Abstract: The indicator compounds of the present invention are substituted or unsubstituted 5'-nitro-BAPTA chelators that contain a benzazolyl-coumarin substructure, and the pharmaceutically acceptable non-toxic salts and esters thereof. These compounds are useful for the detection and quantification of polycationic metal ions, particularly Zn.sup.2+, Pb.sup.2+, Ba.sup.2+, Cd.sup.2+, Hg.sup.2+ and La.sup.3+.The compounds of the invention have the structure: ##STR1## where m=2 or 3 and X can be S, O, or C(CH.sub.3).sub.2 ; optionally substituted by substituents that alter the binding affinity of the indicator, shift the spectral properties of the indicator, or act as a reactive site for the preparation of a variety of conjugates.

Abstract: The dyes of the present invention are fluorescent substituted 3',6'-diaminoxanthenes and their reduced analogs 3',6'-diaminodihydroxanthenes, which are oxidized to the fluorescent form of the dye in situ. In their oxidized form, the dyes selectively localize within mitochondria. The dyes of the present invention include an alkylating group that is covalently attached that allows their retention in mitochondria even after cell death, fixation, and permeabilization.

Abstract: The invention relates to fluorescent and/or reactive derivatives of 1,2-bis-(2-aminophenoxyethane)-N,N,N',N'-tetraacetic acid (BAPTA) according to the formula: ##STR1## where at least one of W and X is a functional group, with or without a spacer, that terminates in an alcohol or phenol, a thiol, a haloacetamide, an alkyl halide, an amine or aniline, a carboxylic acid, an anhydride, an isocyanate, an isothiocyanate, a maleimide, or an activated ester. The BAPTA-like molecule may be further substituted, one or more times, by additional functional groups with or without spacers or by CH.sub.3, NO.sub.2, CF.sub.3, F, Cl, Br, I, or carboxylic acid derivatives or pharmaceutically acceptable salts thereof, or by indolyl or benzofuran fluorophores. The functional groups allow for subsequent covalent attachment of one or more oxygen heterocycle fluorophores (e.g. fluorescein, coumarin, rhodamine); or polymolecular assemblies (e.g.

Abstract: This invention relates to derivatives of dipyrrometheneboron difluoride fluorescent dyes that have an absorption maximum at wavelengths longer than about 525 nm, and are chemically reactive with nucleic acids, proteins, carbohydrates, and other biologically derived or synthetic chemical materials to form dye-conjugates. The dye-conjugates generally have the structure: ##STR1## wherein at least one of the substituents R.sub.1 -R.sub.7, covalently bonds to a specific binding pair member, and further where at least one of the substituents R.sub.1 -R.sub.7 contains a bathochromic moiety that is heteroaryl or alkenyl. The remaining substituents, which may be the same or different, are hydrogen, halogen, alkyl (containing 1-5 carbon atoms), aryl, arylalkyl, or sulfo.

Abstract: The invention relates to a method of forming distinctive intravacuolar structures that are unique to fungal cells and can be correlated with cell viability. The preferred dyes of the present invention are described by the formula: ##STR1## wherein each R.sup.1 is independently H; or an alkyl group having from 1-6 carbons; or a trifluoromethyl; or a halogen; or --OR.sup.6, --SR.sup.6 or --(NR.sup.6 R.sup.7) where R.sup.6 and R.sup.7, which can be the same or different, are independently H; or alkyl groups having 1-6 carbons; or 1-2 alicyclic, heteroalicyclic, aromatic or heteroaromatic rings, containing 1-4 heteroatoms, wherein the heteroatoms are O, N or S; or R.sup.6 and R.sup.7 taken in combination are --(CH.sub.2).sub.2 --M--(CH.sub.2).sub.2 -- where M=a single bond, --O--, --CH.sub.2 --, or --NR.sup.8 -- where R.sup.8 is H or an alkyl having 1-6 carbons; and t=1-4;R.sup.2 is an alkyl group having 1-6 carbons;X is O, S, Se or NR.sup.9, where R.sup.9 is H or an alkyl group having 1-6 carbons; or X is CR.

Abstract: Activity of enzymes and enzyme conjugates is detected using novel substrates made from a class of fluorophores, generally including quinazolinones (quinazolones), benzimidazoles, benzothiazoles, benzoxazoles, quinolines, indolines, and phenanthridines, having the general formula: ##STR1## where carbon atoms of --C.sup.1 .dbd.C.sup.2 -- are joined to complete a first 5- or 6-membered aromatic ring which optionally contains one of the hetero atoms N, O or S,and carbon atoms of --C.sup.4 --N.dbd.C.sup.3 -- are joined to complete a second 5- or 6-membered aromatic ring that contains a nitrogen between C.sup.3 and C.sup.4 and optionally contains an additional hetero atom N, O or S,where the first and second aromatic rings may be joined by a 5- or 6-membered bridging ring that contains at least the C.sup.2 from the first aromatic ring and the C.sup.

Abstract: Novel fluorescent dyes based on the rhodol structure are provided. The new reagents contain functional groups capable of forming a stable fluorescent product with functional groups typically found in biomolecules or polymers including amines, phenols, thiols, acids, aldehydes and ketones. Reactive groups in the rhodol dyes include activated esters, isothiocyanates, amines, hydrazines, halides, acids, azides, maleimides, aldehydes, alcohols, acrylamides and haloacetamides. The products are detected by their absorbance or fluorescence properties. The spectral properties of the fluorescent dyes are sufficiently similar in wavelengths and intensity to fluorescein or rhodamine derivatives as to permit use of the same equipment. The dyes, however, show less spectral sensitivity to pH in the physiological range than does fluorescein, have higher solubility in non-polar solvents and have improved photostability and quantum yields.

Abstract: The invention relates to use of fluorescent compounds of the formula: ##STR1## where R contains between 4 and about 10 carbons and is optionally saturated or unsaturated, and is linear or branched or contains an alicyclic or aromatic ring; and the symbol .PSI. depicts the presence of the counterion used to neutralize the positive charge on the dye. The fluorescent dye dissolved in a biologically compatible solution stains a wide variety of living cells with a red nucleic acid stain after brief incubation in low concentrations of dye, without the requirement of permeabilizing reagents. Detection of the fluorescence can be used alone or in combination with measurement of other markers or properties of the cells to identify, discriminate or sort viable cells.

Abstract: The invention describes the preparation and use of fluorescent stains for nucleic acids derived from unsymmetrical cyanine dyes. In particular, the invention describes unsymmetrical cyanine dyes having a saturated or unsaturated cyclic substituent. The dyes of the invention possess superior fluorescent characteristics when complexed with nucleic acids, and have utility in any application which requires detection of nucleic acids. The presence of the cyclic substituent results in improved permeability in a wide range of living cells, resulting in improved detection of intracellular nucleic acids.

Abstract: The invention relates to fluorescent dyes that are substituted or unsubstituted derivatives of 1-[isoindolyl]methylene-isoindole that are bound through both isoindole nitrogens to a boron difluoride moiety, forming a fluorescent dibenzopyrrometheneboron difluoride compound ##STR1## whose fluorescence properties are modified by the selection of appropriate chemical substituents.The dibenzopyrrometheneboron difluoride compound is optionally substituted by hydrogen, halogen cyano, sulfo, alkali or ammonium salts of sulfo, carboxy, substituted or unsubstituted alkyl, perfluoroalkyl, alkoxy, alkylthio, nitro, amino, monoalkylamino, dialkylamino, substituted or unsubstituted aryl substituents, substituted or unsubstituted heteroaryl substituents, or additional substituted or unsubstituted fused benzo rings or substituted or unsubstituted fused heteroaromatic rings.

Abstract: The invention relates to nucleic acid stains that are homo- or heterodimers of unsymmetrical cyanine dyes, where at least one dye unit of the dimer is a substituted or unsubstituted benzazolium- or naphthazolium-pyridinium derivative, which dye unit is linked to another unsymmetrical cyanine dye unit, that is the same or different by an aliphatic chain containing one or more heteroatoms; such that the fluorescence of the dye molecules, when bound to nucleic acids, is enhanced. These dimers of unsymmetrical cyanine dyes have spectral properties and binding affinities superior to related monomers. In addition, the dyes can be tailored to absorb and emit in a wide range of wavelengths, which can be further varied by energy transfer mixtures, making the dyes useful for applications that involve simultaneous use of multiple dyes.

Abstract: This invention describes novel sensors for ions that are based on the combination of xanthylium-based dyes with metal-binding N,N'-diaryldiaza crown ethers. These sensors are primarily useful for detection and quantitation of alkali-metal ions in aqueous solution. Binding of the ion results in a change in the fluorescence properties of the indicating dye that can be correlated with the ion concentration. Methods are provided for attaching reactive groups on these sensors for conjugation to dyes, lipids and polymers and for enhancing entry of the indicators into living cells.

Abstract: Fluorescent compounds useful in the determination of chloramphenicol acetyltransferase (CAT) enzyme activity are described. The compounds are "fluorescent derivatives related in structure to chloramphenicol and are acylated in the presence of CAT to produce fluorescent mono- and diacylated products, which are then physically separated from the reaction mixture and quantitated by means of their fluorescence and/or absorbance. Fluorescent molecules conjugated to chloramphenicol include derivatives of fluorescein, rhodamine, coumarin, dimethylaminonaph-thalene sulfonic acid (dansyl), pyrene, anthracene, nitrobenz-oxadiazole (NBD), acridine and dipyrrometheneboron difluoride.

Abstract: The subject invention provides a method for analyzing the metabolic activity in cells by improving the retention of a detectable reporter molecule only in intact cells where a particular enzyme is present. In particular, improved retention results from a two part process involving conjugation of haloalkyl-substituted derivatives of a reporter molecule with intracellular cysteine-containing peptides while unblocking the reporter molecule. The method for analyzing metabolic activity of cells involves the use of a substrate having the formXR-REPORTER-BLOCKwherein -BLOCK is a group selected to be removable by action of a specific analyte, to give REPORTER spectral properties different from those of the substrate,-REPORTER- is a molecule that, when no longer bound to BLOCK by a BLOCK-REPORTER bond, has spectral properities different from those of the substrate, andXR-- is a haloalkyl moiety that can covalently react with an intracellular thiol (Z--S--H) to form a thioether conjugate (Z--S--R--).

Abstract: This invention relates to improved fluorescent fatty acid analogs derived from dipyrrometheneboron difluoride ("BDY") dyes that absorb light at wavelengths longer than 480 nm. The BDY fluorophore or a derivative thereof is incorporated at various locations along the alkyl portion of the fatty acid ("BDY fatty acid"). The general formula of a BDY fatty acid is as follows: ##STR1## wherein at least one of the substituents R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6 and R.sub.7 is a carboxylic acid terminated alkyl residue containing 5 to 22 carbon atoms. The remaining substituents, singly or in any combination, are: additional alkyl or arylalkyl residues containing 1 to 16 aliphatic carbon atoms, or aryl or heteroaryl residues, or hydrogen. Symmetrically substituted BDY fluorophores are conveniently synthesized from a single pyrrole precursor in a "one-pot" reaction. Alternatively, BDY fatty acids are synthesized from two different pyrrole precursors.

Abstract: The invention relates to microparticles incorporating a series of two or more fluorescent dyes having overlapping excitation and emission spectra allowing efficient energy transfer from the excitation wavelength of the first dye in the series, transfer through the dyes in the series and re-emitted as an optical signal at the emission wavelength of last dye in the series, resulting in a desired effective Stokes shift which is controlled through selection of appropriate dyes.

Abstract: The invention relates to unsymmetrical cyanine dyes with a cationic side chain, typically benzothiazole or benzoxazole derivatives, that exhibit enhanced fluorescence on binding with DNA or RNA, where such fluorescence can be used for evaluating the presence of nucleic acid polymers. The dyes generally have the formula: ##STR1## where R.sup.1 is an alkyl group having 1-6 carbons; X is 0, S, or NR.sup.2, where R.sup.2 is an alkyl group having 1-6 carbons, or CR.sup.3 R.sup.4, where R.sup.3 and R.sup.4, which may be the same or different, are alkyl groups having 1-6 carbons;n=0, 1, or 2;Z.sup.1 and Z.sup.2, which may be the same or different, are independently hydrogen, an alkyl group having 1-6 carbons, or aryl, or Z.sup.1 and Z.sup.2 taken in combination complete a 6-membered aromatic ring;Y is HC.dbd.

Abstract: Novel fluorescent precipitating substrates made from a class of fluorophores, generally including quinazolinones (quinazolones), benzimidazoles, benzothiazoles, benzoxazoles, quinolines, indolines, and phenanthridines, having the general formula: ##STR1## where carbon atoms of --C.sup.1 .dbd.C.sup.2 -- are further joined so as to complete a first 5- or 6-membered aromatic ring which may contain at least one of the hetero atoms N, O or S,where carbon atoms of --C.sup.4 --N.dbd.C.sup.3 -- are further joined so as to complete a second 5- or 6-membered aromatic ring that contains at least the nitrogen between C.sup.3 and C.sup.4 and may contain at least one additional hetero atom N, O or S,where the first and second aromatic rings may be joined by a 5- or 6-membered bridging ring that contains at least the C.sup.2 from the first aromatic ring and the C.sup.

Abstract: This invention relates to a method for simultaneously detecting live and dead cells using two fluorogenic reagents: calcein AM and ethidium homodimer. Live cells are distinguished by an intense uniform green fluorescence generated by the enzymatic hydrolysis of calcein AM: ##STR1## Dead or damaged cells are distinguished by a bright red fluorescence resulting from nucleic acids stained with ethidium homodimer: ##STR2## The assay is useful to determine cell viability and to monitor cytotoxicity agents or events.

Abstract: This invention relates to derivatives of dipyrrometheneboron difluoride fluorescent dyes that have an absorption maximum at wavelengths longer than about 525 nm, and are chemically reactive with nucleic acids, proteins, carbohydrates, and other biologically derived or synthetic chemical materials. The dyes generally have the structure: ##STR1## wherein at least one of the substituents R.sub.1 -R.sub.7, is a reactive functional group, and at least one of the substituents R.sub.1 -R.sub.7 contains a bathochromic moiety. The bathochromic moiety is an unsaturated organic group, preferably heteroaryl or alkenyl. The remaining substituents, which may be the same or different, are hydrogen, halogen, alkyl (containing 1-5 carbon atoms), aryl, arylalkyl, or sulfo. The dyes are used to make novel conjugates with members of specific binding pairs that are ligands or receptors.