1979-09-20 1983-08-16 Buckles Richard G Optical fiber apparatus for quantitative analysis
JPS58501481A (en) * 1981-09-18 1983-09-01 プルーテック リミティド Method and device for measuring test object in solution using optical waveguide
USRE33064E (en) * 1981-09-18 1989-09-19 Prutec Limited Method for the determination of species in solution with an optical wave-guide
US4450231A (en) * 1982-03-31 1984-05-22 Biostar Medical Products, Inc. Immunoassay for determination of immune complexes with polymer-coated plastic base
DE3344019C2 (en) * 1983-12-06 1995-05-04 Max Planck Gesellschaft Device for optically measuring the concentration of a component contained in a sample
US4775637A (en) * 1984-12-10 1988-10-04 Purtec Limited An immunoassay apparatus having at least two waveguides and method for its use
EP0184600B1 (en) * 1984-12-10 1990-03-14 Prutec Limited Method for optically ascertaining parameters of species in a liquid analyte
GB8509491D0 (en) * 1985-04-12 1985-05-15 Plessey Co Plc Optic waveguide biosensors
WO1986007149A1 (en) 1985-05-29 1986-12-04 Kurt Tiefenthaler Optical sensor for selectively determining the presence of substances and the variation of the refraction index in the measured substances
US4945245A (en) * 1986-01-14 1990-07-31 Levin Herman W Evanescent wave background fluorescence/absorbance detection
DE3782394T2 (en) * 1986-03-28 1993-05-19 Toray Industries IMMOBILIZED, PHYSIOLOGICALLY ACTIVE MATERIAL.
US4935346A (en) * 1986-08-13 1990-06-19 Lifescan, Inc. Minimum procedure system for the determination of analytes
US4849340A (en) * 1987-04-03 1989-07-18 Cardiovascular Diagnostics, Inc. Reaction system element and method for performing prothrombin time assay
GB8714503D0 (en) 1987-06-20 1987-07-22 Pa Consulting Services Detector
US5006333A (en) * 1987-08-03 1991-04-09 Ddi Pharmaceuticals, Inc. Conjugates of superoxide dismutase coupled to high molecular weight polyalkylene glycols
AU604364B2 (en) * 1987-08-13 1990-12-13 Dow Chemical Company, The Sulfur dioxide removal from gas streams using hydroxyalkyl substituted piperazinones
US4909990A (en) * 1987-09-02 1990-03-20 Myron J. Block Immunoassay apparatus
BR8807884A (en) * 1988-02-14 1990-11-13 Walter Lukosz INTEGRATED OPTICAL INTERFERENCE PROCESS
US4913519A (en) * 1988-03-04 1990-04-03 Fiberchem Inc. Optical sensor for the detection of ice formation and other chemical species
US4866681A (en) * 1988-03-09 1989-09-12 Mine Safety Appliances Company Photo-acoustic detector
GB8807486D0 (en) * 1988-03-29 1988-05-05 Ares Serono Res & Dev Ltd Waveguide sensor
US4893894A (en) * 1988-04-29 1990-01-16 Mine Safety Appliances Company Evanescent sensor
EP0341927B1 (en) * 1988-05-10 1993-07-14 AMERSHAM INTERNATIONAL plc Biological sensors
DE3820171A1 (en) * 1988-06-14 1989-12-21 Messerschmitt Boelkow Blohm WAVE GUIDE / DETECTOR COMBINATION
GB8817710D0 (en) * 1988-07-25 1988-09-01 Ares Serono Res & Dev Ltd Method of assay
US5478755A (en) * 1988-07-25 1995-12-26 Ares Serono Research & Development Ltd. Long range surface plasma resonance immunoassay
US5340722A (en) * 1988-08-24 1994-08-23 Avl Medical Instruments Ag Method for the determination of the concentration of an enzyme substrate and a sensor for carrying out the method
US4940328A (en) * 1988-11-04 1990-07-10 Georgia Tech Research Corporation Optical sensing apparatus and method
SE462408B (en) * 1988-11-10 1990-06-18 Pharmacia Ab OPTICAL BIOSENSOR SYSTEM USING SURFACE MONITORING RESONSE FOR THE DETECTION OF A SPECIFIC BIOMOLIC CYCLE, TO CALIBRATE THE SENSOR DEVICE AND TO CORRECT FOUND BASELINE OPERATION IN THE SYSTEM
GB8827853D0 (en) * 1988-11-29 1988-12-29 Ares Serono Res & Dev Ltd Sensor for optical assay
JP2635732B2 (en) * 1988-12-01 1997-07-30 古河電気工業株式会社 Optical fiber sensing method
US4892383A (en) * 1989-02-17 1990-01-09 Fiberchem Inc. Reservoir fiber optic chemical sensors
US5006716A (en) * 1989-02-22 1991-04-09 Research Corporation Technologies, Inc. Method and system for directional, enhanced fluorescence from molecular layers
US5401469A (en) * 1989-04-19 1995-03-28 Ibiden Co., Ltd. Plastic optical biomaterials assay device
DE59001976D1 (en) * 1989-05-01 1993-08-19 Wolfram Bohnenkamp REFLECTION FLUORIMETER.
US5266486A (en) * 1989-05-12 1993-11-30 Nvl Photronics Corporation Method and apparatus for detecting biological activities in a specimen
GB8911462D0 (en) * 1989-05-18 1989-07-05 Ares Serono Res & Dev Ltd Devices for use in chemical test procedures
US5268305A (en) * 1989-06-15 1993-12-07 Biocircuits Corporation Multi-optical detection system
ATE98773T1 (en) * 1989-06-22 1994-01-15 Ars Holding 89 Nv METHODS OF OPTICAL ANALYSIS.
CA2024548C (en) * 1989-09-05 2002-05-28 David Issachar Analyte specific chemical sensor
JPH03122553A (en) * 1989-10-04 1991-05-24 Olympus Optical Co Ltd Photosensor
US5274721A (en) * 1992-03-05 1993-12-28 American Sigma, Inc. Fiber optic system and method
US5082629A (en) * 1989-12-29 1992-01-21 The Board Of The University Of Washington Thin-film spectroscopic sensor
DE69017471D1 (en) * 1990-02-22 1995-04-06 Univ Mcgill SOLIDS PHASE INTERFEROMETRIC IMMUNOTEST SYSTEM.
WO1991013339A1 (en) * 1990-03-02 1991-09-05 Fisons Plc Sample cell for use in chemical or biochemical assays
US5224188A (en) * 1990-04-20 1993-06-29 Hughes Aircraft Company Eccentric core optical fiber
DE4023671A1 (en) * 1990-07-25 1992-01-30 Boehringer Mannheim Gmbh Non-ionic block copolymers made of propylene oxide and ethylene oxide
US5202230A (en) * 1990-09-07 1993-04-13 Kamentsky Louis A Methods of detecting cut cells in a tissue section
US5377008A (en) * 1990-09-20 1994-12-27 Battelle Memorial Institute Integrated optical compensating refractometer apparatus
US5173747A (en) * 1990-09-20 1992-12-22 Battelle Memorial Institute Integrated optical directional-coupling refractometer apparatus
US5212099A (en) * 1991-01-18 1993-05-18 Eastman Kodak Company Method and apparatus for optically measuring concentration of an analyte
US5192510A (en) * 1991-01-30 1993-03-09 E. I. Du Pont De Nemours And Company Apparatus for performing fluorescent assays which separates bulk and evanescent fluorescence
US5156976A (en) * 1991-06-07 1992-10-20 Ciba Corning Diagnostics Corp. Evanescent wave sensor shell and apparatus
US5340715A (en) 1991-06-07 1994-08-23 Ciba Corning Diagnostics Corp. Multiple surface evanescent wave sensor with a reference
CA2069537A1 (en) * 1991-06-07 1992-12-08 Thomas A. Cook Multiple output referencing system for evanescent wave sensor
US5262638A (en) * 1991-09-16 1993-11-16 The United States Of America As Represented By The United States National Aeronautics And Space Administration Optical fibers and fluorosensors having improved power efficiency and methods of producing same
US5249251A (en) * 1991-09-16 1993-09-28 The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration Optical fiber sensor having an active core
WO1993010226A1 (en) * 1991-11-19 1993-05-27 North Carolina State University Immunodiagnostic assay using liposomes carrying labels thereof on outer liposome surface
US5846708A (en) * 1991-11-19 1998-12-08 Massachusetts Institiute Of Technology Optical and electrical methods and apparatus for molecule detection
JP3107649B2 (en) * 1991-12-20 2000-11-13 イビデン株式会社 Fluorescent immunoassay
US5354574A (en) * 1992-06-23 1994-10-11 Ibiden Co., Ltd. Method for producing optical fiber having formyl groups on core surface thereof
US5327225A (en) * 1993-01-28 1994-07-05 The Center For Innovative Technology Surface plasmon resonance sensor
US5343550A (en) * 1993-02-25 1994-08-30 The United States Of America As Represented By The United States National Aeronautics And Space Administration Transversely polarized source cladding for an optical fiber
US5399866A (en) * 1993-03-24 1995-03-21 General Electric Company Optical system for detection of signal in fluorescent immunoassay
US5512492A (en) * 1993-05-18 1996-04-30 University Of Utah Research Foundation Waveguide immunosensor with coating chemistry providing enhanced sensitivity
JP3426602B2 (en) * 1993-05-18 2003-07-14 ユニヴァーシティ オブ ユタ リサーチ ファンデーション Apparatus and method for multi-analyte homogeneous fluorescence immunoassay
US5677196A (en) * 1993-05-18 1997-10-14 University Of Utah Research Foundation Apparatus and methods for multi-analyte homogeneous fluoro-immunoassays
US5552272A (en) * 1993-06-10 1996-09-03 Biostar, Inc. Detection of an analyte by fluorescence using a thin film optical device
US5413939A (en) * 1993-06-29 1995-05-09 First Medical, Inc. Solid-phase binding assay system for interferometrically measuring analytes bound to an active receptor
GB9314991D0 (en) 1993-07-20 1993-09-01 Sandoz Ltd Mechanical device
US5416579A (en) * 1993-07-23 1995-05-16 Nova Chem Bv Method for determining concentration in a solution using attenuated total reflectance spectrometry
US5494798A (en) * 1993-12-09 1996-02-27 Gerdt; David W. Fiber optic evanscent wave sensor for immunoassay
US5432096A (en) * 1993-12-20 1995-07-11 Cetac Technologies Inc. Simultaneous multiple, single wavelength electromagnetic wave energy absorbtion detection and quantifying spectrophotometric system, and method of use
US5437840A (en) * 1994-04-15 1995-08-01 Hewlett-Packard Company Apparatus for intracavity sensing of macroscopic properties of chemicals
US5538850A (en) * 1994-04-15 1996-07-23 Hewlett-Packard Company Apparatus and method for intracavity sensing of microscopic properties of chemicals
DE69505370T2 (en) * 1994-05-27 1999-04-01 Novartis Ag METHOD AND SENSOR FOR DETECTING DECAYING EXCITED LUMINESCENCE
US5577137A (en) * 1995-02-22 1996-11-19 American Research Corporation Of Virginia Optical chemical sensor and method using same employing a multiplicity of fluorophores contained in the free volume of a polymeric optical waveguide or in pores of a ceramic waveguide
US5814565A (en) * 1995-02-23 1998-09-29 University Of Utah Research Foundation Integrated optic waveguide immunosensor
US5492674A (en) * 1995-03-17 1996-02-20 Boehringer Mannheim Corporation Evanescent wave immunoassay system
US5589136A (en) * 1995-06-20 1996-12-31 Regents Of The University Of California Silicon-based sleeve devices for chemical reactions
US5832165A (en) * 1996-08-28 1998-11-03 University Of Utah Research Foundation Composite waveguide for solid phase binding assays
NL1008166C2 (en) 1998-01-30 1999-08-02 Ambaflex B V Transporter.

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JPH11500826A (en) 1999-01-19
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US5961924A (en) 1999-10-05
WO1996026432A1 (en) 1996-08-29
AU4997696A (en) 1996-09-11
US20020034457A1 (en) 2002-03-21
CA2213694A1 (en) 1996-08-29
AU693666B2 (en) 1998-07-02
US7537734B2 (en) 2009-05-26
EP0812416A1 (en) 1997-12-17
EP0812416A4 (en) 1999-04-07

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