Abstract: A method of immunologically staining a formalin-fixed tissue preparation, which comprises (a) subjecting a formalin-fixed tissue preparation to microwave energy while the tissue preparation is submersed in water for a time sufficient to increase immunostaining efficiency; (b) removing the tissue preparation from the water and cooling; and (c) contacting the tissue preparation with an immunological staining reagent.

Abstract: An immunoassay method for the detection or quantitation of an analyte suspected of being in a solution comprising: (a) combining said specimen, a first binding component, insoluble particles, and second binding component labelled with a signal generating material in a solid phase retention and separation apparatus having a sufficient pore size such that said particles are trapped within said filter yet permitting rapid passage of fluid therethrough in such a manner that an immunological reaction occurs if analyte is present in said specimen, resulting in the formation of an immunocomplex of insolubilized first binding component:analyte:second labelled binding component on or within said filter means; (b) separating bound from unbound material; and (c) determining the presence and/or amount of signal produced which is correlative with the amount of analyte present in the solution.

Abstract: A monoclonal or polyclonal antibody having a specific binding property to an antigenic site on the human sperm acrosome, which is produced by a hybridoma obtained by fusion between an antibody-producing mammalian (except human) cell immunized with the antigenic site on the human sperm acrosome and a cell having permanent proliferation potency.

Abstract: Genotoxic chemicals are an existing wide-spread health hazard to the human population. Advances in genetic toxicology testing have made it possible to assay potential mutagens, carcinogens, teratogens and clastogens in the environment. The mouse micronucleus assay provides an example of an excellent test for genetic damage to cells. When chromosome breaks occur in the blood stem cell population, the damaged piece of chromosome remains behind as a micronucleus in the normally DNA deficient red blood cells. However, currently available manual micronucleus assays are costly, time consuming, and labor intensive. In addition, the statistics are often marginal since the number of micronucleii (MNs) in 1000 polychromatic cells are scored manually, yielding limited amounts of data. This invention discloses the means for assaying the change in micronucleated cells by high speed flow cytometry.

Abstract: A method of monitoring the aggregation of cells in, for example, an immuno-agglutination assay, comprises promoting agglutination sonically in a capillary and inverting the capillary to cause agglutinated particles to settle at a meniscus. The granular appearance of agglutinated cells can be distinguished visually from the smooth distribution of non-aggregated cells.

Abstract: A diagnostic kit for detecting the presence of microorganisms, comprising an insoluble substrate; and a carbohydrate receptor immobilized on the insoluble substrate, the carbohydrate receptor being capable of adsorbing microorganisms; and a labelled reagent useful for detecting the presence of microorganisms bound to the carbohydrate receptors and a method for detecting the presence of specified microorganisms in a sample, which comprises contacting a sample to be tested with carbohydrate receptors immobilized on an insoluble substrate; and determining the extent of binding of microorganisms in the sample to the carbohydrate receptors by use of a labelled reagent.

Abstract: Particle patterns formed on a conical bottom surface of wells formed in a microplate are photoelectrically detected to produce a two-dimensional image signal, and then the two-dimensional image signal is processed to judge or classify the particle patterns as agglutinated patterns, non-agglutinated patterns or uncertain patterns with the aid of the two-dimensional image processing. An image signal representing a particle pattern is first extracted, and then the true or typical agglutinated and non-agglutinated patterns are judged by means of at least two fast judgments. When the pattern is not definitely judged, the image signal of the relevant pattern is further subjected to at least two precise judgments. When the pattern is judged to be the agglutinated pattern by the precise judgments, the relevant pattern is finally judged to be the true agglutinated pattern.

Abstract: A method for drying mammalian cells, such as erythrocytes, lymphocytes, leukocytes and platelets onto a solid-phase support for use in solid-phase immunoassays through use of a drying solution and an article for use in immunoassays prepared by such method. The method comprises immobilizing a monolayer of cells onto the solid-phase support by non-covalent binding. This is accomplished by staining the solid-phase support with an organic dye having a net positive charge which permits non-covalent binding of the cells which carry a net negative charge to the solid-phase support. These cells are dried or fixed to the solid-phase support by addition of a drying solution which comprises an aqueous solution of a monosaccharide, disaccharide, trisaccharide or cyclitol and a salt. The preferred monosaccharide is D-(-)glucose and the preferred salt is sodium chloride. The preferred drying solution comprises a 1.0 M solution of dextrose and a 154 mM solution of sodium chloride.

Abstract: Specified classes and subclasses of leukocyte blood cells are identified by immunohematology procedures, based on utilization of antigenic determinants on the cell surface, their reactivity with antibodies which fluoresce under known circumstances, and identified by utilization of principles of flow cytometry or morphology.This invention particularly concerns improvements in the lysing and fixing method used prior to detection and identifying of the cells. In this method, a whole blood sample first is incubated with a reagent including antibodies to the cell subclass to be identified, the antibodies having directly or indirectly made fluorescently responsive to a particular light (e.g. argon ion laser). The red blood cells then are lysed with a reagent containing saponin. Next follows a leukocyte fixing treatment, preferably using a cross-linking dialdehyde composition, such as glyoxal or glutaraldehyde.

Abstract: Methods for diagnosing pre-hypertension, hypertension, congestive cardiomyopathy, renal failure, salt-sensitivity and adenomas and endocrine cell hyperplasias are disclosed. Also disclosed are methods for monitoring hypertension therapy, congestive cardiomyopathy therapy, renal failure therapy and adenoma and endocrine call hyperplasia therapy. These methods involve using an antibody having binding specificity to ouabain to immunologically measure the level of human ouabain in body fluid or tissue of a subject. Additionally, methods for treating a hypertensive subject by inducing passive or active immunity to human ouabain in the subject are disclosed, along with an antibody having binding specificity for ouabain.

Abstract: A new ligand binding assay is based upon measurements of relaxation rates of the solvent, which are obtained with a magnetic resonance (MR) spectrometer. It is termed a solvent mediated relaxation assay system (SMRAS). SMRAS is based on the observation that the enhancement of proton relaxation rates produced by a magnetic material can be modulated by the binding of various analytes to a magnetic material. Hence the relaxation rate of the solvent can be interpreted to give information on the concentration of an analyte.

Abstract: Method using tetra-aryl porphyrins for and, in particular, 5,10,15,20-tetrakis(4-carboxyphenyl)porphine as a fluorescent tracer for cancers of the lung, and as a radiotracer therefor as a complex with .sup.67 Cu. The latter complex also provides a source of beta radiation for selective destruction of lung malignancies as well as gamma radiation useful for image analysis of the situs thereof by single photon emission computed tomography, as an example, both in vivo. Copper-64 may be substituted for the .sup.67 Cu if only radiotracer characteristics are of interest. This lighter isotope of copper is a positron emitter, and positron emission tomography techniques cna be used to locate the malignant tissue mass.

Abstract: The present invention describes a class of dyes for use in staining cell samples and methods of making such dyes. A preferred class of dyes known as detergent dyes which possess the ability to stain cells in whole blood and are only slowly leached or lost from the stained cells over time are described. The present invention has application, for example, to blood typing for the determination of the presence of blood group antigens A, B, AB, O, and D (Rh.sub.o) and antibodies to such antigens.

Abstract: Microcapsule-reagents are prepared by previously reacting at least a part of an antigen or antibody attached to microcapsules encapsulating a labeling substance with an antibody or antigen which is specifically reactive therewith, and then the reagent thus prepared is reacted with a sample containing an antigen or antibody in the presence of a complement, whereby highly sensitive immunoassay of the antigen or antibody in the sample can be realized even when the antigen or antibody attached to the microcapsules has a lowered activity or has only low reactivity with the antibody or antigen in the sample.

Abstract: A solid phase immuno-assay system for assaying at least one analyte, in the form of a solid support having a plurality of receptors bound thereto. At least two of the receptors conjugate with the same analyte.A reaction container comprising a plurality of longitudinally arranged individual compartments, and a longitudinally extending single compartment.A card for assaying a plurality of samples for the same analyte, having a plurality of receptors for the analyte at different locations on the card.A method of performing an assay for the same analyte in more than one sample, by providing a receptor for the analyte at more than a single location on a solid substrate; exposing each of the receptors to different samples; and developing each of the receptor locations to indicate the presence of the analyte in each of the samples.

Abstract: A flow cytometry method for reproducibly detecting and counting a lymphocyte population of interest in a leukocyte suspension or whole blood sample in which the red cells are subsequently lysed. The suspension (or sample) is combined with a reagent comprising a primary antibody, either native, carrying an attached enzyme or biotin or other label, and a fixative reagent, in either order. Where the enzyme is not attached, an enzyme is coupled specifically to the primary antibody. The fixed suspension is reacted with a color-producing enzyme-cytochemical reagent. The suspension, now including stained and unstained fixed cells, is passed through a flow cytometer and the cells are characterized and counted on the basis of their light-scattering and light-absorbing properties.

Abstract: The present invention provides a one-step process for the detection of the presence of an allergy and for the specific detection of the allergen responsible for the allergy, in which the leukocytes of a sample to be investigated are incubated with an allergen or with another stimulation factor in an aqueous medium together with a chromogenic protease substrate and calcium ions, the liberated protease is reacted with the chromogen and the resulting chromophor is determined. The protease activity is measured kinetically after an incubation period by the increase of the chromophor concentration. The present invention also provides a reagent and a device for carrying out this process, as well as a process for the determination of antiallergic and anti-inflammatory substances. The device consists of a microtiter plate with a plurality of different reagents arranged in rows.

Abstract: There is disclosed, in an immunoassay employing an immunoassay reagent comprising liposomes comprised of at least any one of phospholipids and glycolipids, a marker material enclosed in said liposomes, and an antibody or part of the antibody, or an antigen, immobilized on said liposomes by a cross-linking method, the improvement wherein a material for preventing nonspecific lysis of the liposomes is made present together in a reaction mixture.

Abstract: An antibody matrix immunoassay device for determination of number and proportion of subpopulations of leukocytes is described. The device can be used to evaluate and monitor the immune status of an individual and to diagnose and monitor the progression of AIDS.

Abstract: In a novel, erythrocyte agglutination assay, the agglutination reagent comprises at least one erythrocyte binding molecule coupled to at least one specific analyte binding molecule wherein the erythrocyte binding molecule does not cause agglutination when incubated with erythrocytes in the absence of analyte (in the case of a direct assay) or analyte binding reagent (in the case of an indirect assay). Preferably, the erythrocytes are endogenous to the blood sample to be tested, that is, a whole blood sample is assayed. Mixtures of conjugates and conjugates of analyte analogues with erythrocyte binding molecules may also be used as agglutination reagents. The reagents and their use in direct or indirect assays is disclosed.

Abstract: Antigens specific to respiratory syncytial virus are produced on the surface of cells by:(1) culturing in vitro cells derived from a human or animal mucosa,(2) inoculating the cultured cells with respiratory syncytial virus, and(3) selecting virally infected cells from the culture.The resulting cells or the viral antigen(s) when partially or completely isolated from the cells have immunological and diagnostic uses in respect of infection by respiratory syncytial virus and may be used to isolate viral antibodies. A specific cell strain NM7 produced by this method from bovine nasal mucosal cells has respiratory syncytial virus antigens on its surface and its corresponding, uninfected cell strain NM5 can be infected similarly.

Abstract: A method of identifying and enumerating specific cell types in a heterogeneous population of cells by enhancing the specific staining of desired cells, comprising contacting a sample from the heterogeneous population of cells with a labeled primary antibody which recognizes and binds to a desired cell surface antigen and an unlabeled cross-linking agent which recognizes and binds to the primary antibody is disclosed.

Abstract: Assay methods and compositions are provided for determining an analyte in a sample suspected of containing the analyte. The composition comprises in a novel single liquid reagent at least one specific binding pair (sbp) member and its complementary member wherein at least one sbp member is reversibly confined in a material that temporarily renders the confined sbp member incapable of binding with its complementary sbp member. At least one of the sbp members is bound to a member of a signal producing system capable of producing a detectable signal in relation to the amount of analyte in the sample. The confinement is reversed, any remaining members of the signal producing system are added, and the signal produced in relation to the amount of analyte is measured. Examples of the confining material are lipid bilayers, cells and gels.

Abstract: A method of binding an antibody with protein A cells is provided which includes a sequence of incubation and dilution steps to produce a preselected amount and concentration of antibody with a preselected distribution of antibody among the protein A cells. In addition, a method is provided for preparing an antibody entrapped porous matrix and apparatus which includes a specific porous matrix with a preselected position of antibody bound bacterium cells therein along with means for drawing fluids through the porous medium and means for facilitating the deposition of fluids onto the surface of the porous matrix. The apparatus and method is useful for testing for the level of progesterone in animal body fluids, such as milk, plasma, serum, whole blood and saliva.

Abstract: A method for multi-parameter analysis of cells in a body fluid sample is described which makes use of a plurality of fluorescence measurements, comprising at least two nucleic acid dyes and at least one fluorescently labelled cell surface marker, and a plurality of light scattering measurements. A kit containing the nucleic acid dyes and cell surface marker also is described.

Abstract: The present disclosure described an assay for screening monoclonal antibodies for their potential as highly cytotoxic immunotoxins. The assay involves treating cells with dilutions of the test antibody followed by a Fab fragment of a secondary antibody coupled to an A chain toxin ("indirect assay"). The cytotoxicity of the indirect assay is compared to that of the direct assay where the monoclonal antibody is coupled to an A chain toxin. Indirect and direct assays were carried out using 14 antibodies and a panel of 8 human and mouse cell types. The two assays showed virtually 100% correlation. The indirect assay, therefore, predicts the potency of a given monoclonal antibody to make an effective immunotoxin and should be useful in screening monoclonal antibodies for use as immunotoxins.

Abstract: A method for detecting pathogen infection in a host is provided. The method comprises lysing phagocytes from the host to release soluble components of the pathogen which are detected subsequently using a specific binding assay.

Abstract: A substance-conjugated complement component C1q is provided. A substance such as signal emitting substances or cell function regulating substances is conjugated via a sulfur atom to at least one site of the component. The site is not involved in binding immunoglobulins. A marker-labelled complement component C1q is used for measuring a complement-binding antibody, an antigen, a neutralizing antibody or a substance produced internally of and at the surface of a cell or a microorganism by measuring the marker.

Abstract: Monoclonal antibodies reactive with oncogenic and activated ras p21 proteins containing aspartic acid or valine at position 13 and unreactive with normal ras p21 proteins containing glycine at position 13. The antibodies are secreted by hybridomas obtained by immunizing mice with synthetic peptides corresponding in amino acid sequence to positions 5-9 of normal ras p21 proteins, except having aspartic acid in place of glycine at position 13 and except for the addition of cysteine at the amino terminal end of the peptide. The antibodies and immunoreactive fragments are useful in the classification of malignant and premalignant lesions.

Abstract: This invention relates to a flotation immunoassay employing a novel buoyant matrix to which an antigen or antibody is coupled and which separates the bound and free products of the assay by floating to the surface of the reaction liquid. The novel flotation device which makes it possible to detect and to quantitate either antigen or antibody can also be used to fractionate cells and molecules.

Abstract: A method of measuring chain breakage in the DNA of a eucaryotic cell is disclosed. This method includes (a) contacting the cell with a stripping solution that lyses and solubilizes the cell without denaturing its DNA, thereby forming a nucleoid having a halo, (b) measuring the width of the halo, and (c) determining the number of chain breaks from the measured width. The halo includes at least one loop of undenatured DNA, and has a width related to the number of DNA chain breaks in the loop. The cell to be examined may be adhered to a support prior to its contact with the stripping solution. The number of DNA chain breaks can be determined by compairing the measured width of the nucleoid halo with a reference value.

Abstract: Biological cells which are fluorophore-labeled can be stored frozen for a long period of time. After thawing the cells can be used as targets in fluorophore-release cytotoxicity assays.

Abstract: A reagent for reticulocyte counting by flow cytometry which comprises two solutions, namely, a stock solution for staining in which a dye is dissolved in a nonaqueous solvent, and a buffer solution which satisfies the optimum staining conditions.By combining these two solutions immediately before measurement, a stable final staining solution for reticulocyte counting can always be obtained.

Abstract: A reagent for reticulocyte counting by flow cytometry characterized in that it contains a carbonate salt.If a blood sample is left exposed to ordinary air, non-specific staining of erythrocyte can be prevented with the addition of not less than about 1 mM of NaHCO.sub.3.

Abstract: A solution of an active protein substance and an inactive protein substance is reacted with a cross-linking agent, optionally in the presence of an inert carrier, under cross-linking conditions to produce articles comprising both active and inactive protein substances. The active protein substance comprises up to about 20 percent, e.g. from 1 to 20 percent by weight, based on the final weight of the total protein substance, whereas the cross-linking agent comprises from 0.5 to 8 percent by weight, based on the weight of the total treated mixture. The obtained articles are in the form of a solution or a suspension in aqueous medium, in the form of a film, in the form of a membrane, in the form of a fabric, in the form of a porous material, or in the form of a mass, such as granules, pills or tablets.

Abstract: An article for performing solid-phase immunological assays and method for performing assays utilizing the article of the invention. A solid-phase support suitable for immunological assays is stained with an organic dye that has a net-positive charge and a hydrophobic aromatic ring structure. An immunologically reactive component having a net-negative charge is immobilized by the dye to the support through non-covalent interactions. The immunologically reactive component is tightly bound by the organic dye. This is advantageous when performing immunological assays utilizing automated washing instruments. A sample of a biological fluid is added to the article having the immunologically reactive component bound thereto to detect the presence of antigens or antibodies in the biological fluid specific for the bound immunological component. The presence of the antigen-antibody complex may be determined by any known method.

Abstract: Methods and materials for preparing specific binding reagents with a multiplicity of relatively noninterfering label moieties are described. By spacing the labels at the surface of a specific reagent with bulking agent, increased sensitivity can be achieved without interference between individual labeling entities.

Abstract: A sac, in particular a vesicle, having a detectable metal, in particular a rear earth metal, encapsulated therein. The sac may be sensitized with a ligand and used in an assay. The vesicle is stored and/or used in an aqueous solution which includes the rare earth metal to increase the concentration of metal in the sac.

Abstract: A flow through test device and assay kit suitable for use by unskilled technicians is described. The flow through device has a porous support and an absorptive layer. The device is constructed for control of flow rates so that a tracer having a visible particulate label can be used. Optional flow control and porous spacer layers may be included.

Abstract: Methods and associated devices for detecting the presence of a particular motile organism within a sample are disclosed. The sample may be derived from dry milk, raw meat, poultry or a clinical specimen. A preferred method includes inoculating a selective enrichment medium containing a chemotactic attractant with the sample and contacting the selective enrichment medium with a nonselective motility medium containing a chemotatic attractant in a concentration less than the attractant concentration in the selective enrichment medium. Upon incubation, the motile organism metabolizes the chemotactic attractant, allowing the organism to move into the motility medium where it interacts with antibodies specific for the organism, thereby causing the formation of a persistent immobilization band. The methods are particularly useful in detecting Salmonella.

Abstract: Methods for delivering substances into, removing substances from, or reacting substances with a selected environment utilizing polymer gels or coatings characterized by a critical solution temperature (CST) are disclosed. The CST as well as the pore structure, pore size, pore distribution, and absorbing capacity of the gel may be selectively controlled. The substances may be physically or chemically immobilized within the polymer gels. In addition, a method for altering the surface wettability of CST polymers is also disclosed.

Abstract: A method of determining the presence and quantity of an analyte of interest in a particulate-containing sample is disclosed, as is a construct for use in the method. The method is particularly useful for determining an analyte in whole blood and in fermentation suspensions. The construct is comprised of a first moiety, which is a particulate-binding moiety and a second moiety, which binds the analyte of interest.

Abstract: A test card incorporates a colorimetric indicator test to determine the presence of a minute amount of a specific substance in a liquid medium. The test card includes top and bottom sheets adhesively secured to an intermediate frame member which, together with the top and bottom sheets, defines a filter chamber having a flat filter therein which incorporates a test portion. The test portion of the filter has a binding substrate to which antibodies of the specific substance have been bound. This binding substrate is in communication with a test port. In the method of the invention a substance to be tested is administered through the test port and contains an unknown amount of antigen. After the sample is administered an aqueous solution of enzyme labelled antigen is administered and, subsequently, a liquid substrate is added to cause a color change in the filter below the test portion inversely proportional to the amount of antigen contained in the sample administered at the test port.

Abstract: Proteinaceous, antigenic factor derived from a group C Streptococcus which is receptor for the Fc region of IgG, a method for its preparation and immunoassay and antigen detection methods employing the receptor.

Abstract: This invention provides an immunologic method for measuring plant pollen germination and pollen tube growth. This method is based on the finding that an antibody having specificity to .beta.-L-arabinofuranosyl residues binds to the surface of in vitro grown pollen tubes. Antibody binding can be measured qualitatively or quantitatively using a calibration curve. The immunoassay is useful for screening plants for response to substances that affect plant germination or pollen tube growth.

Abstract: In an erythrocyte agglutination assay, the agglutination reagent comprises at least one erythrocyte binding molecule coupled to at least one specific analyte binding molecule wherein the erythrocyte binding molecule does not cause agglutination when incubated with erythrocytes in the absence of analyte. Preferably, the erythrocytes are endogenous to the blood sample to be tested, that is, a whole blood sample is assayed. Mixtures of conjugates and conjugates of analyte analogues with erythrocyte binding molecules may also be used as agglutination reagents. The reagents and their use in direct or indirect assays is disclosed.

Abstract: Assay methods are provided for determining an analyte in a sample suspected of containing the analyte. The method is carried out using a composition that includes a conjugate of a first sbp member with a particle. A luminescer is reversibly associated with a nonaqueous phase of the particle. Where the first sbp member is not complementary to the analyte, a second sbp member that is capable of binding to the first sbp member is employed. Unbound conjugate is separated from conjugate that is bound to the analyte or to the second sbp member. A reagent for enhancing the detectability of the luminescer is added and the light emission of the luminescer acted on by the reagent is measured.

Abstract: The present disclosure is directed to methods for the preparation of immunoadsorption matrices having IgG molecules adsorbed thereto in a preferred configuration, i.e., adsorbed to the matrix by their (Fc) rather than F(ab) portions. IgG molecules, are selected such that the F(ab) portion of the IgG fraction adsorbed has a more acidic or basic net isoelectric point or pI range than the F(c) end of the molecule, depending on the characteristics of the adsorption surface. For negatively charged surfaces, IgG molecules having relatively alkaline F(c) portions are selected. For positively charged surfaces, IgG with relatively acidic F(c) portions are selected. Additional selection criteria include pI fractionation to provide fractions having well defined pI characteristics as defined by "non-overlap" or "pI range" of F(c) and F(ab) portions pI's. Methods disclosed are particularly well suited to the preparation of colloidal gold immunostains.

Abstract: Apparatus and method for providing an optical detection of a binding reaction between a ligand and an antiligand, including, a pattern formed by a spatial array of microscopic dimensions of antiligand material, ligand material interacting with the antiligand material to produce a binding reaction between the ligand and the antiligand in the pattern, a source of optical radiation including energy at at least one wavelength directed to the pattern at a particular incidence angle to produce scattering of the energy from the pattern in accordance with the binding reaction and with a strong scattering intensity at one or more Bragg scattering angles, and at least one optical detector located relative to the pattern and aligned with a Bragg scattering angle to detect the strong scattering intensity at the Bragg scattering angle to produce a signal representative of the binding reaction.

Abstract: A test kit useful for the determination of Streptococcus A antigen comprises: (i) an immunoreactive reagent comprising either Streptococcus A antigen or antibodies attached to water-insoluble particles, (ii) a substrate having thereon a dried, binder-free coating of a first extraction reagent, (iii) an aqueous solution of a second extraction reagent, and (iv) a neutralizing solution. Both extraction reagents combine to provide nitrous acid. In addition, an extraction device includes a water-insoluble container having affixed therein the first extraction reagent and an applicator means for collecting and depositing a biological specimen within the container. The extraction device and test kit are useful to the determination of Streptococcus A antigen in a biological specimen.