Abstract: Complex multidimensional datasets generated by digital imaging spectroscopy can be organized and analyzed by applying software and computer-based methods comprising sorting algorithms. Combinations of these algorithms to images and graphical data, allow pixels or features to be rapidly and efficiently classified into meaningful groups according to defined criteria. Multiple rounds of pixel or feature selection may be performed based on independent sorting criteria. In one embodiment sorting by spectral criteria (e.g., intensity at a given wavelength) is combined with sorting by temporal criteria (e.g., absorbance at a given time) to identify microcolonies of recombinant organisms harboring mutated genes encoding enzymes having desirable kinetic attributes and substrate specificity. Restriction of the set of pixels analyzed in a subsequent sort based on criteria applied in an earlier sort (“sort and lock” analyses) minimize computational and storage resources.

Abstract: Imaging hardware, software, calibrants, and methods are provided to visualize and quantitate the amount of Fluorescence Resonance Energy Transfer (FRET) occurring between donor and acceptor molecules in epifluorescence microscopy. The MicroFRET system compensates for overlap among donor, acceptor, and FRET spectra using well characterized fluorescent beads as standards in conjunction with radiometrically calibrated image processing techniques. The MicroFRET system also provides precisely machined epifluorescence cubes to maintain proper image registration as the sample is illuminated at the donor and acceptor excitation wavelengths. Algorithms are described that pseudocolor the image to display pixels exhibiting radiometrically-corrected fluorescence emission from the donor (blue), the acceptor (green) and FRET (red).

Abstract: The present invention provides mutant genes encoding variant galactose oxidase enzymes, termed vGOs; constructs and recombinant host cells incorporating the genes; the vGO polypeptides encoded by the genes; antibodies to the polypeptides; and methods of making and using all of the foregoing.

Abstract: Improvements in calorimetric assays, surfaces for arraying microcolonies and instrument hardware for screening mutagenized enzymes and proteins in a solid phase format are presented. These improvements permit new enzyme activities to be screened. New filter membrane materials and formats for arraying the microcolonies provide higher throughput, better solvent resistance and ease of handling. Modifications to the instrument heating and illumination systems provide improved temperature control and a more compact, folded light path.

Abstract: Imaging hardware, software, calibrants, and methods are provided to visualize and quantitate the amount of Fluorescence Resonance Energy Transfer (FRET) occurring between donor and acceptor molecules in epifluorescence microscopy. The MicroFRET system compensates for overlap among donor, acceptor, and FRET spectra using well characterized fluorescent beads as standards in conjunction with radiometrically calibrated image processing techniques. The MicroFRET system also provides precisely machined epifluorescence cubes to maintain proper image registration as the sample is illuminated at the donor and acceptor excitation wavelengths. Algorithms are described that pseudocolor the image to display pixels exhibiting radiometrically-corrected fluorescence emission from the donor (blue), the acceptor (green) and FRET (red).

Abstract: Imaging hardware, software, calibrants, and methods are provided to visualize and quantitate the amount of Fluorescence Resonance Energy Transfer (FRET) occurring between donor and acceptor molecules in epifluorescence microscopy. The MicroFRET system compensates for overlap among donor, acceptor, and FRET spectra using well characterized fluorescent beads as standards in conjunction with radiometrically calibrated image processing techniques. The MicroFRET system also provides precisely machined epifluorescence cubes to maintain proper image registration as the sample is illuminated at the donor and acceptor excitation wavelengths. Algorithms are described that pseudocolor the image to display pixels exhibiting radiometrically-corrected fluorescence emission from the donor (blue), the acceptor (green) and FRET (red).

Abstract: The invention relates generally to the area of drug development and more specifically to screening novel compounds for antimicrobial activity. Methods are described for generating mutagenized peptide libraries expressed in colonies of cells. A fluorescent live/dead cell assay combined with a digital imaging spectrophotometer provides a high-throughput solid-phase screening method for colonies or arrayed synthetic libraries. The assay enables screening of antimicrobial peptide activity in at least 105-106 colonies per experiment. These methods generate and identify antimicrobial compounds.

Abstract: The present invention provides mutant genes encoding variant galactose oxidase enzymes, termed vGOs; constructs and recombinant host cells incorporating the genes; the vGO polypeptides encoded by the genes; antibodies to the polypeptides; and methods of making and using all of the foregoing.

Abstract: Complex multidimensional datasets generated by digital imaging spectroscopy can be organized and analyzed by applying software and computer-based methods comprising sorting algorithms. Combinations of these algorithms to images and graphical data, allow pixels or features to be rapidly and efficiently classified into meaningful groups according to defined criteria. Multiple rounds of pixel or feature selection may be performed based on independent sorting criteria. In one embodiment sorting by spectral criteria (e.g., intensity at a given wavelength) is combined with sorting by temporal criteria (e.g., absorbance at a given time) to identify microcolonies of recombinant organisms harboring mutated genes encoding enzymes having desirable kinetic attributes and substrate specificity. Restriction of the set of pixels analyzed in a subsequent sort based on criteria applied in an earlier sort (“sort and lock” analyses) minimize computational and storage resources.

Abstract: A MicroColonyImager instrument and solid phase methods to screen cells expressing mutagenized enzymes for enhanced activity. The MicroColonyImager instrument and methods permit high throughput screening of enzyme libraries by time course analyses of single-pixels, using either absorption, fluorescence or FRET. This high throughput assay can detect small differences in enzyme rates within microcolonies grown at a nearly confluent density on an assay disk. Each microcolony is analyzed simultaneously at single-pixel resolution, requiring less than 100 ml substrate/measurement. By simultaneously assaying different substrates tagged with spectrally distinct chromogenic or fluorogenic reporters, the substrate specificity of an enzyme can be changed.

Abstract: A connector strip is disclosed for assembling a building wall of glass blocks. The strip includes a longitudinal center section having a pair of oppositely disposed and longitudinally extending grooves dimensioned to receive a centrally located ridge extending peripherally around the outer edge surfaces of a molded building block to which the strip is to be mounted. U-shaped channels extend along the center groove for receiving wiring such as fiber optics for illuminating the glass block wall. Peripheral members and secondary walls serve to strengthen the connector strip, engage marginal edge surfaces of glass blocks and facilitate cement bonding between the connector strip and the adjacent blocks. A fastener member also is provided for coupling together adjacently positioned connector strips to form a rigid framework for assembling successive rows of blocks into a wall assembly.

Abstract: A connector strip is disclosed for assembling a building wall of glass blocks. The strip includes a longitudinal center section having a pair of oppositely disposed and longitudinally extending grooves dimensioned to receive a centrally located ridge extending peripherally around the outer edge surfaces of a molded building block to which the strip is to be mounted. A flange projects laterally outwardly from each longitudinal edge of the center section. The flanges have outwardly profecting walls and a plurality of laterally extending ribs which project outwardly from the walls. The flanges, walls and ribs define a plurality of open compartments on opposite sides of the flanges. Resiliently compressible material is mounted to the center section which engages with the outer edge surface of the building block to which the strip is to be mounted.

Abstract: An improved pipe railing system is disclosed which employs a pair of basic structural parts which may be joined by means of a unique, integral fastening mechanism to form uniform horizontal pipe railings and vertical pipe posts which may be mechanically joined without the need for welding or industrial adhesives. The improved pipe railing structure is a pair of half-cylindrical hollow body members which each have first and second terminal edges with longitudinal fasteners for joining the two halves. A unique fastener anchorage member is disposed along the inner surface of each body member, having a longitudinal recess therein, for receiving and anchoring a fastener in the axial direction. In this manner, the lower body member of a top horizontal railing may be fastened to a vertical post made up of two such body members by means of simple screw fasteners, followed by mechanically snapping an upper cylindrical body member to the lower cylindrical body member thereby completing the top horizontal railing.