Abstract: An RNA tagging system for visualization of single mRNA molecules based on a MSB-MCP system, as well as methods of use.

Abstract: An RNA tagging system for visualization of single mRNA molecules based on a MSB-MCP system, as well as methods of use.

Abstract: Described herein is a method of making a reduced metal nanoparticle, the method including mixing a reactive reducing agent with a metal salt in a solution at a temperature of 4-100° C., and forming the reduced metal nanoparticles in the solution. Also described is a kit including a reactive reducing agent that is sensitive to ßgalactosidase, a metal salt, and optionally a modifying agent/functionalizing agent for reduced metal nanoparticles. A 3,4-cyclohexeneoesculetin-B-D-galacto pyranoside (SGNP) gold nano article and its use for measuring ßgalactosidase enzyme activity, comprising by detecting a structural change in the SGNPs caused by the ßgalactosidase are described. Further described are a point of care device, a chip, a biosensor, a laboratory animal, a gene delivery agent, a drug delivery agent, a diagnostic agent, or a disease targeting agent including SGNPs.

Abstract: An RNA tagging system for visualization of single mRNA molecules based on a MSB-MCP system, as well as methods of use.

Abstract: A method for improving the performance of a fluorescence microscopy imaging system and for correcting chromatic aberration of an optical objective in a fluorescence microscopy system.

Abstract: Methods and apparatus are disclosed for imaging molecular interactions in living cells at high resolution, low light levels and high acquisition speeds.

Abstract: The present invention generally relates to methods for determining tumor resistance or sensitivity to chemotherapeutic agents and the likelihood of tumor reoccurrence based on the expression levels of genes known to correlate to the chemotherapeutic agent. In particular, the expression levels of TYMS, MRGX, ATP7B and/or BAK in tumor cells, as measured by the number of active transcription sites detected by fluorescence in situ hybridization (FISH), are predictive of resistance and sensitivity to chemotherapy and the likelihood of reoccurrence following chemotherapy treatment.

Abstract: A method for visualizing the location and movement of a specific RNA of interest in a living cell, in real time, is disclosed. The method includes the following steps: (a) providing a DNA encoding the RNA, which RNA includes a protein-binding site; (b) providing a nucleic acid encoding a fusion protein, which fusion protein comprises a fluorescent domain and an RNA-binding domain; (c) introducing the DNA encoding the RNA, and the nucleic acid encoding the fusion protein, into a eukaryotic cell so that the DNA encoding the RNA and the nucleic acid encoding the fusion protein are expressed in the cell; and (d) detecting fluorescence outside the nucleus or inside the nucleus of the cell, with the fluorescence being from the fusion protein bound to the RNA. In some embodiments, the fusion protein also includes an intracellular localization domain.

Abstract: An in situ hybridization method for detecting and specifically identifying transcription of a multiplicity of different target sequences in a cell is disclosed. The method includes assigning a different bar code to at least five target sequences, with each target sequence containing at least one predetermined subsequence. Each bar code contains at least one fluorochrome, and at least one bar code comprises at least two different, spectrally distinguishable fluorochromes. A probe set specific for each target sequence is provided in the method. Each probe set contains a hybridization probe complementary to each subsequence in the target sequence. Each probe is labeled with a fluorochrome, and the fluorochromes in each probe set collectively correspond to the bar code for the target sequence of that probe set.

Abstract: Improved methodologies for in-situ hybridization and non-isotopic detection of nucleic acid sequences are provided which offer major increases of resolution, sensitivity, and simplicity unavailable in previously known techniques. The methodology is able to detect even a single-copy of a specific nucleic acid of interest under controlled conditions regardless of whether these are DNA or RNA sequences; or whether the nucleic acid sequence of interest is localized in the chromosomes, nucleus, or cytoplasm of a cell. The methods employ a variety of non-isotopic labels and detection means for rapid and reliable assays. The invention is also provided in kit form for use in the clinical/diagnostic laboratory such that a relatively unskilled person can accurately and reproducibly detect even a single-copy of a specific nucleic acid of interest.

Abstract: A method for visualizing the location and movement of a specific RNA of interest in a living cell, in real time, is disclosed. The method includes the following steps: (a) providing a DNA encoding the RNA, which RNA includes a protein-binding site; (b) providing a nucleic acid encoding a fusion protein, which fusion protein comprises a fluorescent domain and an RNA-binding domain; (c) introducing the DNA encoding the RNA, and the nucleic acid encoding the fusion protein, into a eukaryotic cell so that the DNA encoding the RNA and the nucleic acid encoding the fusion protein are expressed in the cell; and (d) detecting fluorescence outside the nucleus or inside the nucleus of the cell, with the fluorescence being from the fusion protein bound to the RNA. In some embodiments, the fusion protein also includes an intracellular localization domain.

Abstract: Improved methodologies for in-situ hybridization and non-isotopic detection of nucleic acid sequences are provided which offer major increases of resolution, sensitivity, and simplicity unavailable in previously known techniques. The methodology is able to detect even a single-copy of a specific nucleic acid of interest under controlled conditions regardless of whether these are DNA or RNA sequences; or whether the nucleic acid sequence of interest is localized in the chromosomes, nucleus, or cytoplasm of a cell. The methods employ a variety of non-isotopic labels and detection means for rapid and reliable assays. The invention is also provided in kit form for use in the clinical/diagnostic laboratory such that a relatively unskilled person can accurately and reproducibly detect even a single-copy of a specific nucleic acid of interest.

Abstract: Disclosed is a method of detecting a trinucleotide repeat expansion by in situ hybridization. The disclosed method uses a sample of nucleated cells, a labeled trinucleotide repeat-specific probe and detection of the hybridized probe by a means whose sensitivity distinguishes between the signal from probes hybridized to an expanded repeat and the signal from probes hybridized to a non-expanded repeat.

Abstract: Disclosed are methods for accurately determining the total emission intensity of a single fluorochrome, under imaging conditions, using a digital imaging fluorescence microscopy system. Also disclosed are methods for detecting and localizing probe-target molecule binding. The detection method has sufficient resolution and sensitivity to locate and detect a single target-bound probe.

Abstract: Disclosed are methods for removing autofluorescence background from microscopic images or from flow cytometer measurements. The microscopic images can be fluorescent in situ hybridization images or images of living cells. The methods involve the steps of: (1) detecting fluorescence from a cell before "uncaging" a caged fluorochrome label on a probe; (2) uncaging the fluorochrome; (3) detecting fluorescence from the cell; and (4) subtracting fluorescence detected before uncaging from fluorescence detected after uncaging. Also disclosed is a method for tracking the movement of a target molecule in a living cell. Also disclosed is a fluorochrome-uncaging flow cytometer. The fluorochrome-uncaging flow cytometer includes a first fluorescence excitation light beam, an uncaging light beam, a second fluorescence excitation light beam, an electronic data system, a first photodetector operably linked to the electronic data system, and a second photodetector operably linked to the electronic data system.

Abstract: A method of nucleic acid hybridization in living cells is described, which is useful for detecting, quantitating and locating a specific nucleic acid in a cell or tissue, for selecting cells based on the expression or presence of a specific nucleic acid, and for monitoring the amount and location of a specific nucleic acid over time or under various inducing or inhibiting conditions.

Abstract: Disclosed are the particular 3' noncoding nucleotide sequences (zipcode and zipcode elements) that cause .beta.-actin mRNA (and .beta.-actin protein synthesis) to be localized in specific regions of the cytoplasm. Also disclosed is the discovery that zipcode function can be fully inhibited by antisense oligonucleotides.

Abstract: Strand Displacement Amplification for amplification of nucleic acid target sequences in situ in cells in suspension, on slides or in tissues. Excellent specimen morphology is preserved, and either DNA targets, or RNA targets, or both may be selectively amplified. In situ amplification by SDA is compatible with immunochemical techniques, so that both amplification of target sequences and immunological staining can be performed on the same specimen.

Abstract: Improved methodologies for in-situ hybridization and detection of hybridized nucleic acid sequences in cell cultures and tissue sections are provided which offer an increase of speed, sensitivity, and simplicity unavailable in previously known techniques. The invention detects specific nucleic acids of interest, particularly RNA sequences, within cells and tissues utilizing DNA of a particular size as a probe to find those sequences which are held substantially in common between the cell or tissue and the probe. The cells are fixed preferably in paraformaldehyde and then hybridized using a hybridization fluid for not less than 10 minutes but not substantially more than 24 hours. A variety of identifying labels are attached to the probe which permit quick and rapid detection via measurement of radioactive isotope decay or by colorimetric detection of enzymatic reaction products.