Abstract: The present invention discloses a method of monitoring chemotaxis or chemoinvasion comprising: providing a device for monitoring chemotaxis. The device includes a support member; a top member mounted to the support member by being placed in substantially fluid-tight, conformal contact with the support member. The support member and the top member are configured such that they together define a discrete assay chamber. The discrete assay chamber includes a first well region including at least one first well, the at least one first well being configured to receive a soluble test substance therein; a second well region including at least one second well, the at least one second well being configured to receive a sample comprising cells therein; and a channel region including at least one channel connecting the first well region and the second well region. Preferably, at least one second well is horizontally offset with respect to the first well region in a test orientation of the device.

Abstract: The T cell activation marker, granulysin, is demonstrated to be an effective antimicrobial agent. It is used in vitro and in vivo to reduce the population of viable cells in a microbial population. Of particular interest is the use of the active fragment of human granulysin, or modified forms thereof, to treat bacterial infections.

Abstract: An automated system and method for loading individual cells into individual discrete locations. The system includes a cell carrier grid, a cell carrier grid holder, a vacuum source, a liquid reservoir and a loading device facilitating communication between the above components. Application of vacuum via a port causes cells to move into individual discrete locations. The method includes the steps of placing the grid holder into a loading device, automatically filling a space in the grid holder with a liquid, automatically adding to an upper surface of the grid and automatically applying a force to the cells in so that individual cells enter at least some of the individual discrete locations. Further disclosed is an automated system for collection of data from cells further including an electro-optical scanner capable of illuminating the cells and collecting at least a portion of photons therefrom and a computerized control mechanism further controlling same.

Abstract: A diagnostic microbiological testing system and method for microorganism identification (ID) and antimicrobial susceptibility determinations (AST). The system includes multiple-well test panels capable of performing ID and AST testing on the same test panel. Each test panel is inoculated with reagents, broth-suspended organisms, and placed into the instrument system. The instrument system includes a rotating carousel for incubation and indexing, multiple light sources each emitting different wavelength light, precision calorimetric and fluorometric detection, barcode test panel tracking and a control processor for making determinations based on measured test data. One light source includes a plurality of LEDs arranged in a linear array. Each of the LEDs' junction currents are controllable to produce a predetermined illumination profile.

Abstract: The present invention concerns methods of testing water for microbe contamination. The methods of the invention comprise supplementing existing methods with assays using specific reagents such as monoclonal antibodies. The invention also concerns a device for use in the methods of the invention.

Abstract: High-throughput screening method and apparatus are described. The method includes placing cells on a substrate defining a plurality of discrete microwells, at a well density of greater than about 100/cm2, with the number of cells in each well being less than about 1000, and where the cells in each well have been exposed to a selected agent. The change in conductance in each well is determined by applying a low-voltage, AC signal across a pair of electrodes paced in that well, and synchronously measuring the conductance across the electrodes, to monitor the level of growth or metabolic activity of cells contained in each well. Also disclosed is an apparatus for carrying out the screening method.

Abstract: The invention provides exemplary devices and methods to facilitate the performance of assays. In one embodiment, one such device comprises a holding member having a top surface, a bottom surface, and a plurality of holding locations that are adapted to hold at least one article, such as a solid support and/or a cell. When within the holding locations, the articles are preferably disposed below the top surface. A membrane is positioned above the top surface of the holding member, and a pressure system is provided to apply positive pressure to the membrane to force the membrane against the top surface of the holding member. In this way, a seal may be provided between the holding locations.

Abstract: A method and an apparatus for determining the concentration at which a growth-altering agent has an appreciable effect on the growth of a target microorganism are provided.

Abstract: The invention relates to a method and apparatus for detecting specific target cells in a simple and time-saving way, using paramagnetic particles, antibodies recognizing the Fc portions of target-cell associating antibodies and target-cell associating antibodies directed to specific antigen determinants in the target-cell membranes. Incubation of the cell suspension with detergent and/or second antibodies or antibody fragments, prelabeled or not with fluorescent agents, metallocolloids, radioisotopes, biotin complexes or certain enzymes allowing visualization, dramatically increase the specificity of the method. The method and apparatus described provides a solid support and permanent record which is easily viewed by microscopy, permits viewing and quantification of the whole specimen rather than small fractions thereof and allows the use of large specimen volumes to be analysed, the device may also be scanned automatically by conventional densitometric technology.

Abstract: Substrates are patterned with antibodies attached thereto at discrete locations from which absorption resistant coating is removed by selectively controlled mechanical scribing contact to avoid chemical removal so as to decrease fabrication costs and increase fabrication speed.

Abstract: A microtitration unit being at least one container of transparent material for accommodating biological test reaction mixtures. The container is subdivided into at least two chambers suitable for accommodating different test reaction mixtures by at least one partition which extends from the inside of the bottom of the container and which is of lower height than the internal height of the container.

Abstract: Systems including (1) a micromatrix and perfusion assembly suitable for seeding and attachment of cells within the matrix and for morphogenesis of seeded cells into complex, hierarchical tissue or organ structures, wherein the matrix includes channels or vessels through which culture medium, oxygen, or other nutrient or body fluids can be perfused while controlling gradients of nutrients and exogenous metabolites throughout the perfusion path independently of perfusion rate, and (2) sensor means for detecting changes in either cells within the matrix or in materials exposed to the cells, have been developed. Methods for making the micromatrices include micromachining, micromolding, embossing, laser drilling, and electro deposition machining. Cells can be of one or more types, either differentiated or undifferentiated.

Abstract: A method and apparatus for evaluating the effects of toxic substances in the environment on aquatic organisms is disclosed. The apparatus allows for exposure of test organisms, placed in test chambers made of partially submerged sieves, to water that is pumped from an urban creek at preset time intervals. The invention also allows for maintenance of cool temperatures in the exposure chambers, both for the treatment (i.e., creek water) and the experimental control.

Abstract: Disclosed is a novel biological sample preparation (BSP) device and a method of using such a device that enables inexpensive and flexible high throughput sample preparation and visualization for microscopy, including high resolution, single or multi-label, 2-dimensional (2D) or 3-dimensional (3D) fluorescence microscopic observation of biological samples, such as cultured cells. Also included in the invention are a high throughput chemical sample preparation device, and a method for to clone and screen cells using a BSP device.

Abstract: A microbiological test panel assembly used in microorganism identification (ID) and antimicrobial susceptibility determinations (AST) testing is provided. The microbiological test panel assembly includes a plurality of test wells segregated into two sections. The test wells of each section are adapted to receive reagents capable of causing reactions used in performing ID and AST testing. The reagents enter the respective sections through fill ports and flow down a passageway of the test panel assembly in a serpentine manner filling all the test wells.

Abstract: A computer interfaced device for rapidly quantifying levels of toxicity to small aquatic organisms in aqueous environments. The device has a plurality of test chambers containing replicates of a plurality of concentrations of a substance or sample being tested for toxicity. Each chamber also contains a predetermined number of motile aquatic organisms. Light originating on one side of each test chamber is received by a photo-detector on the opposite side. The device's sensitivity is adjusted so that each interruption of a light beam caused by movement of a test organism in each test chamber is recorded for that chamber. Using this device and a toxicity testing protocol both lethality and changes in the frequency of test organism's movement in each concentration of substance or sample is monitored. Toxicity is by convention quantified as the concentration of a substance or sample which causes a predetermined level of lethal or sublethal response usually after a plurality of days.

Abstract: A patterned multiple antibody substrate for use in biosensors or immunosensors is produced by (1) coating an antibody-adsorbent substrate with a material that resists antibody adsorption, (2) using ion beam sputtering, laser ablation, or mechanical scribing to remove the coating at specific sites on the substrate, and then (3) adsorbing specific antibodies at the sites. The substrate is capable of detecting multiple chemical species simultaneously.

Abstract: An article adapted for holding a liquid sample for quantification of biological material in the liquid sample. The article includes a bag having an upper surface sheet and a lower surface sheet enclosing a volume therebetween. The bag has an upper opening through which the liquid sample can be poured into the volume in the bag. The bag also has a plurality of partitions configured to separate one or more portions of adequate sample in the bag. Also provided is a passage through which a liquid sample can be distributed throughout the volume in the bag. The bag is made of material which can be caused to form discreet non-permeable compartments for holding separate aliquots of the liquid sample.

Abstract: A biosensor for detecting the toxicity of a sample includes a solid-state area image pickup element, a culture container positioned on an upper surface of a light-receiving portion of the element, at least a bottom surface of the culture container being formed of transparent material, a cell cultured in the culture container, and culture medium for growing the cell.

Abstract: Procedure and relative equipment for determining the presence of toxic substances in water destined for drinking and/or in water effluents, based on the inhibition of the production of biogas (methane) on the part of anaerobic bacterial aggregates.

Abstract: A test solution including bacteria is added to a liquid sample in which antibacterial drug exists and to a liquid sample in which no antibacterial drug exists, and existence or non-existence of drug sensitivity is judged based upon whether or not the dissolved oxygen concentration in both liquid samples are essentially different from one another.

Abstract: An article adapted for holding a liquid sample for quantification of biological material in the liquid sample. The article includes a bag having an upper surface sheet and a lower surface sheet enclosing a volume therebetween. The bag has an upper opening through which the liquid sample can be poured into the volume in the bag. The bag also has a plurality of partitions configured to separate one or more portions of adequate sample in the bag. Also provided is a passage through which a liquid sample can be distributed throughout the volume in the bag. The bag is made of material which can be caused to form discreet non-permeable compartments for holding separate aliquots of the liquid sample.

Abstract: The present invention relates to an in vitro system for physiological and metabolic evaluation of substances for use in living beings. The system includes one or more cell culture chambers, each containing cells in a culture medium and a gas-liquid exchange device for contacting the culture medium with oxygen-containing gas so that the culture medium absorbs that gas and desorbs carbon dioxide-containing gas. The conduit system conducts culture medium between the gas-liquid exchange device and the cell culture chambers. A circulation mechanism is used to circulate culture medium through the conduit system, the cell culture chambers, and the gas-liquid exchange device. In use, the substance to be evaluated is added to the culture medium of the system and circulated through the system. The cells in each of the cell culture chambers are then evaluated for effects resulting from the presence of the substance.

Abstract: A moist unitary non-laminar chromogen agar color reactive test sheet for use in a simplified enzyme-linked immunosorbent assay (ELISA) for the diagnosis of viral, fungal, bacterial and parasitic diseases. The diagnostic test is designed for use in non-laboratory settings where the usual equipment and supplies, including running water, may not be available. The test sheet comprises chromogen agar paper (CAP) impregnated with an enzyme substrate, and electron acceptor, if needed, in agar. It is applied over a conjugate in the form of a species specific enzyme-linked anti-immunoglobulin bound to an antibody-antigen coating of a previously prepared test plate. After incubation to cause a color reaction, the results are read and interpreted in comparison with known standards.

Abstract: A method and apparatus for assaying the sensitivity of biopsied tumor cells to chemotherapeutic agents using a predetermined amount of chemotherapeutic agent in an easily deliverable form is disclosed and claimed.

Abstract: A microculture tray comprising a plurality of pairs of wells, each pair of wells connected by a trough which extends downwardly approximately 2/3 of the depth of the well so that the wells in the pair are connected so as to allow free passage of supernatant but not of cells between the wells.The tray can be used for rapid testing of animal cells or non-motile microorganisms for the release of soluble mediators.

Abstract: A method and apparatus for a high time resolution study of a reaction pattern of a cell or cell aggregates by light transmission microscopy during perfusion by different media. The apparatus comprises a light transparent chamber defining a first space for receiving a medium and a second space for receiving a cell or cell aggregates. The first space and second space are separated by a transparent slide and communicate with one another through a slit shaped opening. The slit shaped opening allows a first medium in the first space, for instance a control medium, to be drawn through the cell space by an external pump. The control medium in the first space may be replaced by a different medium, for instance a cytotoxic drug, by pumping the control medium from the first space and replacing it with the test medium. Pumping the control medium out of the first space however does not remove the control medium from the second space.

Abstract: Normally transcriptionally silent genes in a cell line or microorganism may be activated for expression by inserting a DNA regulatory element which is capable of promoting the expression of a normally expressed gene product in that cell or which is promiscuous, the regulatory element being inserted so as to be operatively linked with the normally silent gene in question. The insertion is accomplished by means of homologous recombination by creating a DNA construct including a segment having a DNA segment of the normally silent gene (targeting DNA) and the DNA regulatory element to induce gene transcription. The technique is also used to modify the expression characteristics of any endogenous gene of a given cell line or microorganism.

Abstract: A method and apparatus for assaying the sensitivity of biopsied tumor cells to chemotherapeutic agents using a predetermined amount of chemotherapeutic agent in an easily deliverable form is disclosed and claimed.

Abstract: A method for the rapid and reliable in vitro determination of the anticoccidial activity of chemotherapeutic agents. The invention involves incubating free-living sporozoites in a nutrient medium in the presence of the anticoccidial agent being assessed and 2,3,5-triphenyltetrazolium chloride. Living sporozoites reduce the above compound to the water-insoluble vivid-red formazan pigment but will not stain with trypan blue; dead sporozoites do not form this red pigment but will stain with trypan blue.

Abstract: An apparatus and method for monitoring cell-substrate impedance comprises upper and lower substrates each containing strips of metal foil which cross to form well areas. Holes in the upper substrate expose an insulation layer between the upper and lower substrates. Smaller holes in the insulation layer expose the conductive material of the lower substrate. Cylindrical well walls are positioned around each of the holes in the upper substrate and define a cell medium containing well which is exposed to an annular area of conductive foil on the upper substrate, and a small area of conducive foil on the lower substrate. The areas of foil form large and small electrodes which are positioned in an array of wells which can be used to monitor the impedance of various cell cultures under various conditions. A switching device applies currents to the well electrodes and a monitoring device measures the impedance between the electrodes in each well. In this way, cell spreading and motility can be measured.

Abstract: An improved simplified enzyme-linked immunosorbent assay (ELISA) for the diagnosis of viral, fungal, bacterial and parasitic diseases. The diagnostic test is designed for use in non-laboratory settings where the usual equipment and supplies, including running water, may not be available. Collected blood samples are applied to a previously prepared test plate having an antigen coating covered by a layer of agar. After incubation to cause antibodies from the samples to bind to the antigen coating, the agar layer is stripped off and a conjugate is applied in the form of a species specific enzyme-linked anti-immunoglobulin. After incubation to cause this conjugate to bind to the antibody-antigen coating, a previously prepared chromogen agar paper (CAP) impregnated with an enzyme substrate and acceptor, if needed, in agar is applied over the conjugate. After incubation to cause a color reaction, the results are read and interpreted in comparison with known standards.

Abstract: The present invention relates to a method for determining the antibiotic sensitivity of pathogenic microorganisms present in milk obtained from mammals afflicted with mastitis, an apparatus for use with the same and an antibiotic sensitivity test kit containing the same.

Abstract: A multiplate subculture solid media device is provided with a connection for internal flow communication with a liquid medium culture bottle. The device includes a large transparent screw cap on one face thereof to gain easy access to the plurality of media plates for direct examination and sample taking through the growth period. However, the cap is transparent and of a size allowing continuous examination during the growth period without any opening. As a further feature of the invention, the screw cap may include an integral magnifying lens for enhancing the continuous examination of culture growth.

Abstract: A test set comprising a purified culture of Streptococcus thermophilus T101 concentrate and a water-based protective agent in dilution ratio of about 4 to 5.times.10.sup.-2. The test set may also advantageously comprise an indicator. The test method includes the steps of incubating the test set and evaluating the color. The present invention also comprises a purified culture of Streptococcus thermophilus T101 strain.

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: A virus-free assay to identify agents that interfere with the life cycle of HIV which comprises: (1) expressing in mammalian cells in the presence or absence of one or more of said agents, HIV gp120, the gene product of HIV tat, the gene product of HIV art/trs, all under the control of an HIV LTR promoter, and a control protein, not under the control of the HIV LTR promoter, such that infectious HIV particles are not produced; (2) quantitating the amount of the gp120 produced relative to the amount of control protein produced; and (3) comparing the relative amount of the gp120 produced in the presence of said one or more agents to the relative amount of the gp120 produced in the absence of said one or more agents.

Abstract: An anti-microbial sensitivity test and testing stratum is disclosed for evaluating the sensitivity of different anti-microbial agents relative to identifiable microbial contaminants. The test stratum comprises a polymer such as poly(2-hydroxyethylmethacrylate) referred to as PHEMA, an organic solvent, such as polyethylene glycol (PEG), a hydrogen binding plasticizer, such as dimethylsulfoxide (DMSO), an an anti-microbial agent such as silver sulfadiazine, mafenide, nystatin, nitrofurazone, silver nitrate, bacitracin, gentamicin, amphotericin B, cesium nitrate, or other anti-microbial agents. The test includes positioning testing stratum of the type described, each having a different anti-microbial agent therein, on a sterile plate or on an agar substrate and spaced in fixed relationship to one another. The testing stratum are self adhering and remain fixed on a sterile plate, or on an applicator, even when the sterile plate, is inverted.

Abstract: A substantially rigid, multi-well, self-pipetting device for use in chemical, immunochemical and microorganism analysis is provided. It includes a frame and a plurality of test wells disposed within the frame. A filing manifold communicates with each of the test wells through a filling passageway. A docking port on the periphery of the device receives a reservoir holding a fluid specimen. The docking port communicates with the filling manifold through a filling port and a filling channel. A vent control system also communicates with the test wells and is comprised of one or more vent manifolds, and/or valves to a vent manifold, and/or one or more vent holes to the ambient atmosphere. Any vent holes present are temporarily closed.

Abstract: A method and apparatus for photographically recording and determining fluorometric and nephelometric analysis are described.

Abstract: The presence of fermentation inhibiting activity in a sample is determined by contacting the sample with bacterial strains and detecting the inhibition of the activity of the strains. A carrier (1) carrying microtitration cups arranged in four rows of cups (8) is provided. The cups of the same row contain a lyophilisate comprising a given bacterial strain. Each sample to be tested, effected in a tube (3), is diluted in a tube (4) containing a culture medium and bromocresol purple and then poured into the cups of a given column by means of an automatic pipette and cones (5). After incubation, a drop of janus green reagent in tube (6) is introduced in each cup and, after a new incubation, the number of cups which have produced a change in color indicates the degree of contamination of the mixture. The process may be applied in the determination of bacteriophages present in cheese-making vats.

Abstract: An improved microbiological test tray is described which are provided with fasteners so that two or more of the trays may be joined together for ease in inoculation, incubation, lyophilization and reading. In another aspect of the invention an improved tray structure is described which provides openings for the flow of vapor during the lyophilization step so that a greater number of trays may be lyophilized at the same time and also that lyophilization may be carried out in a shorter period of time due to the improved vapor flow. In another aspect of the invention an improved test card is described in which the API 20 test strip is reshaped into a card of two rows of ten cupules conforming to the same pattern as other frequently used test trays. A holder tray is provided for the card and the holder is provided with connecting fasteners so that it and the test card may be joined into one or more additional test trays for ease in inoculation, incubation and reading.

Abstract: A process and agent are provided for testing the sensitivity of bacteria towards antibiotics with a primary action in murein biosynthesis. The process is characterized in that the cytoplasmic enzyme activities released from the bacteria in the presence of the antibiotic and an enzyme substrate are determined directly.

Abstract: A microdroplet test apparatus for use in HLA typing. The apparatus is an improvement upon the well known Terasaki tray which is in use for determining HLA antigens by measuring lymphocyte cytotoxicity. The apparatus includes a tray having a plurality of microtest wells wherein the sides of the microtest wells are designed to promote localization of lymphocytes or other cells being tested at the microtest well bottom. The apparatus further includes a cover having either rods or tubes which extend down into the test solution present in the microtest well to thereby control the amount of test solution which is present in the cylindrical optical view path through which the test solution is viewed by microscope for test result measurements.

Abstract: For microbiological and other laboratory work in which an active substance is contacted with a substrate there is provided a container for the substrate and a divided carrying body with respective separated sections in sealed contact within the container such that the sections form separate regions with isolated substrate portions in each of the regions. The active substance is supported by the carrying body and is immersed in the respective isolated substrate regions so that it can be diffused into the substrate portions.

Abstract: Disclosed is a method of screening or identifying a microorganism by providing two microorganism nutrient medium layers in communication with and separated by a sterilizable membrane filter, implanting at least one of the layers with a microorganism, and allowing an effective substance to permeate through the filter to enable microorganism identification.

Abstract: A convenient, self-contained, disposable device suitable for the rapid identification of microorganisms, having capability for simultaneously carrying out a multiplicity of analytical determinations. The device comprises a disc-shaped arrangement defining a multiplicity of satellite testing chambers each of which is connected to a centrally defined chamber. The central chamber is provided with a movable sleeve or piston arrangement. Liquid sample is received in the central chamber and the sleeve is manipulated to provide samples uniformly and simultaneously to each of the satellite testing chambers without resorting to external means, such as positive or a negative pressurization or centrifugal force. Also disclosed are unique disc-shaped optically transparent support films which are coated on one or both sides with analytical substrates or reagents which are predispensed to the satellite testing chambers.

Abstract: A biologically active substance is incorporated in a unitary body with a magnetically responsive material for carrying out diffusion testing. These may be, microbiological, immunological, serological and other biochemical examinations. The body is applied against a substrate or medium by application of an external magnetic field and a reaction region is produced at the site of the body and is measured by means of a reader. In order to insure deposit of the body on the substrate a predetermined location and corresponding reading of the reaction region at such location, the support for the substrate and the dispenser and rear are provided with suitable releasable coupling and orienting devices such that the dispenser and reader can be respectively engaged and oriented on the support in predetermined secured positions.

Abstract: An apparatus is described with which a plurality of microbiological tests made in wells located in a pattern of rows on a tray can be conveniently analyzed and evaluated. A housing is formed with a tray analysis section and over which an arm is mounted to move from row to row. A plurality of manually activated signal generators are mounted on the arm, with the generators being aligned with a well in a row to enable the operator, upon identification of a particular well in a row as representative of analysis, to actuate the signal generator in registration therewith. The signals are representative of a score value and are recorded on score forms located in a test scoring section over which the arm moves with recording elements, also mounted on the arm. An elongated illuminator for a row of wells is described and which is conveniently adjusted to view selected test wells against different background lighting.

Abstract: A medium for determining Escherichia coli sensitivity to a pre-selected antimicrobial agent. The medium contains nutrient sources, inhibitors to inhibit growth of gram-positive and of other gram-negative micro-organisms, and an indicator which will show metabolic activity or lack thereof by E. coli in the medium, and a specific antimicrobial agent whose effectiveness against E. coli is being tested. If a given agent is effective against E. coli when E. coli is added to the medium containing that agent, the medium will remain clear. If the specific agent is ineffective against E. coli the color of the medium containing E. coli and that agent will change to blue as the E. coli metabolizes in the medium and produces acid.