Abstract: The present invention relates to novel fluorescent dyes, novel fluorogenic and fluorescent reporter molecules and new enzyme assay processes that can be used to detect the activity of caspases and other enzymes involved in apoptosis in whole cells, cell lines and tissue samples derived from any living organism or organ. The reporter molecules and assay processes can be used in drug screening procedures to identify compounds which act as inhibitors or inducers of the caspase cascade in whole cells or tissues. The reagents and assays described herein are also useful for determining the chemosensitivity of human cancer cells to treatment with chemotherapeutic drugs. The present invention also relates to novel fluorogenic and fluorescent reporter molecules and new enzyme assay processes that can be used to detect the activity of type 2 methionine aminopeptidase, HIV protease, adenovirus protease, HSV-1 protease, HCMV protease and HCV protease.

Abstract: A class of asymmetric monobenzoxanthene compounds useful as fluorescent dyes are disclosed having the structure wherein Y1 and Y2 are individually hydroxyl, amino, imminium, or oxygen, R1-R8 are hydrogen, fluorine, chlorine, alkyl, alkene, alkyne, sulfonate, amino, amido, nitrile, alkoxy, linking group, and combinations thereof and R9 is acetylene, alkane, alkene, cyano, substituted phenyl, and combinations thereof. The invention further includes novel intermediate compounds useful for the synthesis of asymmetric benzoxanthene compounds having the general structure where substituents R3-R7 correspond to like-referenced substituents in the structure of described above, and Y2 is hydroxyl or amine. In another aspect the invention includes methods for synthesizing the above dye compounds and intermediates.

Abstract: A compound represented by general formula (IA) or a salt thereof useful as a fluorescent probe for zinc: wherein R1 and R2 represent a hydrogen atom or a group represented by formula (A), wherein X1, X2, X3, and X4 represent a hydrogen atom, an alkyl group, a 2-pyridylmethyl group, or a protective group for an amino group, and m and n represent 0 or 1 provided that R1 and R2 do not simultaneously represent hydrogen atoms; R3 and R4 represent a hydrogen atom or a halogen atom; and R5 and R6 represent a hydrogen atom, an alkylcarbonyl group, or an alkylcarbonyloxymethyl group, and R7 represents a hydrogen atom or an alkyl group.

Abstract: A compound represented by general formula (I) or (II) or a salt thereof and an agent for measurement of a reactive oxygen comprising said compound or a salt thereof: 1

Abstract: A sensitizer for tumor treatment has a structure such that a lipophilic group and a hydrophilic group are bound to one and the same carbon atom in a compound containing a xanthene dye structure. It is important that the lipophilic group and hydrophilic group are bound to one and the same carbon atom; if they are separated from each other, no sufficient effect is obtained. Thus, the sensitizer possesses both lipophilicity and hydrophilicity. This sensitizer has a function of promoting acoustic cavitation and a function of revealing an antitumor effect and is an agent highly accumulable in tumor tissues. Thus, this sensitizer is highly selective for tumor tissues and enables the treatment of malignant tumor using ultrasound.

Abstract: Fluorescein derivative as a substrate for cytochrome P450 enzymes.

Abstract: The subject matter of the invention are new xanthene derivatives of the formula I, wherein R1,R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, and X, Y, are as defined herein. The compounds according to the invention provide molecules that are—due to their spectral properties (absorption maxima in the range of approx. 650 nm and above as well as emission maxima above 670 nm)—very suitable for the use as dyes and in particular as fluorescence dyes. The compounds of the formula I according to the invention are used for the production of fluorescence conjugates, for their application in immunoassays, for DNA analytics, for in-vivo diagnostics or as a laser dye.

Abstract: Atropisomeric energy-transfer dye compounds are disclosed. A variety of molecular biology applications utilize atropisomeric xanthene fluorescent dyes as labels for substrates such as nucleotides, nucleosides, polynucleotides, polypeptides and carbohydrates. Methods include DNA sequencing, DNA fragment analysis, PCR, SNP analysis, oligonucleotide ligation, amplification, minisequencing, and primer extension.

Abstract: Disclosed are compounds having two kinds of reporters that can be a donor and an acceptor for energy transfer, for example, fluorescent groups, and having a 2′, 3′-dideoxyribonucleotide residue or a 3′-deoxyribonucleotide residue. These compounds can be used as terminators for the chain terminator method. The two kinds of reporters are arranged with a distance sufficient for causing energy transfer from the donor to the acceptor. Also disclosed are methods for determining DNA sequences based on the chain terminator method wherein the chain termination reaction is performed by using the above terminators. Also disclosed are compounds having two kinds of reporters that can be a donor and an acceptor for energy transfer, which can be used as a primer or an initiator in methods for determining DNA sequences utilizing the chain terminator method, and methods for determining DNA sequences utilizing the compounds.

Abstract: Electrophoretic probes comprising fluorescent compounds as detection groups and mobility modifiers are disclosed for the multiplexed detection of the binding of, or interaction between, one or more ligands and target antiligands are provided. In one embodiment, detection involves the release of identifying tags as a consequence of target recognition. Target antiligands are contacted with a set of e-tag probes and the contacted antiligands are treated with a selected cleaving agent resulting in a mixture of e-tag reporters. Typically, uncleaved or partially cleaved e-tag probes are removed and the mixture of e-tag reporters is separated by any technique that provides for separation by mass or mass to charge ratio and the like and detected to provide for target identification.

Abstract: Substantially pure atropisomers of xanthene compounds are disclosed. A variety of molecular biology applications utilize atropisomeric xanthene fluorescent dyes as labels for substrates such as nucleotides, nucleosides, polynucleotides, polypeptides and carbohydrates. Methods include DNA sequencing, DNA fragment analysis, PCR, SNP analysis, oligonucleotide ligation, amplification, minisequencing, and primer extension.

Abstract: The present invention is directed, in part, to fluorescein-based ligands for detection of metal ions, and methods of making and using the same.

Abstract: Process for the preparation of compounds of formula (I), wherein the general symbols are as defined in claim 1, which process comprises reacting a compound of formula (V), wherein the general symbols are as defined in claim 1, with carbon monoxide in the presence of a catalyst.

Abstract: Long wavelength, narrow emission bandwidth fluorecein dyes are provided for detecting spacially overlapping target substances. The dyes comprise 4,7-dichlorofluoresceins, and particularly 2′,4′,5′,7′-tetrachloro-4,7-dicloro-5- (and 6-) carboxyfluoresceins. Methods and kits for using the dyes in DNA analysis are provided.

Abstract: The present invention provides novel, water-soluble, red-emitting fluorescent rhodamine dyes and red-emitting fluorescent energy-transfer dye pairs, as well as labeled conjugates comprising the same and methods for their use. The dyes, energy-transfer dye pairs and labeled conjugates are useful in a variety of aqueous-based applications, particularly in assays involving staining of cells, protein binding, and/or analysis of nucleic acids, such as hybridization assays and nucleic acid sequencing.

Abstract: Fluorescent energy transfer cassettes that allow through bond energy transfer and have a succinimidyl ester functionality suitable for affecting them to biomolecules or provided and are applied to high throughput DNA sequencing.

Abstract: A hydrocarbyldithiomethyl-modified compound of the Formula:

Abstract: Immunoassay reagents, methods and test kits for the specific quantification of vancomycin in a test sample are disclosed. The reagent comprises antibodies prepared with immunogens of FIG. 6 wherein P is an immunogenic carrier material and X is a linking moiety.

Abstract: A sensitizer for tumor treatment has a structure such that a lipophilic group and a hydrophilic group are bound to one and the same carbon atom in a compound containing a xanthene dye structure. It is important that the lipophilic group and hydrophilic group are bound to one and the same carbon atom; if they are separated from each other, no sufficient effect is obtained. Thus, the sensitizer possesses both lipophilicity and hydrophilicity. This sensitizer has a function of promoting acoustic cavitation and a function of revealing an antitumor effect and is an agent highly accumulable in tumor tissues. Thus, this sensitizer is highly selective for tumor tissues and enables the treatment of malignant tumor using ultrasound.

Abstract: A class of asymmetric monobenzoxanthene compounds useful as fluorescent dyes are disclosed having the structure wherein Y1 and Y2 are individually hydroxyl, amino, imminium, or oxygen, R1-R8 are hydrogen, fluorine, chlorine, alkyl, alkene, alkyne, sulfonate, amino, amido, nitrile, alkoxy, linking group, and combinations thereof, and R9 is acetylene, alkane, alkene, cyano, substituted phenyl, and combinations thereof. The invention further includes novel intermediate compounds useful for the synthesis of asymmetric benzoxanthene compounds having the general structure where substituents R3-R7 correspond to like-referenced substituents in the structure of described above, and Y2 is hydroxyl or amine. In another aspect, the invention includes methods for synthesizing the above dye compounds and intermediates.

Abstract: The present invention provides a 13C-labeled oligosaccharide or polysaccharide or a salt thereof excluding U-13C-maltose, 13C-starch, 1-13C-maltotetraose and 1-13C-amylose; a derivative of the 13C-labeled oligosaccharide or polysaccharide or salt thereof; a 13C-labeled inclusion complex or a salt thereof, which comprises a cyclodextrin or a modified derivative thereof as a host molecule; a 13C- or 14C-labeled fluorescein ester compound or a salt thereof; and a diagnostic agents for pancreatic exocrine function comprising these compounds 13C- or 14C-labeled.

Abstract: An in vitro assay method permits the identification of a test substance which inhibits the mutual association of two molecules. The method includes the steps of providing two components capable of mutual association, one of said components bearing a covalently linked fluorophore; preparing a mixture containing the two components and at least one test substance; irradiating the mixture with polarized light of a suitable wavelength permitting excitation of the fluorophore as indicated by emission of polarized light; measuring the degree of polarization of the emission, and determining the effect of the presence or concentration of the test substance in decreasing the observed emission polarization of a mixture of the two components alone. Inhibitory activity of the test substance correlates with decreased depolarization values.

Abstract: A micro-circuit for performing analyses of multimolecular interactions and for performing molecular syntheses, comprising: (a) a support; (b) at least one micro-electrode attached to the support, the micro-electrode being selectively electronically activated and the micro-electrode having a protective layer which is removable; (c) a binding entity for attachment to the at least one micro-electrode, the binding entity being capable of attachment to at least one micro-electrode when the protective layer has been removed; and (d) a power source being operatively connected to at least one micro-electrode for electronically activating at least one micro-electrode. The micro-circuit of the present invention also includes embodiments featuring a micro-circuit reader for detecting the interaction of the binding entity to a complementary probe, as well as methods for making and using the micro-circuit of the present invention.

Abstract: The present invention provides novel, water-soluble, red-emitting fluorescent rhodamine dyes and red-emitting fluorescent energy-transfer dye pairs, as well as labeled conjugates comprising the same and methods for their use. The dyes, energy-transfer dye pairs and labeled conjugates are useful in a variety of aqueous-based applications, particularly in assays involving staining of cells, protein binding, and/or analysis of nucleic acids, such as hybridization assays and nucleic acid sequencing.

Abstract: A double- or single-stranded oligonucleotide comprising one or more respectively inter- or intra-oligonucleotide covalent cross-linkages, wherein the or each covalent linkage consists of an amide bond between a primary amine group of one strand and a carboxyl group of the other strand or the same strand, respectively, said primary amine group being directly substituted in the 2′ position of the strand nucleotide monosaccharide ring, and said carboxyl group being carried by an aliphatic spacer group substituted on a nucleotide or nucleotide analog of the other strand or the same strand, respectively. A method for preparing such oligonucleotides, and nucleotide or non-nucleotide synthons useful for preparing an oligonucleotide of the above kind, are also disclosed.

Abstract: Xanthan esters and acridans are substrates for horseradish peroxidase. These stable, enzymatically cleavable chemiluminescent esters are substrates for horseradish peroxidase which, together with peroxide is among the extensively used enzyme in enzyme-linked detection methods, including immunoassays, oligonucleotide detection and nucleic acid hybridization. The novel compounds are used, together with peroxide, alkali and the peroxidase, to indicate the presence and/or concentration of target compounds. The assays may be enhanced by the use of polymeric quaternary onium enhancement compounds or similar compounds selected to enhance the chemiluminescence emitted.

Abstract: The family of dyes of the invention are fluoresceins and rhodols that are directly substituted on one or more aromatic carbons by fluorine. These fluorine-substituted fluorescent dyes possess greater photostability and have lower sensitivity to pH changes in the physiological range of 6-8 than do non-fluorinated dyes, exhibit less quenching when conjugated to a substance, and possess additional advantages. The dyes of the invention are useful as detectable tracers and for preparing conjugates of organic and inorganic substances.

Abstract: Lipophilic fluorescein-based molecules have been synthesized and incorporated into thin plasticized polymeric membrane films as chromoionophores for the optical sensing and detection of polycationic protamine. The membrane response is based on the extraction of protamine into the film due to the interaction between the anionic fluorescein and the protamine polycation which results in a cation exchange between the protamine and proton, and thus, induces an absorbance spectra change of the polymeric film. The response speed is controlled by protamine diffusion through the stagnant diffusion layer adjacent to the film surface as well as within the bulk of the polymer film. When limited exposure time and non-stirring detection modes are used in a buffer solution, absorbance of a film changes as a function of the protamine concentration in the range of about 2 to 60 .mu.g/ml (0.44 to 13.3 .mu.M).

Abstract: Provided is an electron donating dye precursor represented by the following formula (1), (2), (3) or (4) as defined in the specification which each singly develops two colors upon color forming reaction with one kind of an electron accepting color developer depending on change in the heat energy applied. Furthermore, a heat-sensitive recording material comprising a support and a recording layer containing said electron donating dye precursor and an electron accepting color developer is provided. Moreover, a method for heat sensitive recording of multi-color images using said heat-sensitive recording material is provided.

Abstract: The present invention describes xanthene dyes, including rhodamines, rhodols and fluoresceins that are substituted one or more times by a sulfonic acid or a salt of a sulfonic acid. The dyes of the invention, including chemically reactive dyes and dye-conjugates are useful as fluorescent probes, particularly in biological samples.

Abstract: A class of asymmetric monobenzoxanthene compounds useful as fluorescent dyes are disclosed having the structure ##STR1## wherein Y.sub.1 and Y.sub.2 are individually hydroxyl amino, imminium, or oxygen, R.sub.1 -R.sub.8 are hydrogen, fluorine, chlorine, alkyl, alkene, alkyne, sulfonate, amino, amido, nitrile, alkoxy, linking group, and combinations thereof, and R.sub.9 is acetylene, alkane, alkene, cyano, substituted phenyl, and combinations thereof. The invention further includes novel intermediate compounds useful for the synthesis of asymmetric benzoxanthene compounds having the general structure ##STR2## where substituents R.sub.3 -R.sub.7 correspond to like-referenced substituents in the structure of described above, and Y.sub.2 is hydroxyl or amine. In another aspect, the invention includes methods for synthesizing the above dye compounds and intermediates.

Abstract: The present invention relates to a novel class of tracer compounds for fluorescence polarization imunoassays. The novel 4'-methyl fluorescein derivatives are conjugated to ligands via cyclic linkers.

Abstract: Therapeutic compositions for the treatment of infectious diseases are disclosed. These compositions comprise xanthones and xanthone derivatives, such as 2,3,4,5,6-pentahydroxyxanthone. Also disclosed are methods for the treatment of infectious diseases using such compounds.

Abstract: Novel linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye are provided. These linkers faciliate the efficient transfer of energy between a donor and acceptor dye in an energy transfer dye. One of these linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye has the general structure R.sub.21 Z.sub.1 C(O)R.sub.22 R.sub.28 where R.sub.21 is a C.sub.1-5 alkyl attached to the donor dye, C(O) is a carbonyl group, Z.sub.1 is either NH, sulfur or oxygen, R.sub.22 is a substituent which includes an alkene, diene, alkyne, a five and six membered ring having at least one unsaturated bond or a fused ring structure which is attached to the carbonyl carbon, and R.sub.28 includes a functional group which attaches the linker to the acceptor dye.

Abstract: A compound useful for the measurement of nitrogen monoxide under a substantially neutral condition which is represented by the following formula (I): ##STR1## wherein R.sup.1 and R.sup.2 represent amino groups that substitute at adjacent positions on the phenyl ring; R.sup.3 and R.sup.4 independently represent a hydrogen atom or an acyl group; R.sup.5, R.sup.6, R.sup.7, and R.sup.8 independently represent a hydrogen atom, a C.sub.1-6 alkyl group, allyl group, or a halogen atom.

Abstract: Energy transfer fluorescent dyes, reagents incorporating the dyes, kits and methods for using the dyes and reagents are provided. The energy transfer fluorescent dyes include a donor dye which absorbs light at a first wavelength and emits excitation energy in response, the donor dye including a xanthene ring structure having a 4' ring position, an acceptor dye capable of absorbing the excitation energy emitted by the donor dye and fluorescing at a second wavelength in response, and a linker attaching the donor dye to the acceptor dye, the linker having a 4' end which includes a R.sub.1 XC(O)R.sub.2 group where R.sub.1 is a C.sub.1-5 alkyl attached to the 4' ring position of the donor dye, X selected from the group consisting of NH, sulfur and oxygen, C(O) is a carbonyl group, and R.sub.2 includes an alkene, diene, alkyne, a five and six membered ring having at least one unsaturated bond or a fused ring structure which is attached to the carbonyl carbon.

Abstract: Method for the rapid diagnostic of an infection of the urinary tract (UTI), by a specific examination of urine.The method of analyzing a urine specimen, for the rapid and simultaneous counting of both viable bacteria and leucocytes (white blood cells or WBC), consists of:(1) simultaneous labeling of said bacteria and said white blood cells (WBC) present in said urine specimen, with a fluorescent viability marker of bacteria, said marker tagging and accumulating in both bacteria and white blood cells, under the same conditions and leading to different fluorescent elements;(2) automatically counting all the fluorescent elements by photoelectric detection of the fluorescent marker; and(3) simultaneously classifying said fluorescent elements in fluorescent viable bacteria and in fluorescent white blood cells, in view to obtain in one part the total number of bacteria and on the other part, the total number of white blood cells, on the basis of their size.

Abstract: Dioxetanes which couple with organic and biological molecules of the formula: ##STR1## wherein X is a leaving group which is removed by an activating agent to produce light, wherein A is a coupling substituent, Ar is a substituent selected from the group consisting of phenyl and naphthyl to provide a label are described. R.sub.1 is an optional linker substituent and can have between 1 and 30 carbon atoms with some of the carbon atoms being oxygen, sulfur, nitrogen or phosphorus. Ar as phenyl is preferred. The dioxetane coupled molecules are useful in assays of all types where luminescence can be used as an indicator.

Abstract: The subject invention describes compounds containing a polyhalogenated aryl moiety. The compounds of the invention are particularly useful for the assay of a variety of enzymes, including intracellular enzymes. The subject invention also describes assays for glutathione and/or glutathione transferase enzymes. Selected compounds of the invention are particularly useful for improving the retention of fluorescent products of enzyme metabolism in cells.

Abstract: 5(6)-methyl substituted fluorescein derivatives and a process for producing 5(6)-methyl substituted derivatives. Also provided are methods for these utilizing these derivatives as indicator reagents in assays for analytes, indicator reagents which comprise specific binding members attached to these derivatives and test kits which contain these derivatives.

Abstract: The present invention provides specific binding solid phase assay methods and kits for the detection of the presence or absence of a microorganism by directly staining the microorganism and specifically capturing the stained microorganism on a solid support. The methods find particular utility in the detection of Candida. The methods may simultaneously detect the presence or absence of multiple microorganisms.

Abstract: Compounds useful to attach a fluorescein label to an oligonucleotide are disclosed. To create these compounds, oxygen groups on the fluorescein moiety are protected (e.g. with acyl groups). One then links a phosphoramidite to the fluorescein moiety via an amide or thiourea linkage to an active site on the fluorescein ring. The resulting compound can then be directly linked to an oligonucleotide as it is being formed in an automated synthesizer and then deprotected using the same deprotection conditions as are used to deprotect the oligonucleotides. Such compounds are, inter alia, useful to create labelled primers for DNA sequencing.

Abstract: The present invention relates to novel N-substituted azaheterocyclic carboxylic acids and esters thereof in which a substituted alkyl chain forms part of the N-substituent or salts therefor, to methods for their preparation, to compositions containing them, and to their use for the clinical treatment of painful, hyperalgesic and/or inflammatory conditions in which C-fibers play a pathophysiological role by eliciting neurogenic pain or inflammation.

Abstract: Long wavelength, narrow emission bandwidth fluorecein dyes are provided for detecting spacially overlapping target substances. The dyes comprise 4,7-dichlorofluoresceins, and particularly 2',4',5',7'-tetrachloro-4,7-dichloro-5- (and 6-) carboxyfluoresceins. Methods and kits for using the dyes in DNA analysis are provided.

Abstract: Immunoassay methods and reagents for the specific quantification of thyroxine in a test sample are disclosed employing antibodies prepared with thyroxine derivatives of the formula: ##STR1## wherein P is an immunogenic carrier material and X is a linking moiety. The present invention also describes the synthesis of unique labelled reagent of the formula: ##STR2## wherein Q is a detectable moiety and W is a linking moiety, preferably fluorescein or a fluorescein derivative.

Abstract: A photochromic compound, in particular, for tinting optical elements made of a plastic material has reduced mobility in plastic material. The photochromic compound is a photochromic 2H-pyran and is distinguished by the aromatic residues being chemically rigidly attached in the 2-position to a spiropyran.

Abstract: A process for the manufacture of fluorescein compounds suitable for conversion to FDA certified dyes is described. The fluorescein compounds, and in particular fluorescein (also known as Solvent Yellow 94 or D&C Yellow No.7) and 3,4,5,6-tetrachlorofluorescein, are made by a process involving the condensation of a phthalic anhydride with several molar excess of a resorcinol, the resorcinol being employed not only as a reactant, but also as a solvent for the reaction. This process also can employ the unreacted resorcinol and any intermediates or by-products from the condensation reaction as a reactant and as a solvent in subsequent condensation reactions.

Abstract: 5(6)-methyl substituted fluorescein derivatives and a process for producing 5(6)-methyl substituted derivatives. Also provided are methods for these utilizing these derivatives as indicator reagents in assays for analytes, indicator reagents which comprise specific binding members attached to these derivatives and test kits which contain these derivatives.

Abstract: Triggerable dioxetanes with a fluorescent substituent bonded or tethered in the dioxetane so as to produce fluorescence from the group are described. The compounds are of the formula: ##STR1## wherein R.sub.1, R.sub.2 and R.sub.3 are carbon containing groups and wherein one of R.sub.1, R.sub.2 and R.sub.3 is a tethered fluorescent substitutent. The compounds are useful in immunoassays and in probes using enzymes or other chemicals for triggering the dioxetanes to produce light from the fluorescent molecule in the group as a signal.

Abstract: Immunoassay methods and reagents for the specific quantification of amitriptyline or nortriptyline in a test sample are disclosed employing antibodies prepared with amitriptyline or nortriptyline derivatives of the Formula III: ##STR1## wherein for amitriptyline, R is CH.sub.3, and for nortriptyline, R is H. The present invention also describes the synthesis of unique fluorescein tracers of the structure of Formula IV and Formula V: ##STR2## wherein for a specific amitriptyline immunoassay, W.sub.1 is a heteroatom linked to the aromatic ring at the 2 or 3 position, and for a specific nortriptyline immunoassay, W.sub.2 is two heteroatoms linked together and attached to the aromatic ring at the 2 or 3 position, and wherein Q is a detectable moiety, preferably fluorescein or a fluorescein derivative.