ANTIBODY-NANOPARTICLE CONJUGATES AND METHODS FOR MAKING AND USING SUCH CONJUGATES

Disclosed herein are antibody-nanoparticle conjugates that include two or more nanoparticles (such as gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof) directly linked to an antibody or fragment thereof through a metal-thiol bond. Methods of making the antibody-nanoparticle conjugates disclosed herein include reacting an arylphosphine-nanoparticle composite with a reduced antibody to produce an antibody-nanoparticle conjugate. Also disclosed herein are methods for detecting a target molecule in a sample that include using an antibody-nanoparticle conjugate (such as the antibody-nanoparticle conjugates described herein) and kits for detecting target molecules utilizing the methods disclosed herein. 155-. (canceled)56. A kit for detecting a target molecule in a sample , comprising one or more containers comprising a first antibody conjugated to two or more nanoparticles a composition comprising a first antibody-nanoparticle conjugate , the first antibody-nanoparticle conjugate comprising three or more metal nanoparticles directly linked to a single antibody through three or more metal-thiol bonds , wherein the metal-thiol bonds are between thiol from cysteine residues of the antibody and the three or more metal nanoparticle without an intervening linker.57. The kit of claim 56 , further comprising a second antibody conjugated to one or more enzyme molecules claim 56 , wherein the first antibody specifically binds to the second antibody.58. The kit of claim 56 , wherein the two or more nanoparticles comprise gold claim 56 , palladium claim 56 , platinum claim 56 , silver claim 56 , copper claim 56 , nickel claim 56 , cobalt claim 56 , iridium claim 56 , or an alloy of two or more thereof.59. The kit of claim 56 , wherein the two or more nanoparticles are about 5 nm or less in diameter.60. The kit of claim 59 , wherein the two or more nanoparticles are about 0.5-5 nm in diameter.61. The kit of claim 56 , wherein the ...

Polymer stabilization of chromogen solutions

Disclosed embodiments concern a composition comprising DAB chromogen, and/or derivative thereof, a stabilizer, and polymer capable of preventing or reducing DAB precipitation relative to a composition that does not comprise the polymer. Also disclosed herein is a method for using the disclosed composition and embodiments of a kit.

Antibody-nanoparticle conjugates and methods for making and using such conjugates

Disclosed herein are antibody-nanoparticle conjugates that include two or more nanoparticles (such as gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof) directly linked to an antibody or fragment thereof through a metal-thiol bond. Methods of making the antibody-nanoparticle conjugates disclosed herein include reacting an arylphosphine-nanoparticle composite with a reduced antibody to produce an antibody-nanoparticle conjugate. Also disclosed herein are methods for detecting a target molecule in a sample that include using an antibody-nanoparticle conjugate (such as the antibody-nanoparticle conjugates described herein) and kits for detecting target molecules utilizing the methods disclosed herein.

ENHANCED DEPOSITION OF CHROMOGENS UTILIZING PYRIMIDINE ANALOGS

This disclosure relates to compositions that enhance the deposition of detectable moieties on tissue samples, methods utilizing these compositions and kits including these compositions. The compositions include a deposition enhancer having a formula 2. The method of claim 1 , wherein contacting the sample with a detection solution includes enzymatically oxidizing the enzyme substrate using an oxidizing agent to form the marker.3. The method of claim 2 , wherein enzymatically oxidizing the enzyme substrate using an oxidizing agent includes reducing the solubility or stability of the enzymatic substrate so that the enzymatic substrate becomes deposited as the marker.4. The method of claim 3 , wherein the enzymatic substrate is selected from the group consisting of a chromogen and a tyramide-conjugate.6. The method of claim 5 , wherein R claim 5 , R claim 5 , and Rare hydrogen and Ris hydroxyl.8. The method of claim 1 , wherein the enzyme is an oxidoreductase or a peroxidase.9. The method of claim 1 , wherein the enzyme is selected from horseradish peroxidase claim 1 , glutathione peroxidase claim 1 , and microoxidase.10. The method of claim 1 , wherein the specific binding moiety comprises an antibody or a nucleic acid.11. The method of claim 1 , wherein depositing the marker proximally to the target in the presence of the deposition enhancer includes the deposition enhancer at a concentration ranging from about 5 mM to about 15 mM.12. The method of claim 1 , wherein the enzyme substrate is selected from 1 claim 1 ,3-diaminobenzidine claim 1 , 3-amino-9-ethylcarbazole claim 1 , tetramethylbenzidine claim 1 , a fluorescein claim 1 , a luminophore claim 1 , a coumarin claim 1 , a BODIPY dye claim 1 , a resorufin claim 1 , a rhodamine claim 1 , or a derivative thereof.13. The method of claim 1 , wherein the enzyme substrate is a tyramine derivative.14. The method of claim 1 , wherein contacting the sample with a detection solution comprises exposing the sample to the ...

STABILIZED HEMATOXYLIN

A stabilized hematoxylin composition is disclosed that includes one or both of a host compound and an antioxidant. The disclosed composition exhibits sufficient stability to be utilized in an automated staining process without undue degradation prior to use of the composition to stain a biological sample. Methods of using and making the stabilized composition also are disclosed. 134.-. (canceled)35. A method for staining a biological sample , comprising:contacting the biological sample with a stabilized hematoxylin composition, the hematoxylin composition comprising a solvent, hematoxylin, an amount of a chemical oxidant sufficient to convert at least a portion of the hematoxylin to hematein, a mordant, an antioxidant, and a host compound, wherein the host compound is selected from the group consisting of a polysaccharide, a cryptand, a cryptophane, a cavitand, a crown ether, a dendrimer, a nanotube, a calixarene, a valinomycin, a nigericin, and a derivative thereof.36. The method of claim 35 , further comprising contacting the sample with a counterstain.37. The method of claim 36 , wherein the counterstain is selected from the group consisting of eosin Y claim 36 , orange G claim 36 , light green SF yellowish claim 36 , Bismark Brown claim 36 , fast green FCF claim 36 , OA-6 claim 36 , EA25 claim 36 , EA36 claim 36 , EA50 and EA65.38. The method of claim 35 , wherein contacting the sample with the hematoxylin composition comprises a progressive hematoxylin staining protocol.39. The method of claim 35 , wherein contacting the sample with the hematoxylin composition comprises a regressive hematoxylin staining protocol.40. The method of claim 35 , wherein the method is automated.41. The method of claim 35 , wherein the biological sample is supported on a substrate.42. The method of claim 41 , wherein the substrate comprises a microscope slide.43. The method of claim 35 , wherein the biological sample comprises a tissue section or a cytology sample.44. The method of ...

Antibody-Nanoparticle Conjugates and Methods for Making and Using Such Conjugates

Disclosed herein are antibody-nanoparticle conjugates that include two or more nanoparticles (such as gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof) directly linked to an antibody or fragment thereof through a metal-thiol bond. Methods of making the antibody-nanoparticle conjugates disclosed herein include reacting an arylphosphine-nanoparticle composite with a reduced antibody to produce an antibody-nanoparticle conjugate. Also disclosed herein are methods for detecting a target molecule in a sample that include using an antibody-nanoparticle conjugate (such as the antibody-nanoparticle conjugates described herein) and kits for detecting target molecules utilizing the methods disclosed herein. 1. An antibody-nanoparticle conjugate , comprising two or more nanoparticles directly linked to an antibody through a metal-thiol bond.2. The antibody-nanoparticle conjugate of claim 1 , wherein the conjugate comprises two to seven nanoparticles.3. The antibody-nanoparticle conjugate of claim 2 , wherein the conjugate comprises five nanoparticles.4. The antibody-nanoparticle conjugate of claim 1 , wherein the metal of the metal-thiol bond is conjugated to a cysteine residue of the antibody.5. (canceled)6. The antibody-nanoparticle conjugate of claim 1 , wherein the antibody comprises a rabbit claim 1 , goat claim 1 , mouse claim 1 , or anti-hapten antibody.7. The antibody-nanoparticle conjugate of claim 1 , wherein the nanoparticle comprises gold claim 1 , palladium claim 1 , platinum claim 1 , silver claim 1 , copper claim 1 , nickel claim 1 , cobalt claim 1 , iridium claim 1 , or an alloy of two or more thereof.8. The antibody-nanoparticle conjugate of claim 1 , wherein the nanoparticle is about 5 nm or less in diameter.9. The antibody-nanoparticle conjugate of claim 8 , wherein the nanoparticle is about 0.5 to 5 nm in diameter.1031.-. (canceled)32. A method for detecting a target molecule in a sample claim 8 , ...

HAPTEN CONJUGATES FOR TARGET DETECTION

Embodiments of hapten conjugates including a hapten, an optional linker, and a peroxidase-activatable aryl moiety are disclosed. In some embodiments, the peroxidase-activatable aryl moiety is tyramine or a tyramine derivative. Embodiments of methods for making and using the hapten conjugates also are disclosed. In particular embodiments, the hapten conjugates are used in a signal amplification assay. In certain embodiments, the hapten is an oxazole, a pyrazole, a thiazole, a benzofurazan, a triterpene, a urea, a thiourea other than a rhodamine thiourea, a nitroaryl other than dinitrophenyl or trinitrophenyl, a rotenoid, a cyclolignan, a heterobiaryl, an azoaryl, a benzodiazepine, or 7-diethylamino-3-carboxycoumarin. The hapten is coupled to the peroxidase-activatable aryl moiety directly or indirectly via a linker. In certain embodiments, the hapten conjugates are used in multiplexed assays. 1. A hapten conjugate , comprising:a hapten selected from an oxazole, a pyrazole, a thiazole, a benzofurazan, a triterpene, a urea, a thiourea other than a rhodamine thiourea, a nitroaryl other than dinitrophenyl or trinitrophenyl, a rotenoid, a cyclolignan, a heterobiaryl, an azoaryl, a benzodiazepine, 2,3,6,7-tetrahydro-11-oxo-1H,5H,11H-[1]benzopyrano[6,7,8-ij]quinolizine-10-carboxylic acid, or 7-diethylamino-3-carboxycoumarin;a linker; anda tyramine or a tyramine derivative.2. (canceled)6. (canceled)811.-. (canceled)1332.-. (canceled)33. A method , comprising:(a) immobilizing a first peroxidase on a first target in a sample, wherein the first peroxidase is capable of reacting with a peroxidase-activatable aryl moiety;(b) contacting the sample with a solution comprising a first hapten conjugate, the first hapten conjugate comprising a first hapten selected from an oxazole, a pyrazole, a thiazole, a benzofurazan, a triterpene, a urea, a thiourea other than a rhodamine thiourea, a nitroaryl other than dinitrophenyl or trinitrophenyl, a rotenoid, a cyclolignan, a heterobiaryl, an ...

POLYMER STABILIZATION OF CHROMOGEN SOLUTIONS

Disclosed embodiments concern a composition comprising DAB chromogen, and/or derivative thereof, a stabilizer, and polymer capable of preventing or reducing DAB precipitation relative to a composition that does not comprise the polymer. Also disclosed herein is a method for using the disclosed composition and embodiments of a kit. 2. The composition of claim 1 , wherein the polymer comprises a sulfate functional group claim 1 , a sulfonate functional group claim 1 , an amine functional groups claim 1 , or combinations thereof.3. The composition of claim 2 , wherein the amine functional groups is a primary amine claim 2 , a secondary amine claim 2 , a tertiary amine claim 2 , or a quaternary amine.4. The composition of claim 1 , wherein the polymer is a polyalkyleneamine polymer.5. The composition of claim 1 , wherein the polymer is selected from dextran sulfate claim 1 , polystyrene sulfonate claim 1 , polyethyleneimine claim 1 , aminodextran claim 1 , dextran DEAE claim 1 , polystyrene sulfonate maleic acid co-polymer claim 1 , polyvinylsulfonate claim 1 , poly(2-vinyl-1-methylpyridinium bromide claim 1 , poly(2-methylacryloxethyltrimethylammonium bromide claim 1 , poly(acrylamide/2-methylacryloxethyltrimethylammonium bromide claim 1 , and combinations thereof.6. The composition of claim 1 , wherein the polymer is a linear or branched polyethyleneimine.7. The composition of claim 6 , wherein the linear or branched polyethyleneimine is a 4 kDa HCl salt of polyethyleneimine.8. The composition of claim 1 , wherein the polymer has a number average molecular weight of between about 1 kDa and about 500 kDa.9. The composition of claim 1 , wherein the polymer is about 0.05 percent to about 10 percent of the solution by weight.10. The composition of claim 1 , wherein the DAB chromogen or the derivative thereof has a concentration of about 0.1 mM to about 100 mM.13. The composition of claim 1 , further comprising an enhancer selected from the group consisting of imidazole ...

METHOD FOR CHROMOGENIC DETECTION OF TWO OR MORE TARGET MOLECULES IN A SINGLE SAMPLE

The present invention provides a method and kit for detection of two or more target molecules in a single tissue sample, such as for gene and protein dual detection in a single tissue sample. Methods comprise treating a tissue sample with a first binding moiety that specifically binds a first target molecule. Methods further comprise treating the tissue sample with a solution containing a soluble electron-rich aromatic compound prior to or concomitantly with contacting the tissue sample with a hapten-labeled binding moiety and detecting a second target molecule. In one example, the first target molecule is a protein and the second is a nucleic acid sequence, the first target molecule being detected by immunohistochemistry and the second by in situ hybridization. The disclosed method reduces background due to non-specific binding of the hapten-labeled specific binding moiety to an insoluble electron rich compound deposited near the first target molecule. 3. The method of claim 2 , wherein the electron-rich aromatic compound comprises naphthol.4. The method of claim 3 , where the naphthol concentration is effective to allow dual nucleic acid and protein detection in a single sample and ranges from 1 milligrams per milliliter to 30 milligrams per milliliter.5. The method of claim 3 , where the naphthol concentration is effective to allow dual nucleic acid and protein detection in a single sample and ranges from 1 milligrams per milliliter to 7 milligrams per milliliter.6. The method of claim 3 , where the naphthol concentration ranges from about 0.3 milligrams per milliliter to about 1 milligram per milliliter.7. The method of claim 1 , where the hapten of the hapten-labeled nucleic acid probe is a nitroaryl compound.8. The method of claim 7 , where the nitroaryl compound is dinitrophenol.9. The method of claim 8 , where the concentration of the dinitrophenol nucleic acid-labeled probe is at least 5 μg/ml.10. The method of claim 9 , where the concentration of the ...

ANTIBODY-NANOPARTICLE CONJUGATES AND METHODS FOR MAKING AND USING SUCH CONJUGATES

Disclosed herein are antibody-nanoparticle conjugates that include two or more nanoparticles (such as gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof) directly linked to an antibody or fragment thereof through a metal-thiol bond. Methods of making the antibody-nanoparticle conjugates disclosed herein include reacting an arylphosphine-nanoparticle composite with a reduced antibody to produce an antibody-nanoparticle conjugate. Also disclosed herein are methods for detecting a target molecule in a sample that include using an antibody-nanoparticle conjugate (such as the antibody-nanoparticle conjugates described herein) and kits for detecting target molecules utilizing the methods disclosed herein. 131-. (canceled)32. A method for detecting a target molecule in a sample , comprising:(a) contacting a sample with a first antibody that binds to the target molecule;(b) contacting the sample with a second antibody conjugated to one or more enzyme molecules, wherein the second antibody specifically binds to the first antibody;(c) contacting the complex formed in step (b) above with an antibody-nanoparticle conjugate, that specifically binds to the second antibody, wherein said antibody-nanoparticle conjugate comprises at least two or more metal nanoparticles linked to a single antibody through two or more metal-thiol bonds, wherein the metal-thiol bonds are between thiol of cysteine residues of the antibody and the two or more metal nanoparticles without an intervening linker;(d) contacting the sample with substrate of the enzyme and a metal ion, such that a metal precipitate forms and colocalizes with the target molecule; and(e) detecting the metal precipitate, thereby detecting the target molecule.33. (canceled)34. (canceled)35. The method of claim 32 , wherein the target molecule comprises a hapten-labeled nucleic acid probe that specifically binds to the target molecule and wherein the first antibody specifically ...

MOLECULAR CONJUGATE

A method is disclosed for making a conjugate of two molecules using a hydrazide thiol linker. In a particular working embodiment, an Fc-specific antibody-enzyme conjugate is made using the method and demonstrated to provide exceptional staining sensitivity and specificity in immunohistochemical and in situ hybridization assays. 1. A kit , comprising: an antibody,', 'a detectable label comprising an enzyme including about 17-25 maleimide groups introduced into the enzyme by NHS-PEG-maleimide linkers, each linker being covalently bonded to an amino group of the enzyme, and', 'a plurality of hydrazide thiol linkers, each hydrazide thiol linker comprising one or more linking atoms positioned between a hydrazide group and a thiol group, wherein the hydrazide thiol linker is selected from mercaptobutyric acid hydrazide (MBH), mercaptobutyric acid carbohydrazide (MBCH), mercaptoacetamido-mercaptobutyric acid hydrazide (MAMBH), thiohexanamidomercaptobutyric acid hydrazide (THMBH), N,N′-(6-hydrazinyl-6-oxohexane-1,5-diyl)bis(2-mercaptoacetamide) (BTAL), bisthiohexanamidohydrazidolysine (BTHL), N-(1,5-dihydrazinyl-1,5-dioxopentan-2-yl)2-mercaptoacetamide (TAGD), thiohexamidoglutamic acid dihydrazide (THGD), a PEG-based hydrazide-thiol linker, a multifunctional hydrazide thiol linker, a PEG-based multifunctional hydrazide thiol linker, a polyacrylamide hydrazide thiol linker, and combinations thereof, the hydrazide group of each hydrazide thiol linker being covalently bonded to an oxidized glycosylated Fc portion of the antibody and the thiol group of one of the hydrazide thiol linkers being covalently bonded to the detectable label via one of the maleimide groups., 'a conjugate, comprising'}2. The kit of claim 1 , wherein the hydrazide thiol linker comprises a PEG-based hydrazide thiol linker.3. The kit of claim 2 , wherein the PEG-based hydrazide thiol linker comprises a mercapto-dPEG-hydrazide linker.4. The kit of claim 1 , wherein the hydrazide thiol linker comprises MBH ...

Passive, gravity-driven system for treatment of an effluent in a diagnostic system

Automated systems to make target compounds from slide stainer waste streams inactive utilizing advanced oxidation processes are described herein. Advanced oxidation processes are promoted by UV irradiation and further accelerated by use of radical initiators, such as hydrogen peroxide. The automated systems further include mechanisms for segregating components of the waste streams.

Antibody Conjugates

Antibody/signal-generating moiety conjugates are disclosed that include an antibody covalently linked to a signal-generating moiety through a heterobifunctional polyalkyleneglycol linker. The disclosed conjugates show exceptional signal-generation in immunohistochemical and in situ hybridization assays on tissue sections and cytology samples. In one embodiment, enzyme-metallographic detection of nucleic acid sequences with hapten-labeled probes can be accomplished using the disclosed conjugates as a primary antibody without amplification. 1. An antibody-signal-generating moiety conjugate comprising an antibody covalently linked to a signal-generating moiety through a heterobifunctional PEG linker.2. The conjugate of claim 1 , wherein a thiol-reactive group of the linker is covalently linked to the antibody and an amine-reactive group of the heterobifunctional linker is covalently linked to the signal-generating moiety.3. The conjugate of claim 2 , wherein the thiol-reactive group of the linker is covalently linked to a cysteine residue of the antibody.4. The conjugate of claim 2 , wherein the thiol-reactive group of the linker is covalently linked to a thiol group that is introduced to the antibody.5. The conjugate of claim 1 , wherein an aldehyde-reactive group of the linker is covalently linked to the antibody and a thiol-reactive group of the linker is covalently linked to the signal-generating moiety.6. The conjugate of claim 5 , wherein the aldehyde-reactive group of the linker is covalently linked to an aldehyde formed on a glycosylated portion of the antibody.9. The conjugate of claim 8 , wherein the signal-generating moiety comprises an enzyme.10. The conjugate of claim 9 , wherein the enzyme comprises horseradish peroxidase or alkaline phosphatase.11. The conjugate of claim 8 , wherein s.=2 to 6.12. The conjugate of claim 8 , wherein n=4 to 12.14. The conjugate of claim 13 , wherein the signal-generating moiety comprises an enzyme.15. The conjugate of claim ...

Molecular Conjugate

A method is disclosed for making a conjugate of two molecules using a hydrazide thiol linker. In a particular working embodiment, an Fc-specific antibody-enzyme conjugate is made using the method and demonstrated to provide exceptional staining sensitivity and specificity in immunohistochemical and in situ hybridization assays.

SIGNALING CONJUGATES AND METHODS OF USE

Disclosed herein are embodiments of a signaling conjugate, embodiments of a method of using the signaling conjugates, and embodiments of a kit comprising the signaling conjugate. The disclosed signaling conjugate comprises a latent reactive moiety and a chromogenic moiety that may further comprise a linker suitable for coupling the latent reactive moiety to the chromogenic moiety. The signaling conjugate may be used to detect one or more targets in a biological sample and are capable of being covalently deposited directly on or proximally to the target. Particular disclosed embodiments of the method of using the signaling conjugate comprise multiplexing methods. 128-. (canceled)29. A stained biological sample , comprising:(a) a binding complex comprising a target, a detection probe specifically bound to the target, and an enzyme conjugated to the detection probe, wherein the enzyme comprises a peroxidase, and wherein the target comprises a polypeptide, a nucleic acid, or a tyrosine-containing moiety;(b) a deposited complex comprising a signaling conjugate comprising a phenolic moiety and a chromogenic moiety, wherein the phenolic moiety of the signaling conjugate is covalently bound to: (1) a location on the binding complex proximate to or directly on the enzyme conjugate; (2) a location on the binding complex proximate to or directly on the target; or (3) a location on the biological sample proximate to the target;wherein the stained biological sample is immobilized on a solid support, and wherein the chromogenic moiety of the deposited complex produces a signal that provides for the specific detection of the target when exposed to light, wherein the light comprises one or more of visible light, infrared light, or near infrared light.30. A stained biological sample , comprising:(a) a binding complex comprising a target antigen, a detection probe specifically bound to the target antigen, and an enzyme conjugated to the detection probe, wherein the enzyme comprises a ...

Phenol-free acid-fast bacteria solution

The present disclosure relates to acid fast staining compositions which are free from phenol. The present disclosure also related to a method of detecting an acid fast organism in a biological sample comprising: (a) applying an acid fast staining composition to the biological sample, the acid fast staining solution comprising a fuchsin, a base, a surfactant, and an alcohol, and wherein the acid fast staining composition is free from phenol; and (b) incubating the biological sample with the acid fast staining composition for a predetermined amount of time at a predetermined temperature.

SIGNALING CONJUGATES AND METHODS OF USE

Disclosed herein are embodiments of a signaling conjugate, embodiments of a method of using the signaling conjugates, and embodiments of a kit comprising the signaling conjugate. The disclosed signaling conjugate comprises a latent reactive moiety and a chromogenic moiety that may further comprise a linker suitable for coupling the latent reactive moiety to the chromogenic moiety. The signaling conjugate may be used to detect one or more targets in a biological sample and are capable of being covalently deposited directly on or proximally to the target. Particular disclosed embodiments of the method of using the signaling conjugate comprise multiplexing methods. 128-. (canceled)30. The method of claim 29 , further comprising:(g) after step (f), analyzing the tissue sample by light microcopy such that, if the target antigen is present in the biological sample, the deposited chromogen produces a colored signal that provides for the detection of the target antigen when exposed to light, wherein the light comprises one or more of visible light, infrared light, or near infrared light.31. The method of claim 29 , wherein the target antigen comprises a polypeptide.32. The method of claim 29 , wherein the secondary antibody is an anti-species antibody against the species of the primary antibody claim 29 , wherein the primary antibody is a rabbit claim 29 , or mouse antibody and the secondary antibody is a goat antibody.33. The method of claim 29 , wherein the secondary antibody is an anti-hapten antibody.35. The method of claim 34 , further comprising:(g) after step (f), analyzing the tissue sample by light microcopy such that, if the target nucleic acid is present in the biological sample, the deposited chromogen produces a colored signal that provides for the detection of the target nucleic acid when exposed to light, wherein the light comprises one or more of visible light, infrared light, or near infrared light.36. The method of claim 34 , wherein the probe nucleic acid ...

Signaling conjugates and methods of use

Disclosed herein are embodiments of a signaling conjugate, embodiments of a method of using the signaling conjugates, and embodiments of a kit comprising the signaling conjugate. The disclosed signaling conjugate comprises a latent reactive moiety and a chromogenic moiety that may further comprise a linker suitable for coupling the latent reactive moiety to the chromogenic moiety. The signaling conjugate may be used to detect one or more targets in a biological sample and are capable of being covalently deposited directly on or proximally to the target. Particular disclosed embodiments of the method of using the signaling conjugate comprise multiplexing methods.

SIGNALING CONJUGATES AND METHODS OF USE

Disclosed herein are embodiments of a signaling conjugate, embodiments of a method of using the signaling conjugates, and embodiments of a kit comprising the signaling conjugate. The disclosed signaling conjugate comprises a latent reactive moiety and a chromogenic moiety that may further comprise a linker suitable for coupling the latent reactive moiety to the chromogenic moiety. The signaling conjugate may be used to detect one or more targets in a biological sample and are capable of being covalently deposited directly on or proximally to the target. Particular disclosed embodiments of the method of using the signaling conjugate comprise multiplexing methods. 128-. (canceled)31. The method of claim 29 , wherein the target antigen comprises a polypeptide.32. The method of claim 30 , wherein the target antigen comprises a polypeptide.33. The method of claim 29 , wherein the secondary antibody is an anti-species antibody against the species of the primary antibody claim 29 , wherein the primary antibody is a rabbit claim 29 , or mouse antibody and the secondary antibody is a goat antibody.34. The method of claim 30 , wherein the secondary antibody is an anti-species antibody against the species of the primary antibody claim 30 , wherein the primary antibody is a rabbit claim 30 , or mouse antibody and the secondary antibody is a goat antibody.35. The method of claim 29 , wherein the secondary antibody is an anti-hapten antibody.36. The method of claim 30 , wherein the secondary antibody is an anti-hapten antibody.37. The method of claim 29 , further comprising claim 29 , prior to step (a) claim 29 , inactivating endogenous peroxidase activity in said sample with a peroxidase inhibitor.38. The method of claim 30 , further comprising claim 30 , prior to step (a) claim 30 , inactivating endogenous peroxidase activity in said sample with a peroxidase inhibitor.39. The method according to claim 29 , wherein the signaling conjugate further comprises a linker joining the ...

ANTIBODY-NANOPARTICLE CONJUGATES AND METHODS FOR MAKING AND USING SUCH CONJUGATES

Disclosed herein are antibody-nanoparticle conjugates that include two or more nanoparticles (such as gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof) directly linked to an antibody or fragment thereof through a metal-thiol bond. Methods of making the antibody-nanoparticle conjugates disclosed herein include reacting an arylphosphine-nanoparticle composite with a reduced antibody to produce an antibody-nanoparticle conjugate. Also disclosed herein are methods for detecting a target molecule in a sample that include using an antibody-nanoparticle conjugate (such as the antibody-nanoparticle conjugates described herein) and kits for detecting target molecules utilizing the methods disclosed herein. 19-. (canceled)10. A method for producing an antibody-nanoparticle conjugate , comprising reacting an arylphosphine-nanoparticle composite with a reduced antibody , thereby producing the antibody-nanoparticle conjugate.11. The method of claim 10 , further comprising forming a reduced antibody.12. The method of claim 11 , wherein the reduced antibody is formed by reacting an antibody with a reducing agent to produce the reduced antibody.13. The method of claim 12 , wherein the reducing agent is a mono-thiol compound or a di-thiol compound.14. The method of claim 13 , wherein the reducing agent is dithiothreitol.15. The method of claim 10 , further comprising adjusting reactant stoichiometry and reaction duration to couple two or more nanoparticles to the reduced antibody.16. The method of claim 15 , wherein the antibody-nanoparticle conjugate comprises two to seven nanoparticles.17. The method of claim 16 , wherein the antibody-nanoparticle conjugate comprises five nanoparticles.18. The method of claim 15 , wherein the reactant stoichiometry of the arylphosphine-nanoparticle composite to the reduced antibody is about 7:1 claim 15 , and wherein the number of nanoparticles coupled to the reduced antibody is about five.19. ...

Biological sample processing composition and method

A method and composition are disclosed that are useful for processing biological samples. In one aspect, a biological sample such as a tissue section is treated using a histochemical technique and is contacted with a lipid compound during the process to enhance the definition of cellular and sub-cellular features that are observable in the sample when it is viewed microscopically. In other aspects, a coverslipping composition that includes a lipid compound and a method of coverslipping a sample using the coverslipping composition are disclosed.

Biological sample processing composition and method

A method and composition are disclosed that are useful for processing biological samples. In one aspect, a biological sample such as a tissue section is treated using a histochemical technique and is contacted with a lipid compound during the process to enhance the definition of cellular and sub-cellular features that are observable in the sample when it is viewed microscopically. In other aspects, a coverslipping composition that includes a lipid compound and a method of coverslipping a sample using the coverslipping composition are disclosed.

Biological sample processing composition and method

A method and composition are disclosed that are useful for processing biological samples. In one aspect, a biological sample such as a tissue section is treated using a histochemical technique and is contacted with a lipid compound during the process to enhance the definition of cellular and sub-cellular features that are observable in the sample when it is viewed microscopically. In other aspects, a coverslipping composition that includes a lipid compound and a method of coverslipping a sample using the coverslipping composition are disclosed.

Biological sample processing composition and method

The present invention relates to a method of processing a biological sample on a microscope slide.

Biological sample processing composition and method

A method and composition are disclosed that are useful for processing biological samples. In one aspect, a biological sample such as a tissue section is treated using a histochemical technique and is contacted with a lipid compound during the process to enhance the definition of cellular and sub-cellular features that are observable in the sample when it is viewed microscopically. In other aspects, a coverslipping composition that includes a lipid compound and a method of coverslipping a sample using the coverslipping composition are disclosed.

Biological sample processing composition and method

The present invention relates to a method of processing a biological sample on a microscope slide.

Biological sample processing composition and method

A method and composition are disclosed that are useful for processing biological samples. In one aspect, a biological sample such as a tissue section is treated using a histochemical technique and is contacted with a lipid compound during the process to enhance the definition of cellular and sub-cellular features that are observable in the sample when it is viewed microscopically. In other aspects, a coverslipping composition that includes a lipid compound and a method of coverslipping a sample using the coverslipping composition are disclosed.

Biological sample processing composition and method

A method and composition are disclosed that are useful for processing biological samples. In one aspect, a biological sample such as a tissue section is treated using a histochemical technique and is contacted with a lipid compound during the process to enhance the definition of cellular and sub-cellular features that are observable in the sample when it is viewed microscopically. In other aspects, a coverslipping composition that includes a lipid compound and a method of coverslipping a sample using the coverslipping composition are disclosed.

Biological sample processing composition and method

Stabilized hematoxylin

A stabilized hematoxylin composition is disclosed that includes one or both of a host compound and an antioxidant. The disclosed composition exhibits sufficient stability to be utilized in an automated staining process without undue degradation prior to use of the composition to stain a biological sample. Methods of using and making the stabilized composition also are disclosed.

Biological sample processing composition and method

Abstract A method and composition are disclosed that are useful for processing biological samples. In one aspect, a biological sample such as a tissue section is treated using a histochemical technique and is contacted with a lipid compound during the process to enhance the definition of cellular and sub-cellular features that are observable in the sample when it is viewed microscopically. In other aspects, a coverslipping composition that includes a lipid compound and a method of coverslipping a sample using the coverslipping composition are disclosed. WO 2007/084429 PCT/US2007/000980 Move to De-paraffinizer/Stainer/ Solvent Exchanger De-paraffinize with Limonene Alcohol Rinse Slide Prep/Water Dispense Hematoxylin/Bluing Slide Prep/Water/50% alcohol Dispense Eosin Differentiation with 50% Alcohol Dehydration with 100% Alcohol Move to Convection Oven Heat at 45*C for 5.5 Minutes Move to Coverslipper -Dispense 20% Lipid Compound in Limonene/Add Coverslip Move to Convection Oven Cure at 70 0C for 5.5 Minutes

Signaling conjugates and methods of use

Disclosed herein are embodiments of a signaling conjugate, embodiments of a method of using the signaling conjugates, and embodiments of a kit comprising the signaling conjugate. The disclosed signaling conjugate comprises a latent reactive moiety and a chromogenic moiety that may further comprise a linker suitable for coupling the latent reactive moiety to the chromogenic moiety. The signaling conjugate may be used to detect one or more targets in a biological sample and are capable of being covalently deposited directly on or proximally to the target. Particular disclosed embodiments of the method of using the signaling conjugate comprise multiplexing methods.

Polymer stabilization of chromogen solutions

Disclosed embodiments concern a composition comprising DAB chromogen, and/or derivative thereof, a stabilizer, and polymer capable of preventing or reducing DAB precipitation relative to a composition that does not comprise the polymer. Also disclosed herein is a method for using the disclosed composition and embodiments of a kit.

Enzyme-catalyzed metal deposition for the enhanced in situ detection of immunohistochemical epitopes and nucleic acid sequences

The invention is directed to novel compositions of matter and methods of detecting in situ an immunohistochemical epitope or nucleic acid sequence of interest in a biological sample comprising binding an enzyme-labeled conjugate molecule to the epitope or sequence of interest in the presence of a redox-inactive reductive species and a soluble metal ion, thereby facilitating the reduction of the metal ion to a metal atom at or about the point where the enzyme is anchored. Novel phosphate derivatives of reducing agents are described that when exposed to a phosphatase are activated to their reducing form, thereby reducing metal ions to insoluble metal.

Phenol-free acid-fast staining composition and use thereof

The present disclosure relates to acid fast staining compositions which are free from phenol. The present disclosure also related to a method of detecting an acid fast organism in a biological sample comprising: (a) applying an acid fast staining composition to the biological sample, the acid fast staining solution comprising a fuchsin, a base, a surfactant, and an alcohol, and wherein the acid fast staining composition is free from phenol; and (b) incubating the biological sample with the acid fast staining composition for a predetermined amount of time at a predetermined temperature.

Haptens, hapten conjugates, compositions thereof and method for their preparation and use

A method for performing a multiplexed diagnostic assay, such as for two or more different targets in a sample, is described. One embodiment comprised contacting the sample with two or more specific binding moieties that bind specifically to two or more different targets. The two or more specific binding moieties are conjugated to different haptens, and at least one of the haptens is an oxazole, a pyrazole, a thiazole, a nitroaryl compound other than dinitrophenyl, a benzofurazan, a triterpene, a urea, a thiourea, a rotenoid, a coumarin, a cyclolignan, a heterobiaryl, an azo aryl, or a benzodiazepine. The sample is contacted with two or more different anti-hapten antibodies that can be detected separately. The two or more different anti-hapten antibodies may be conjugated to different detectable labels.

Stabilized hematoxylin

Signaling conjugates and methods of use

Disclosed herein are embodiments of a signaling conjugate, embodiments of a method of using the signaling conjugates, and embodiments of a kit comprising the signaling conjugate. The disclosed signaling conjugate comprises a latent reactive moiety and a chromogenic moiety that may further comprise a linker suitable for coupling the latent reactive moiety to the chromogenic moiety. The signaling conjugate may be used to detect one or more targets in a biological sample and are capable of being covalently deposited directly on or proximally to the target. Particular disclosed embodiments of the method of using the signaling conjugate comprise multiplexing methods.

Semi-synthesis and use of racemic hematoxylin

A racemic hematoxylin formulation is disclosed that includes one or both of a stabilizer compound and an antioxidant. The disclosed composition exhibits sufficient stability to be utilized in an automated staining process. Methods of using and making the stabilized composition also are disclosed.

Antibody conjugates via heterobifunctional peg linkers

Antibody/signal-generating moiety conjugates are disclosed that include an antibody covalently linked to a signal-generating moiety through a heterobifunctional polyalkyleneglycol linker. The disclosed conjugates show exceptional signal-generation in immunohistochemical and in situ hybridization assays on tissue sections and cytology samples. In one embodiment, enzyme-metallographic detection of nucleic acid sequences with hapten-labeled probes can be accomplished using the disclosed conjugates as a primary antibody without amplification.

Stabilized hematoxylin

Molecular conjugate

A method is disclosed for making a conjugate of two molecules using a hydrazide thiol linker. In a particular working embodiment, an Fc-specifÊc antibody- enzyme conjugate is made using the method and demonstrated to provide exceptional staining sensitivity and specificity in immunohistochemical and in situ hybridization assays.

Enzyme-catalyzed metal deposition for the enhanced in situ detection of immunohistochemical epitopes and nucleic acid sequences

The invention is directed to novel compositions of matter and methods of detecting in situ an immunohistochemical epitope or nucleic acid sequence of interest in a biological sample comprising binding an enzyme-labeled conjugate molecule to the epitope or sequence of interest in the presence of a redox-inactive reductive species and a soluble metal ion, thereby facilitating the reduction of the metal ion to a metal atom at or about the point where the enzyme is anchored. Novel phosphate derivatives of reducing agents are described that when exposed to a phosphatase are activated to their reducing form, thereby reducing metal ions to insoluble metal.

Signaling conjugates and methods of use

Disclosed herein are embodiments of a signaling conjugate, embodiments of a method of using the signaling conjugates, and embodiments of a kit comprising the signaling conjugate. The disclosed signaling conjugate comprises a latent reactive moiety and a chromogenic moiety that may further comprise a linker suitable for coupling the latent reactive moiety to the chromogenic moiety. The signaling conjugate may be used to detect one or more targets in a biological sample and are capable of being covalently deposited directly on or proximally to the target. Particular disclosed embodiments of the method of using the signaling conjugate comprise multiplexing methods.

Passive, gravity-driven system for treatment of an effluent in a diagnostic system

Passive, gravity-driven system for treatment of an effluent in a diagnostic system

Automated systems to make target compounds from slide stainer waste streams inactive utilizing advanced oxidation processes are described herein. Advanced oxidation processes are promoted by UV irradiation and further accelerated by use of radical initiators, such as hydrogen peroxide. The automated systems further include mechanisms for segregating components of the waste streams.

method to detect a target in a sample and kit

método para detectar um alvo em uma amostra, composição e kit [001]a presente invenção refere-se a composições que intensificam a deposição de porções detectáveis em amostras de tecido, os métodos utilizam essas composições e kits que incluem essas composições. as composições incluem um intensificador de deposição que tem uma fórmula e/ou em que r1, r2, r3, e r4 são independentemente selecionados a partir de alifático, arila, halogênio, uma porção que contém heteroátomo, e hidrogênio; r1 e/ou r3 podem ser iigados a r2 para formar um sistema de anel aromático, fundido; r5 é selecionado a partir de uma porção que contém heteroátomo; a é selecionado a partir de um átomo de carbono, um heteroátomo, diferente de enxofre, e qualquer combinação dos mesmos; n é 1 a 5, uma enzima, uma porção de ligação específica e uma porção detectável. method for detecting a target in a sample, composition and kit [001] the present invention relates to compositions that enhance the deposition of detectable portions in tissue samples, the methods use those compositions and kits that include those compositions. the compositions include a deposition enhancer that has a formula and / or in which r1, r2, r3, and r4 are independently selected from aliphatic, aryl, halogen, a portion containing heteroatom, and hydrogen; r1 and / or r3 can be linked to r2 to form a fused aromatic ring system; r5 is selected from a portion that contains heteroatom; a is selected from a carbon atom, a heteroatom, other than sulfur, and any combination thereof; n is 1 to 5, an enzyme, a specific binding portion and a detectable portion.

Hapten conjugates for target detection

Embodiments of hapten conjugates including a hapten, an optional linker, and a peroxidase- activatable aryl moiety are disclosed. In some embodiments, the peroxidase- activatable aryl moiety is tyramine or a tyramine derivative. Embodiments of methods for making and using the hapten conjugates also are disclosed. In particular embodiments, the hapten conjugates are used in a signal amplification assay. In certain embodiments, the hapten is an oxazole, a pyrazole, a thiazole, a benzofurazan, a triterpene, a urea, a thiourea other than a rhodamine thiourea, a nitroaryl other than dinitrophenyl or trinitrophenyl, a rotenoid, a cyclolignan, a heterobiaryl, an azoaryl, a benzodiazepine, or 7-diethylamino-3- carboxycoumarin. The hapten is coupled to the peroxidase-activatable aryl moiety directly or indirectly via a linker. In certain embodiments, the hapten conjugates are used in multiplexed assays.

METAL DEPOSIT CATALYZED BY AN ENZYME FOR IMPROVED IN SITU DETECTION OF IMMUNOHISTOCHEMICAL EPITHOPES AND NUCLEIC ACID SEQUENCES.

Un método de detección in situ de un epítopo inmunohistoquímico o una secuencia de ácido nucleico de interés en una muestra biológica, que comprende unir una molécula conjugada marcada con una enzima al epítopo o secuencia de interés en presencia de una especie reductora sin actividad redox y un ión metálico soluble, en el que dicha especie reductora sin actividad redox es un sustrato para dicha molécula conjugada marcada con una enzima y en el que dicha enzima de dicha molécula conjugada marcada con una enzima cataliza la transformación de la especie reductora sin actividad redox en un agente reductor, reduciendo de este modo el ión metálico a partículas metálicas insolubles en estado de oxidación 0, en o alrededor del punto en el que se ancla la enzima. A method of in situ detection of an immunohistochemical epitope or a nucleic acid sequence of interest in a biological sample, comprising binding a conjugated molecule labeled with an enzyme to the epitope or sequence of interest in the presence of a reducing species without redox activity and a soluble metal ion, wherein said reducing species without redox activity is a substrate for said conjugate molecule labeled with an enzyme and wherein said enzyme of said conjugated molecule labeled with an enzyme catalyzes the transformation of the reducing species without redox activity into a reducing agent, thereby reducing the metal ion to insoluble metal particles in oxidation state 0, at or around the point where the enzyme is anchored.

Microwave mediated synthesis of nucleic acid probes

A method for preparing a nucleic acid probe is provided. The method comprises forming an activated cytosine or cytidine by a bisulfite catalyzed reaction; and covalently linking a reporter molecule to the activated cytosine or cytidine, wherein said activating step, said covalently linking step, or both are conducted in the presence of microwave energy. Also provided by the invention are nucleic acid probes.

Polymer stabilization of chromogen solutions

Disclosed embodiments concern a composition comprising DAB chromogen, and/or derivative thereof, a stabilizer, and polymer capable of preventing or reducing DAB precipitation relative to a composition that does not comprise the polymer. Also disclosed herein is a method for using the disclosed composition and embodiments of a kit.

Antibody conjugates

Antibody/signal-generating moiety conjugates are disclosed that include an antibody covalently linked to a signal-generating moiety through a heterobifunctional polyalkyleneglycol linker. The disclosed conjugates show exceptional signal-generation in immunohistochemical and in situ hybridization assays on tissue sections and cytology samples. In one embodiment, enzyme-metallographic detection of nucleic acid sequences with hapten-labeled probes can be accomplished using the disclosed conjugates as a primary antibody without amplification.

Haptens, hapten conjugates, compositions thereof and method for their preparation and use

A method for performing a multiplexed diagnostic assay, such as for two or more different targets in a sample, is described. One embodiment comprised contacting the sample with two or more specific binding moieties that bind specifically to two or more different targets. The two or more specific binding moieties are conjugated to different haptens, and at least one of the haptens is an oxazole, a pyrazole, a thiazole, a nitroaryl compound other than dinitrophenyl, a benzofurazan, a triterpene, a urea, a thiourea, a rotenoid, a coumarin, a cyclolignan, a heterobiaryl, an azo aryl, or a benzodiazepine. The sample is contacted with two or more different anti-hapten antibodies that can be detected separately. The two or more different anti-hapten antibodies may be conjugated to different detectable labels.

Enhanced deposition of chromogens utilizing pyrimidine analogs

This disclosure relates to compositions that enhance the deposition of detectable moieties on tissue samples, methods utilizing these compositions and kits including these compositions. The compositions include a deposition enhancer having a formula and/or where R1, R2, R3, and R4 are independently selected from aliphatic, aryl, halogen, a heteroatom-containing moiety, and hydrogen; R1 and/or R3 can be bound to R2 to form a fused, aromatic ring system; R5 is selected from a heteroatom-containing moiety; A is selected from, a carbon atom, a heteroatom, other than sulfur, and any combination thereof; n is 1-5, an enzyme, a specific binding moiety and a detectable moiety.

Antibody-nanoparticle conjugates and methods for making and using such conjugates

Disclosed herein are antibody-nanoparticle conjugates that include two or more nanoparticles (such as gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof) directly linked to an antibody or fragment thereof through a metal-thiol bond. Methods of making the antibody- nanoparticle conjugates disclosed herein include reacting an arylphosphine-nanoparticle composite with a reduced antibody to produce an antibody-nanoparticle conjugate. Also disclosed herein are methods for detecting a target molecule in a sample that include using an antibody-nanoparticle conjugate (such as the antibody- nanoparticle conjugates described herein) and kits for detecting target molecules utilizing the methods disclosed herein.

A method for detecting a target in a sample by proximally depositing a marker, a composition for detecting a target in a sample by proximally depositing a marker and a kit comprising a detection solution.

método para detectar um alvo em uma amostra mediante deposição de modo proximal de um marcador, composição para detectar um alvo em uma amostra mediante deposição de modo proximal de um marcador e kit que compreende uma solução de detecção esta descrição refere-se a composições que realçam a deposição de porções químicas detectáveis em amostras de tecido, sendo que os métodos utilizam essas composições e kits que incluem essas composições. as composições incluem um realçador de deposição que tem uma fórmula e/ou em que r^ 1^, r^ 2^, r^ 3^ e r^ 4^ são independetemente selecionados dentre uma porção química que contém heteroátomo, alifática, arila, halogênio e hidrogênio; r^ 1^ e/ou r^ 3^ podem ser ligados a r^ 2^ para formar um sistema de anel aromático fundido; r^ 5^ é selecionado dentre uma porção química que contém heteroátomo; a é selecionado dentre um átomo de carbono, um heteroátomo, exceto enxofre, e qualquer combinação dos mesmos; n é 1 a 5, uma enzima, uma porção química de ligação específica e uma porção química detectável. A method for detecting a target in a sample by proximally depositing a marker, A composition for detecting a target in a sample by proximally depositing a marker and a kit comprising a detection solution This description relates to compositions which enhance deposition of detectable chemical portions in tissue samples, the methods using such compositions and kits comprising such compositions. compositions include a deposition enhancer having a formula and / or wherein r ^ 1 ^, r ^ 2 ^, r ^ 3 ^ and r ^ 4 ^ are independently selected from a chemical moiety containing heteroatom, aliphatic, aryl, halogen and hydrogen; R2 and / or R2 may be attached to R2 to form a fused aromatic ring system; R 5 is selected from a chemical moiety containing heteroatom; a is selected from a carbon atom, a heteroatom, except sulfur, and any combination thereof; n is 1 to 5, an enzyme, a specific binding chemical moiety and a detectable chemical moiety.

Antibody-nanoparticle conjugates and methods for making and using such conjugates

Disclosed herein are antibody-nanoparticle conjugates that include two or more nanoparticles (such as gold, palladium, platinum, silver, copper, nickel, cobalt, iridium, or an alloy of two or more thereof) directly linked to an antibody or fragment thereof through a metal-thiol bond. Methods of making the antibody- nanoparticle conjugates disclosed herein include reacting an arylphosphine-nanoparticle composite with a reduced antibody to produce an antibody-nanoparticle conjugate. Also disclosed herein are methods for detecting a target molecule in a sample that include using an antibody-nanoparticle conjugate (such as the antibody- nanoparticle conjugates described herein) and kits for detecting target molecules utilizing the methods disclosed herein.

Enhanced deposition of chromogens utilizing pyrimidine analogs

This disclosure relates to compositions that enhance the deposition of detectable moieties on tissue samples, methods utilizing these compositions and kits including these compositions. The compositions include a deposition enhancer having a formula and/or where R1, R2, R3, and R4 are independently selected from aliphatic, aryl, halogen, a heteroatom-containing moiety, and hydrogen; R1 and/or R3 can be bound to R2 to form a fused, aromatic ring system; R5 is selected from a heteroatom-containing moiety; A is selected from, a carbon atom, a heteroatom, other than sulfur, and any combination thereof; n is 1-5, an enzyme, a specific binding moiety and a detectable moiety.

Antibody-enzyme conjugate

Stabilization of polymers of chromogen solutions

Una composición para inmunohistoquímica cromogénica que comprende en una solución: a) una solución acuosa tamponada, b) un cromógeno de diaminobencidina (DAB), c) un estabilizador, y d) un polímero de polietilenimina soluble en la solución acuosa tamponada, en la que la precipitación del cromógeno DAB de la solución se reduce en condiciones de envejecimiento acelerado con respecto a una composición similar que no comprende el polímero de polietilenimina, en la que el estabilizador es metabisulfito sódico o bisulfito sódico. A composition for chromogenic immunohistochemistry comprising in a solution: a) a buffered aqueous solution, b) a diaminobenzidine (DAB) chromogen, c) a stabilizer, and d) a polyethyleneimine polymer soluble in the buffered aqueous solution, in which the precipitation of DAB chromogen from solution is reduced under accelerated aging conditions relative to a similar composition that does not comprise the polyethyleneimine polymer, in which the stabilizer is sodium metabisulfite or sodium bisulfite.

Method of preparing and use of prodrugs of betulinic acid derivatives

A composition and method of preventing or inhibiting tumor growth and, more particularly, of treating a malignant tumor, using prodrugs of plant-derived compounds and derivatives is disclosed. In the method, a composition containing betulinic acid or a betulinic acid derivative is administered in a prodrug form to release betulinic acid or a betulinic acid derivative in vivo at the tumor site.

Enhanced deposition of chromogens utilizing pyrimidine analogs

This disclosure relates to compositions that enhance the deposition of detectable moieties on tissue samples, methods utilizing these compositions and kits including these compositions. The compositions include a deposition enhancer having a formula and/or where R1, R2, R3, and R4 are independently selected from aliphatic, aryl, halogen, a heteroatom-containing moiety, and hydrogen; R1 and/or R3 can be bound to R2 to form a fused, aromatic ring system; R5 is selected from a heteroatom-containing moiety; A is selected from, a carbon atom, a heteroatom, other than sulfur, and any combination thereof; n is 1-5, an enzyme, a specific binding moiety and a detectable moiety.

Molecular conjugate

The present invention relates to conjugates comprising an antibody covalently bonded to a detectable label through a hydrazide thiol linker, wherein a hydrazide group of the linker is covalently bonded to an Fc region of the antibody, and wherein the detectable label is an enzyme having at least one thiol reactive group, wherein a thiol group of the hydrazide thiol linker is coupled to the at least one thiol reactive group of the enzyme. The present invention further relates to conjugates of an antibody and a modified phosphatase enzyme, the modified phosphatase enzyme comprising at least one thiol reactive group, wherein the antibody and the modified phosphatase enzyme are coupled through a mercaptobutyric acid hydrazide linker, and wherein a hydrazide group of the mercaptobutyric acid hydrazide linker is covalently bonded to an Fc region of the antibody.

Hapten conjugates for target detection

Embodiments of hapten conjugates including a hapten, an optional linker, and a peroxidase- activatable aryl moiety are disclosed. In some embodiments, the peroxidase- activatable aryl moiety is tyramine or a tyramine derivative. Embodiments of methods for making and using the hapten conjugates also are disclosed. In particular embodiments, the hapten conjugates are used in a signal amplification assay. In certain embodiments, the hapten is an oxazole, a pyrazole, a thiazole, a benzofurazan, a triterpene, a urea, a thiourea other than a rhodamine thiourea, a nitroaryl other than dinitrophenyl or trinitrophenyl, a rotenoid, a cyclolignan, a heterobiaryl, an azoaryl, a benzodiazepine, or 7-diethylamino-3- carboxycoumarin. The hapten is coupled to the peroxidase-activatable aryl moiety directly or indirectly via a linker. In certain embodiments, the hapten conjugates are used in multiplexed assays.

Enhanced deposition of chromogens utilizing pyrimidine analogs

Signaling conjugates and methods of use

Disclosed herein are embodiments of a signaling conjugate, embodiments of a method of using the signaling conjugates, and embodiments of a kit comprising the signaling conjugate. The disclosed signaling conjugate comprises a latent reactive moiety and a chromogenic moiety that may further comprise a linker suitable for coupling the latent reactive moiety to the chromogenic moiety. The signaling conjugate may be used to detect one or more targets in a biological sample and are capable of being covalently deposited directly on or proximally to the target. Particular disclosed embodiments of the method of using the signaling conjugate comprise multiplexing methods.

Passive, gravity-driven system for treatment of an effluent in a diagnostic system

Automated systems to make target compounds from slide stainer waste streams inactive utilizing advanced oxidation processes are described herein. Advanced oxidation processes are promoted by UV irradiation and further accelerated by use of radical initiators, such as hydrogen peroxide. The automated systems further include mechanisms for segregating components of the waste streams.

Enzymkatalysierte metalldeposition zum verstärkten in-situ-nachweis von immunhistologischen epitopen und nukleinsäuresequenzen

Methods and systems for preparing and analyzing cellular samples for morphological characteristics and biomarker expression

This disclosure relates generally to the use of automated platforms in the preparation of biomarker-stained cellular samples for microscopic analysis and use of such stained cells in the diagnosis of certain conditions. Disclosed herein is a method of affinity staining a Romanowsky-type stained sample on automated advanced staining systems, wherein the automated advanced stainer destains the sample prior to contact with a biomarker-specific reagent. Also disclosed herein are methods of processing body fluid samples for morphological and biomarker analysis by depositing cells of the sample in a thin layer onto one or more solid supports, staining at least one such solid support with a Romanowsky-type stain and staining at least one such solid support for one or more biomarkers useful for categorizing one or more cells of the sample.

Digitally enhanced microscopy for multiplexed histology

Disclosed herein are embodiments of imaging biological specimens. An imaging system can include a microscope for directly viewing the biological specimen and a multi-spectral imaging apparatus for outputting digitally enhanced images, near-video rate imaging, and/or videos of the specimen. An imaging system can include a digital scanner that digitally processes images to produce a composite image with enhanced color contrast of features of interest.

Polymer stabilization of chromogen solutions

Disclosed embodiments concern a composition comprising DAB chromogen, and/or derivative thereof, a stabilizer, and polymer capable of preventing or reducing DAB precipitation relative to a composition that does not comprise the polymer. Also disclosed herein is a method for using the disclosed composition and embodiments of a kit.

Haptens, hapten conjugates, compositions thereof preparation and method for their preparation and use

A method for performing a multiplexed diagnostic assay, such as for two or more different targets in a sample, is described. One embodiment comprised contacting the sample with two or more specific binding moieties that bind specifically to two or more different targets. The two or more specific binding moieties are conjugated to different haptens, and at least one of the haptens is an oxazole, a pyrazole, a thiazole, a nitroaryl compound other than dinitrophenyl, a benzofurazan, a triterpene, a urea, a thiourea, a rotenoid, a coumarin, a cyclolignan, a heterobiaryl, an azo aryl, or a benzodiazepine. The sample is contacted with two or more different anti-hapten antibodies that can be detected separately. The two or more different anti-hapten antibodies may be conjugated to different detectable labels.

Enhanced deposition of chromogens utilizing pyrimidine analogs

This disclosure relates to compositions that enhance the deposition of detectable moieties on tissue samples, methods utilizing these compositions and kits including these compositions. The compositions include a deposition enhancer having a formula where R 1 , R 2 , R 3 , and R 4 are independently selected from aliphatic, aryl, halogen, a heteroatom-containing moiety, and hydrogen; R 1 and/or R 3 can be bound to R 2 to form a fused, aromatic ring system; R 5 is selected from a heteroatom-containing moiety; A is selected from, a carbon atom, a heteroatom, other than sulfur, and any combination thereof; n is 1-5, an enzyme, a specific binding moiety and a detectable moiety.