Abstract: Improvements in non-invasive detection methods for glucose and other constituents of interest in a sample have been developed. The apparatus and methods of the invention provide an analog of color perception of human vision, preferably in the near infrared region, replacing spectrophotometers and narrow band sources used in other non-invasive near infrared detection methods. A plurality of detector units are used, each covering a broad and overlapping region of the detected spectrum, paralleling color perception and colorimetry. The improvements are primarily concerned with improving the signal-to-background (or noise) ratio such that the data stream is improved. These improvements use congruent sampling, comparison of different data streams from different sample portions or filter sets, using an interrogation system with sufficient speed to allow testing of arterial blood, and using a filter with a spectral structure. In some circumstances, a neural net is used for analysis, allowing the system to learn.

Abstract: A new non-invasive non-spectrophotometric method for measuring the blood concentration of analytes such as glucose has been developed. The apparatus and methods of the invention exploit analogies with colorimetry and color perception to extract concentration measurements from the global structure of the intensity versus wavelength absorbance or transmission profile. A plurality of broad spectrum filters transmit distinguishably coded beams of radiation in overlapping portions of the spectrum to the sample. Radiation reflected or transmitted by the sample is detected and decoded. LED's may be used instead of the broad spectrum radiation generating device and the filters. Further, a scanning interferometer can be used as the illuminating and coding device. In a preferred mode, congruent illumination is utilized. The coded signals are analyzed by analogy to colorimetry and visual processing and can be converted into concentration measurements.

Abstract: An assay for polynucleotides employing total internal reflection of excitation radiation at a coating bonded to the surface of an optically conductive glass cell. The coating initially includes single-stranded polynucleotides coupled to individual attachment sites on the surface of the cell, such polynucleotides being complementary, at least in part to the single-stranded form of the polynucleotide that is being assayed. Each molecule of the coupled polynucleotide is connected to the cell surface through a spacer connected to an irreversibly conjugated polyadenine/polythymidine sequence at one end of the coupled polynucleotide.When the surface of the coated cell is contacted with a sample that contains single-stranded polynucleotide complementary to the bound polynucleotide, renaturation will occur, forming a double-stranded form of the polynucleotide of interest.

Abstract: An assay apparatus employing total internal reflection of excitation radiation at the interface between a replaceable optically conductive rod or fiber and a surrounding liquid phase of lower index of refraction. Immobilized on the surface of the fiber is a component of a complex formed in an immunological-type specific reaction. A fluorophore that can be excited into fluorescence by the excitation radiation is attached to another component of the complex. The rod is coaxially mounted within a tube that is sized with respect to said rod so that a fluid sample may be introduced into said tube.The rod and tubing are supported in a mounting assembly that is attachable to an optical assembly for transmitting excitation radiation into the proximal end of the rod and receiving fluorescent radiation emitted from the proximal end of the rod. Included in the apparatus is a mounting assembly for centering the rod within tube and for biasing the rod in a first direction against an annular seat.

Abstract: A method and apparatus for fluorescent immunoassay utilizes total internal reflection at the interface between a solid phase and a liquid phase of lower index of refraction to produce an evanescent wave in the liquid phase. The solid phase is arranged and illuminated so as to provide multiple total internal reflections at the interface. In a preferred embodiment, the solid phase is in the form of an optical fiber to which is immobilized a component of the complex formed in the immunochemical reaction. A fluorophore is attached to another component of the complex. The fluorescent labelled component may be either the complement to or an analog of the immobilized component, depending on whether competitive or sandwich assays are to be performed. In the case of competitive assays, the labelled component is preferably pre-loaded to the immobilized component in a controlled concentration. The fiber (and the attached constituent of the assay) is immersed in the liquid phase sample.

Abstract: An assay apparatus and methods employing total internal fluorescence to define a thin observed volume of a multi-phase suspension from which the suspended phase, because of its size and shape, is substantially excluded. Observations of the fluorescence of a known quantity of fluorescent material introduced into a known volume of the sample containing the observed volume permits quantitation of the volumes of the suspended and suspending phases.

Abstract: Fluorescent immunoassay apparatus and method employing a disposable consisting, in a preferred embodiment, of a short length of precise diameter capillary tubing having an axially disposed optical fiber to which is immobilized a monolayer of a component of the antibody-antigen complex (e.g., an antibody), an inert diluent, and a preload of a known amount of tagged complement to the immobilized component (e.g., a fluorescent-tagged antigen). The dimensions of the fiber and the capillary tubing are chosen so as to allow the tube to fill itself by capillary action once an end of the tube is immersed into the sample. Precise timing and ballistic measurements may, if desired, be avoided by insuring the incubation time is larger than the diffusion time necessary to scavenge the volume between the fiber and the capillary tubing. Fluorimetric measurement is made by total reflection fluorescence techniques.