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WO1996000902A1 - Colorants a base de cyanine substitues par des ions n-heteroaromatiques et par des ions iminium, utilises comme marques fluorescentes - Google Patents

Colorants a base de cyanine substitues par des ions n-heteroaromatiques et par des ions iminium, utilises comme marques fluorescentes Download PDF

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Publication number
WO1996000902A1
WO1996000902A1 PCT/US1995/008778 US9508778W WO9600902A1 WO 1996000902 A1 WO1996000902 A1 WO 1996000902A1 US 9508778 W US9508778 W US 9508778W WO 9600902 A1 WO9600902 A1 WO 9600902A1
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WO
WIPO (PCT)
Prior art keywords
group
taken together
cyanine dye
ring
aromatic ring
Prior art date
Application number
PCT/US1995/008778
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English (en)
Inventor
Linda G. Lee
Sam L. Woo
Original Assignee
Biometric Imaging, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US08/268,852 external-priority patent/US5453505A/en
Application filed by Biometric Imaging, Inc. filed Critical Biometric Imaging, Inc.
Priority to EP95926272A priority Critical patent/EP0769145A1/fr
Priority to AU30085/95A priority patent/AU3008595A/en
Publication of WO1996000902A1 publication Critical patent/WO1996000902A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
    • A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0058—Antibodies
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B23/00—Methine or polymethine dyes, e.g. cyanine dyes
    • C09B23/0008—Methine or polymethine dyes, e.g. cyanine dyes substituted on the polymethine chain
    • C09B23/0041—Methine or polymethine dyes, e.g. cyanine dyes substituted on the polymethine chain the substituent being bound through a nitrogen atom
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B23/00—Methine or polymethine dyes, e.g. cyanine dyes
    • C09B23/0066—Methine or polymethine dyes, e.g. cyanine dyes the polymethine chain being part of a carbocyclic ring,(e.g. benzene, naphtalene, cyclohexene, cyclobutenene-quadratic acid)
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B23/00—Methine or polymethine dyes, e.g. cyanine dyes
    • C09B23/02—Methine or polymethine dyes, e.g. cyanine dyes the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/533—Production of labelled immunochemicals with fluorescent label

Definitions

  • the present invention relates to cyanine dyes for use as fluorescent probes. More specifically, the present invention relates to cyanine dyes substituted with either an N-heteroaromatic ion or an iminium ion, the ion reducing the aggregation of the cyanine dyes and enhancing the photostability of the dyes.
  • Fluorescent dyes have a wide variety of uses including the labeling of antibodies, D ⁇ A, carbohydrates and cells. In order for a fluorescent dye to function as a label, the dye must bind to the molecule or cell to be labeled. Fluorescent labels are therefore designed to include at least one reactive moiety which reacts with amino, hydroxy and/or sulfhydryl nucleophiles present on the molecules being labeled.
  • Suitable reactive moieties include carboxylic acids, acid halides, sulfonic acids, esters, aldehydes, disulfides, isothiocyanates, isocyanates, monochlorotriazine, dichlorotriazine, mono- or di- halogen substituted pyridines, mono- or di-halogen substituted diazines, maleimide, aziridines, sulfonyl halides, hydroxysuccinimide esters, hydroxysulfosuccinimide esters, imido esters, hydrazines, azidonitrophenyl, azides, 3-(2-pyridyl dithio)-propionamide and glyoxal. Additional suitable reactive moieties for use in fluorescent labels are described in U.S. Patent No. 5,268,486 which is incorporated herein by reference.
  • Fluorescent dyes commonly have an absorbance range of between about 300 and 900 nm and preferably have a Stokes shift of at least about 20 nm. Fluorescent dyes that absorb in the 500 to 900 nm range are preferred because they are spectrally removed from other components that may be present in a biological sample and because they may be used with inexpensive light sources. Fluorescent dyes that have a high extinction coefficient and a high quantum yield are also preferred. Fluorescent dyes used for labeling biomolecules, such as carbohydrates, proteins and DNA, are preferably water soluble since the biomolecules to be labeled generally have limited solubility in nonaqueous solvents.
  • the fluorescent dye is photostable. However, dyes with a fluoresence absorbance greater than 500 nm tend to be less photostabile. Fluorescent dyes also should not be prone to aggregation. Dye aggregation, also known as “stacking" increases the frequency of fluorescence quenching which reduces the strength of the fluoresence signal observed. Most fluorescent dyes are large planar molecules, are intrinsically hydrophobic and therefore have a tendency to aggregate or "stack," especially in aqueous solutions. Dyes with a fluoresence absorbance greater than 500 nm generally have a greater tendency to stack due to their increased size and associated lower solubility. Non-aggregating, photostable fluorescent dyes with a fluoresence absorbance greater than 500 nm are therefore needed.
  • Fluorescent probes are particularly prone to stack in high salt solutions and when in high local concentrations on protein surfaces.
  • tetramethylrhodamine a commonly used laser dye, produces protein-dye conjugates which predominantly consist of the aggregated dye. Aggregated dyes appear blue-shifted by visible absorbance spectra.
  • Amino-substituted cyanine dyes such as IR144, are prone to aggregation in aqueous solutions, even in low-salt solutions (i.e. 0.1 M NaCl). Non-aggregated amino-substituted cyanine dyes have only been found to exist in organic solvents.
  • the absorbance spectra of protein-dye conjugates can be simulated by obtaining spectra of the dye in high salt solutions (e.g. 4 M NaCl).
  • Dye aggregation may be minimized by constructing highly ionic dyes such as arylsulfonates taught in U.S. Patent No. 5,268,486 or by using naturally occurring fluorescent probes such as phycobiliproteins.
  • the present invention relates to cyanine dyes substituted with either an N-heteroaromatic ion or an iminium ion which have a fluoresence absorbance of between about 500 and 900 nm, a reduced tendency to aggregate and enhanced photostability.
  • the cyanine dyes of the present invention are represented by the formula
  • R, and R 2 are taken together to form an aromatic ring or a fused polycyclic aromatic ring;
  • R 3 and R are taken together to form an aromatic ring or a fused polycyclic aromatic ring
  • R 5 and R- are independently selected from the group consisting of (CH 2 ) p X where p is 1-18 and X is a functional group that reacts with amino, hydroxy or sulfhydryl nucleophiles;
  • R 7 and R 8 are independently selected from the group consisting of hydrogen, C 1 -CIO alkyl groups and where R 7 and R g are taken together to form a five- or six- membered heterocyclic ring;
  • Ro are each independently selected from the group consisting of hydrogen, alkyl and where more than one R ⁇ are taken together to form a five- or six- membered ring;
  • Y is selected from the group consisting of C(CH 3 ) 2 , S, O and Se;
  • the present invention also relates to a method for using the cyanine dyes of the present invention for fluorescence labeling molecules, particularly biomolecules such as antibodies, DNA, carbohydrates and cells.
  • Figure 1 depicts the absorbance spectrum of a BHDMAP-protein conjugate.
  • Figure 2A depicts the spectra of BHCI in low (0.1 M NaCl, 50 mM phosphate, pH 7) and high (3.8 M NaCl, 50 mM phosphate, pH 7) salt solutions.
  • Figure 2B depicts the spectra of BHDMAP in low (0.1 M NaCl, 50 mM phosphate, pH 7) and high (3.8 M NaCl, 50 mM phosphate, pH 7) salt solutions.
  • Figure 3 depicts the spectra of IR144 in a low salt solution (0.1 M NaCl,
  • Figure 4 depicts the photodecomposition rates of several cyanine dyes.
  • Cy5 and Cy7 are arylsulfonate dyes of U.S. Patent No. 5,268,486.
  • the present invention relates to a class of cyanine dyes substituted with either an N-heteroaromatic ion or an iminium ion having a fluorescence absorbance between about 500 and 900 nm.
  • This class of cyanine dyes have the advantage of being photostable and are not prone to aggregation.
  • the present invention also relates to a method for fluorescence labeling molecules using the substituted cyanine dyes of the present invention as fluorescent probes.
  • N-heteroaromatic ion and iminium ion substituted cyanine dyes of the present invention are represented by the formula:
  • R, and R 2 are taken together to form an aromatic ring or a fused polycyclic aromatic ring;
  • R 3 and R are taken together to form an aromatic ring or a fused polycyclic aromatic ring
  • R 5 and R are independently selected from the group consisting of (CH 2 ) p X where p is 1-18 and X is a functional group that reacts with amino, hydroxy and sulfhydryl nucleophiles;
  • R 7 and R 8 are independently selected from the group consisting of hydrogen, Cl -CIO alkyl groups and where R 7 and R 8 are taken together to form a five- or six- membered heterocyclic ring;
  • Ro are each independently selected from the group consisting of hydrogen, alkyl and where more than one Ro are taken together to form a five- or six- membered ring;
  • Y is selected from the group consisting of C(CH 3 ) 2 , S, O and Se; and Z is selected from the group consisting of C(CH 3 ) 2 , S, O and Se.
  • R, - R 2 and R 3 - R are both preferably taken together to form a benzene or naphthalene ring.
  • the aromatic ring or fused polycyclic aromatic rings formed by R, and R 2 taken together and R 3 and R 4 taken together may be either unsubstituted or substituted. Substitution of the aromatic ring or rings with electron donating groups, such as primary, secondary and tertiary alkyl groups, may be used to lower the absorbance wavelength of the dye relative to an unsubstituted dye.
  • substitution of the aromatic ring or rings with electron withdrawing groups such as nitro, cyanate, acid, halide, alkoxy, aryloxy, ester, ether, sulfide, thioether, alcohol, alkene, alkyne and aryl groups, may be used to increase the absorbance wavelength of the dye relative to an unsubstituted dye.
  • the reactive moieties (X) employed with R 5 and R- may be any functional group that reacts with the amino, hydroxy and/or sulfhydryl nucleophiles commonly found on the carbohydrates, proteins, DNA or cells to be labeled by the fluorescent dye.
  • Suitable reactive moieties include, but are not limited to, carboxylic acids, acid halides, sulfonic acids, esters, aldehydes, disulfides, isothiocyanates, isocyanates, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridines, mono- or di- halogen substituted diazines, maleimide, aziridines, sulfonyl halides, hydroxysuccinimide esters, hydroxysulfosuccinimide esters, imido esters, hydrazines, azidonitrophenyl, azides, 3-(2-pyridyl dithio)-propionamide and glyoxal. Additional suitable reactive moieties for use in fluorescent labels are described in U.S. Patent No. 5,268,486.
  • the reactive moieties used in R 5 and Rg are preferably succidimidyl esters.
  • R 7 and R 8 are preferably taken together to form a heterocyclic five- or six- membered ring including, for example, pyridinium, imidazolium, pyrrolium, pyrazolium, pyrazinium, pyrimidinium, pyridazinium, quinolinium, purinium and isoquinolinium.
  • R 7 and R 8 are more preferably taken together to form a pyridinium or an imidazolium ring.
  • R 7 and R 8 are most preferably taken together to form a 4-dimethylaminopyridium, 4-(4-morpholinyl) pyridinium, or a 1-methylimidazolium substituent.
  • the heterocyclic ring formed by R 7 and R 8 taken together may be substituted or unsubstituted.
  • R 7 and R 8 may be further substituted by either electron donating or electron withdrawing groups in electron communication with the aromatic system of the dye in order to influence the fluorescence absorbance wavelength of the dye.
  • Substitution of R 7 and R 8 with electron donating groups, such as primary, secondary and tertiary alkyl groups, may be used to decrease the fluorescence absorbance wavelength of the dye relative to where R 7 and R 8 are substituted with hydrogen.
  • substitution of R 7 and R 8 with electron withdrawing groups such as nitro, cyanate, acid, halide, alkoxy, aryloxy, ester, ether, sulfide, thioether, alcohol, alkene, alkyne and aryl groups, may be used to increase the fluorescence absorbance wavelength of the dye relative to where R 7 and R 8 are substituted with hydrogen.
  • electron withdrawing groups such as nitro, cyanate, acid, halide, alkoxy, aryloxy, ester, ether, sulfide, thioether, alcohol, alkene, alkyne and aryl groups
  • the Ro substituents are preferably selected such that the carbon atoms situated a and a' to the iminium ion form part of either a five- or six- membered ring.
  • the five- or six- membered ring may be substituted or unsubstituted.
  • Y and Z may be either C(CH 3 ) 2 , S, O or Se.
  • Y and Z are C(CH 3 ) 2 .
  • Y and Z serve to keep the cyanine dye relatively planar and provide the dye with fluorescence.
  • a preferred subclass of cyanine dyes of the present invention includes those cyanine dyes of the formula
  • R b R 2 , R 3 , R,, R 5 , Rg, R 7 , R 8 , Y and Z are as specified above and wherein r is either 1, 2 or 3.
  • Table 1 provides the names, structures, absorbance and fluorescence emission wavelengths of several cyanine dyes of the present invention and of their chloro-substituted precursors. NHC1 and ZFHC1 do not have an absorbance maximum in phosphate buffered saline (PBS).
  • PBS phosphate buffered saline
  • the cyanine dyes of the present invention have been found to possess enhanced photostability and are not prone to aggregation. Without being bound by theory, it is believed that the N-heteroaromatic ion and the iminium ion inhibits aggregation of these dyes. In addition to inhibiting aggregation, the N- heteroaromatic ion and the iminium ion are also believed to contribute to the photostability of these dyes.
  • the reaction mixture contained starting dye (17%), monoester (60%) and diester (23%).
  • Acetic acid (20 ⁇ L) and methanol (0.5 mL) were added and the solution filtered to remove dicyclohexylurea.
  • the solution was concentrated to dryness and redissolved in DMF (0.5 mL).
  • the concentration of the succinimidyl ester solution was determined by dilution of an aliquot into phosphate buffered saline and measurement of the optical density at 786 nm. The extinction coefficient was assumed to be 200,000 cm "'M '1 .
  • the concentration of BHDMAP succinimidyl ester was found to be 12 mg/mL. 4. Antibody labeling with BHDMAP.
  • the tendency of dyes to aggregate on proteins can be simulated by measuring the absorbance of the dye in low and high salt solutions.
  • High salt solutions simulate the environment of the dye in high local concentration on the surface of a protein.
  • Figure 2A shows the spectra of BHCI in low (0.1 M NaCl, 50 mM phosphate, pH 7) and high (3.8 M NaCl, 50 mM phosphate, pH 7) salt solutions.
  • Figure 2B shows the spectra of BHDMAP in low (0.1 M NaCl, 50 mM phosphate, pH 7) and high (3.8 M NaCl, 50 mM phosphate, pH 7) salt solutions.
  • BHCI appears to aggregate even in low salt, and in high salt the absorbance maximum has shifted to shorter wavelength.
  • FIG. 1 shows the spectra of a related dye, IR144, in low salt and in dimethylformamide solutions. Based on the spectra shown in Figure 3, IR144 appears to aggregate in low salt solutions. 6. Photodecomposition of dyes.
  • Cy5 and Cy7 are the penta- and hepta-methine derivatives, respectively, of a class of arylsulfinate dyes described in U.S. Patent No. 5,268,486.
  • the concentration of the dye was assumed to be proportional to the optical density of the dye solution in accordance with Beer's Law.
  • a plot of the logarithm of the normalized absorbance (In[Dye]/[Dye] 0 ) vs. time is shown in Figure 4. Values of k were determined from least squares analysis and the half-life of each dye determined.
  • the most stable dye was found to be the dye with the shortest wavelength, Cy5, whose structure contains five methine groups. The remaining dyes contain seven methine groups. BHDMAP, NHMI and Cy7 all have similar stabilities. The least stable dye was found to be BHCI. Substitution of the chloride with dimethylaminopyridme to provide BHDMAP was found to improve the photostability of the cyanine dye seven-fold.

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Abstract

Cette invention se rapporte à des colorants à base de cyanine substitués par des ions iminium, ayant une absorbance de fluorescence comprise entre 500 et 900 nm, une tendance réduite à former des agrégats et une photostabilité améliorée. Ces colorants à base de cyanine sont représentés par la formule (I), où: n vaut 0, 1, 2 ou 3; m vaut 0, 1, 2 ou 3; R1 et R2 forment ensemble un cycle aromatique ou un cycle aromatique polycyclique fusionné; R3 et R4 forment ensemble un cycle aromatique ou un cycle aromatique polycyclique fusionné; R5 et R6 sont choisis séparément dans le groupe constitué par (CH2)pX, où p vaut 1 à 18, et X représente un groupe fonctionnel qui réagit avec amino, hydroxy et sulfhydryl nucléophiles, R7 et R8 sont choisis séparément dans le groupe constitué par hydrogène, des groupes alkyle C1-C10 et où R7 et R8 forment ensemble un cycle hétérocyclique à 5 ou 6 éléments; tous les R9 sont choisis séparément dans le groupe constitué par hydrogène, par alkyle et où plus d'un R9 forment ensemble un cycle à 5 ou 6 éléments; Y est choisi dans le groupe constitué par C(CH3)2, S, O et Se; et Z est choisi dans le groupe constitué par C(CH3)2, S, O et Se. Cette invention se rapporte également à un procédé pour utiliser ces colorants à base de cyanine afin d'obtenir un marquage fluorescent sur des molécules, en particulier des biomolécules telles que des anticorps, des ADN, des hydrates de carbone et des cellules.
PCT/US1995/008778 1994-06-30 1995-06-29 Colorants a base de cyanine substitues par des ions n-heteroaromatiques et par des ions iminium, utilises comme marques fluorescentes WO1996000902A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP95926272A EP0769145A1 (fr) 1994-06-30 1995-06-29 Colorants a base de cyanine substitues par des ions n-heteroaromatiques et par des ions iminium, utilises comme marques fluorescentes
AU30085/95A AU3008595A (en) 1994-06-30 1995-06-29 N-heteroaromatic ion and iminium ion substituted cyanine dyes for use as fluorescence labels

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US08/268,852 1994-06-30
US08/268,852 US5453505A (en) 1994-06-30 1994-06-30 N-heteroaromatic ion and iminium ion substituted cyanine dyes for use as fluorescence labels
US38860795A 1995-02-14 1995-02-14
US08/388,607 1995-02-14

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WO1996000902A1 true WO1996000902A1 (fr) 1996-01-11

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CA (1) CA2194150A1 (fr)
WO (1) WO1996000902A1 (fr)

Cited By (25)

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WO1998015829A1 (fr) * 1996-10-07 1998-04-16 Amersham International Plc Analyse de glucides
US5755943A (en) * 1984-03-29 1998-05-26 Li-Cor, Inc. DNA sequencing
US5800995A (en) * 1984-03-29 1998-09-01 Li-Cor, Inc. Sequencing near infrared and infrared fluorescence labeled DNA for detecting using laser diodes and suitable labels therefor
US5863403A (en) * 1984-03-29 1999-01-26 The Board Of Regents Of The University Of Nebraska Digital DNA typing
WO1999005221A1 (fr) * 1997-07-28 1999-02-04 Nycomed Amersham Plc Colorants de cyanine
US6004446A (en) * 1984-03-29 1999-12-21 Li-Cor, Inc. DNA Sequencing
US6086737A (en) * 1984-03-29 2000-07-11 Li-Cor, Inc. Sequencing near infrared and infrared fluorescence labeled DNA for detecting using laser diodes and suitable labels therefor
WO2000063418A1 (fr) * 1999-04-15 2000-10-26 The Regents Of The University Of California Dosages de transport proteidique
US6143151A (en) * 1984-03-29 2000-11-07 Li-Cor, Inc. DNA sequencing
US6207421B1 (en) 1984-03-29 2001-03-27 Li-Cor, Inc. DNA sequencing and DNA terminators
EP1163372A1 (fr) * 1999-03-24 2001-12-19 Princeton Separations Colorants a base de cyanine rigidifies dans un plan et chimiquement reactifs, et derives de ces derniers
EP1250091A1 (fr) * 2000-01-18 2002-10-23 Mallinckrodt Inc. Colorants hydrophiles a base de cyanine
EP1326647A1 (fr) * 2000-10-16 2003-07-16 Mallinckrodt Inc. Composes photosensibles destines a determiner instantanement la fonction d'un organe
JP2004513893A (ja) * 2000-10-16 2004-05-13 マリンクロッド・インコーポレイテッド 器官機能モニタリング用の新規染料
EP1443860A2 (fr) * 2001-10-17 2004-08-11 Mallinckrodt Inc. Colorants carbocyanine pour applications photodiagnostiques et therapeutiques combinees
JP2004526669A (ja) * 2000-10-16 2004-09-02 マリンクロッド・インコーポレイテッド 最小の生理機能モニタリング物質としてのインドール化合物
EP1606351A2 (fr) * 2003-03-21 2005-12-21 Amersham Biosciences Corp. Reactifs de marquage constitues de colorants de cyanine a meso-substitution
US6995262B1 (en) 1999-08-05 2006-02-07 Bayer Cropscience Ag Use of acylsulfonamido-substituted polymethine dyes as fluorescene dyes and/or markers
US7556797B2 (en) * 2000-10-16 2009-07-07 Mallinckrodt Inc. Minimally invasive physiological function monitoring agents
US7767194B2 (en) 2000-01-18 2010-08-03 Mallinckrodt Inc. Optical diagnostic and therapeutic agents and compositions
US7767829B2 (en) * 2005-04-22 2010-08-03 Ge Healthcare Uk Limited Water-soluble fluoro-substituted cyanine dyes as reactive fluorescence labelling reagents
US8628753B2 (en) 2008-03-28 2014-01-14 Emory University Reduced dye probes for the detection of radical oxygen species
US9201014B2 (en) 2010-03-24 2015-12-01 National University Of Singapore Development of photostable near-IR cyanine dyes for in vivo imaging
JP2019172818A (ja) * 2018-03-28 2019-10-10 日本化薬株式会社 シアニン化合物
WO2023235452A1 (fr) * 2022-06-01 2023-12-07 Georgia State University Research Foundation, Inc. Colorants de cyanine d'ammonium quaternaire

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US5268486A (en) * 1986-04-18 1993-12-07 Carnegie-Mellon Unversity Method for labeling and detecting materials employing arylsulfonate cyanine dyes
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Cited By (36)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6207421B1 (en) 1984-03-29 2001-03-27 Li-Cor, Inc. DNA sequencing and DNA terminators
US5755943A (en) * 1984-03-29 1998-05-26 Li-Cor, Inc. DNA sequencing
US5800995A (en) * 1984-03-29 1998-09-01 Li-Cor, Inc. Sequencing near infrared and infrared fluorescence labeled DNA for detecting using laser diodes and suitable labels therefor
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