公开(公告)号：US5534416A

公开(公告)日：1996-07-09

申请号：US148847

申请日：1993-11-08

申请人： Paul J. Millard ,  Bruce L. Roth ,  Stephen T. Yue ,  Richard P. Haugland

发明人： Paul J. Millard ,  Bruce L. Roth ,  Stephen T. Yue ,  Richard P. Haugland

IPC分类号： C07D215/22 ,  C07D211/76 ,  C07D213/60 ,  C07D213/61 ,  C07D213/64 ,  C07D215/16 ,  C07D215/18 ,  C07D215/227 ,  C07D277/64 ,  C07D413/06 ,  C07D417/06 ,  C07D417/14 ,  C09B23/00 ,  C09B23/02 ,  C09B23/04 ,  C09B69/06 ,  C12Q1/68 ,  G01N21/64 ,  G01N21/78 ,  G01N27/26 ,  G01N31/22 ,  G01N33/48 ,  G01N33/50 ,  G01N33/52 ,  G01N33/58 ,  G01N33/00 ,  C07H1/00 ,  C12Q1/04

CPC分类号： C07D211/76 ,  C07D215/18 ,  C07D215/227 ,  C07D417/06 ,  C07D417/14 ,  C09B23/02 ,  Y10S436/80 ,  Y10T436/143333

摘要： The invention relates to a method of analyzing the viability of a sample of cells using an aqueous solution comprising two fluorescent dyes. Dye I has the formula: ##STR1## where R.sup.2 is C.sub.1-6 alkyl; Z.sup.- is a biologically compatible counterion;X is O; S; Se; or NR.sup.15, where R.sup.15 is H or C.sub.1-6 alkyl; or CR.sup.16 R.sup.17, where R.sup.16 and R.sup.17, which may be the same or different, are independently H or C.sub.1-6 alkyl, or the carbons of R.sup.16 and R.sup.17 taken in combination complete a five or six membered saturated ring; and the benzazolium is optionally further substituted;n=0, 1, or 2;Y is --CR.sup.3 .dbd.CR.sup.4 --; p and m=0 or 1, such that p+m=1;R.sup.5 is a C.sub.1-6 alkyl, C.sub.1-6 alkenyl, C.sub.1-6 polyalkenyl, C.sub.1-6 alkynyl or C.sub.1-6 polyalkynyl group; or R.sup.5 is an OMEGA;R.sup.3, R.sup.4, R.sup.6 and R.sup.7, which may be the same or different, are independently H; or a C.sub.1-6 alkyl, C.sub.1-6 alkenyl, C.sub.1-6 polyalkenyl, C.sub.1-6 alkynyl or C.sub.1-6 polyalkynyl group; or halogen; or --OR.sup.8, --SR.sup.8, --(NR.sup.8 R.sup.9), where R.sup.8 and R.sup.9, which may be the same or different, are independently H; or alkyl groups having 1-6 carbons; or 1-2 substituted or unsubstituted alicyclic, heteroalicyclic, aromatic, or heteroaromatic rings, containing 1-4 heteroatoms, wherein the heteroatoms are O, N, or S; or R.sup.8 and R.sup.9 taken in combination are --(CH.sub.2).sub.2 --L--(CH.sub.2).sub.2 -- where L=--O--, --NR.sup.10 --, --CH.sub.2 -- or a single bond where R.sup.10 is H or an alkyl group having 1-6 carbons; or --OSO.sub.2 R.sup.19 where R.sup.19 is C.sub.1-6 alkyl, or C.sub.1-6 perfluoroalkyl, or aryl; or an OMEGA; or R.sup.6 and R.sup.7, taken in combination are --(CH.sub.2).sub.v -- where v=3 or 4, or R.sup.6 and R.sup.7 form a fused aromatic ring that is optionally further substituted;such that at least one of R.sup.3, R.sup.4, R.sup.5, R.sup.6 and R.sup.7, or a substituent on the aromatic ring formed by R.sup.6 and R.sup.7, is an OMEGA; where OMEGA is a cyclic substituent that is attached by a single bond.Fluorescent Dye II selectively stains either viable or non-viable cells with a detectable fluorescent response that is different from the fluorescent response of Dye I. The stained cells are illuminated at a suitable absorption wavelength, and the fluorescent response is detected to distinguish viable and non-viable cells based on the fluorescent response.

公开(公告)号：US5501980A

公开(公告)日：1996-03-26

申请号：US247013

申请日：1994-05-20

申请人： Haralambos E. Katerinopoulos ,  Helene Iatridou ,  Evangelia Foukaraki ,  Mohammad N. Malekzadeh ,  Michael A. Kuhn ,  Richard P. Haugland

发明人： Haralambos E. Katerinopoulos ,  Helene Iatridou ,  Evangelia Foukaraki ,  Mohammad N. Malekzadeh ,  Michael A. Kuhn ,  Richard P. Haugland

IPC分类号： C07D417/04 ,  C07D417/14 ,  C09K11/06 ,  G01N31/22 ,  G01N33/18 ,  G01N33/20

CPC分类号： C07D417/14 ,  C07D417/04 ,  C09K11/06 ,  G01N31/22 ,  G01N33/1813

摘要： The indicator compounds of the present invention are substituted or unsubstituted BAPTA-type 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 Ca.sup.2+.The compounds of the invention have the core structure ##STR1## or the structure ##STR2## where m=2 or 3 and X can be S, O, or C(CH.sub.3).sub.2. The above core structures are 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.

摘要翻译： 本发明的指示剂化合物是含有苯并唑基 - 香豆素亚结构的取代或未取代的BAPTA型螯合剂及其药学上可接受的无毒盐和酯。 这些化合物可用于阳离子金属离子，特别是Ca 2+的检测和定量。 本发明的化合物具有核心结构“IMAGE”或结构“IMAGE”，其中m = 2或3，X可以是S，O或C（CH 3）2。 上述核心结构任选地被改变指示剂的结合亲和力，转移指示剂的光谱性质或用作制备各种缀合物的反应位点的取代基取代。

公开(公告)号：US5453517A

公开(公告)日：1995-09-26

申请号：US843360

申请日：1992-02-25

申请人： Michael A. Kuhn ,  Richard P. Haugland

发明人： Michael A. Kuhn ,  Richard P. Haugland

IPC分类号： C07D209/14 ,  C07D311/82 ,  C07D311/90 ,  C07D413/04 ,  C09K11/06 ,  C07D311/88 ,  C07D265/30 ,  C07H23/00 ,  C08B37/02

CPC分类号： C07D311/82 ,  C07D209/14 ,  C07D311/90 ,  C07D413/04 ,  C09K11/06

摘要： 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. gel and resin polymers, polysaccharides, polypeptides, nucleic acids, and liposomes); or combinations thereof.

摘要翻译： 本发明涉及下式的1,2-二 - （2-氨基苯氧基乙烷）-N，N，N'，N'-四乙酸（BAPTA）的荧光和/或活性衍生物，其中至少一个 的W和X是具有或不具有间隔基的官能团，其终止于醇或苯酚，硫醇，卤代乙酰胺，烷基卤，胺或苯胺，羧酸，酸酐，异氰酸酯，异硫氰酸酯 ，马来酰亚胺或活化酯。 BAPTA样分子可以通过具有或不具有间隔基的另外的官能团或通过CH 3，NO 2，CF 3，F，Cl，Br，I或其羧酸衍生物或其药学上可接受的盐进一步取代一次或多次，或 通过吲哚基或苯并呋喃荧光团。 官能团允许一个或多个氧杂环荧光团（例如荧光素，香豆素，罗丹明）的随后共价连接。 或多分子组件（例如凝胶和树脂聚合物，多糖，多肽，核酸和脂质体）; 或其组合。

公开(公告)号：US5451663A

公开(公告)日：1995-09-19

申请号：US45758

申请日：1993-04-08

申请人： Hee C. Kang ,  Richard P. Haugland

发明人： Hee C. Kang ,  Richard P. Haugland

IPC分类号： C07F5/02 ,  C07H13/04 ,  C07H19/10 ,  C07H21/00 ,  C09B57/00 ,  C09K11/06 ,  G01N33/533 ,  C07K17/00 ,  C07H19/00 ,  C07K14/00 ,  C07K16/00

CPC分类号： C07H13/04 ,  C07F5/022 ,  C07H19/10 ,  C07H21/00 ,  C09B57/00 ,  C09K11/06 ,  G01N33/533

摘要： 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.

摘要翻译： 本发明涉及二吡咯亚甲基二氟化物荧光染料的衍生物，其在波长比525nm波长处具有最大吸收，并且与核酸，蛋白质，碳水化合物和其它生物衍生或合成的化学物质化学反应以形成染料缀合物。 染料缀合物通常具有以下结构：其中至少一个取代基R 1 -R 7与特定结合对成员共价结合，此外，其中至少一个取代基R 1 -R 7含有红色部分，其为 杂芳基或烯基。 其余取代基可以相同或不同，是氢，卤素，烷基（含1-5个碳原子），芳基，芳基烷基或磺基。

公开(公告)号：US5445946A

公开(公告)日：1995-08-29

申请号：US206081

申请日：1994-03-03

申请人： Bruce L. Roth ,  Paul J. Millard ,  Stephen T. Yue ,  Richard P. Haugland

发明人： Bruce L. Roth ,  Paul J. Millard ,  Stephen T. Yue ,  Richard P. Haugland

IPC分类号： C07D211/76 ,  C07D215/18 ,  C07D215/22 ,  C07D215/227 ,  C07D417/06 ,  C07D417/14 ,  C09B23/02 ,  C12Q1/04 ,  C07H17/00 ,  C12Q1/02 ,  C12Q1/68

CPC分类号： C07D211/76 ,  C07D215/18 ,  C07D215/227 ,  C07D417/06 ,  C07D417/14 ,  C09B23/02

摘要： 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.sup.10 R.sup.11 where R.sup.10 and R.sup.11, which may be the same or different, are independently alkyl groups having 1-6 carbons, or R.sup.10 and R.sup.11 taken in combination complete a five or six membered saturated ring;R.sup.3, R.sup.4 and R.sup.5, which may be the same or different, are independently H; or an alkyl, alkenyl, polyalkenyl, alkynyl or polyalkynyl group having 1-6 carbons; or a halogen; or --OH, --OR.sup.6, --SR.sup.6 or --(NR.sup.6 R.sup.7); or R.sup.4 and R.sup.5, taken in combination form a quinolinium ring system that is optionally similarly substituted or unsubstituted;Z.sup.- is a biologically compatible counterion;L is a halogen or good leaving group; andOMEGA is a saturated or unsaturated, substituted or unsubstituted, cyclic substituent.

摘要翻译： 本发明涉及形成与真菌细胞独特的独特的血管内结构的方法，并且可以与细胞活力相关。 本发明优选的染料用下式描述：其中每个R 1独立地是H; 或具有1-6个碳原子的烷基; 或三氟甲基; 或卤素; 或-OR 6，-SR 6或 - （NR 6 R 7）其中R 6和R 7可以相同或不同，独立地为H; 或具有1-6个碳的烷基; 或1-2个含有1-4个杂原子的脂环族，杂脂环，芳族或杂芳环，其中杂原子是O，N或S; 或R 6和R 7组合是 - （CH 2）2-M-（CH 2）2 - ，其中M =单键，-O-，-CH 2 - 或-NR 8 - ，其中R 8是H或具有1- 6个碳; 和t = 1-4; R2是具有1-6个碳原子的烷基; X是O，S，Se或NR 9，其中R 9是H或具有1-6个碳的烷基; 或X为CR10R11，其中R10和R11可以相同或不同，独立地为具有1-6个碳原子的烷基，或R10和R11组合完成一个五或六元饱和环; R 3，R 4和R 5可以相同或不同，独立地为H; 或具有1-6个碳原子的烷基，链烯基，聚链烯基，炔基或聚炔基; 或卤素; 或-OH，-OR 6，-SR 6或 - （NR 6 R 7）; 或R 4和R 5组合形成任选相似取代或未取代的喹啉鎓环系; Z-是一种生物相容的抗衡离子; L是卤素或良好的离去基团; 并且OMEGA是饱和或不饱和的，取代或未取代的环状取代基。

公开(公告)号：US5362628A

公开(公告)日：1994-11-08

申请号：US26633

申请日：1993-03-05

申请人： Richard P. Haugland ,  Yu-Zhong Zhang ,  Ram Sabnis ,  Nels A. Olson ,  John J. Naleway ,  Rosaria P. Haugland

发明人： Richard P. Haugland ,  Yu-Zhong Zhang ,  Ram Sabnis ,  Nels A. Olson ,  John J. Naleway ,  Rosaria P. Haugland

IPC分类号： C12Q1/04 ,  C12Q1/34 ,  G01N33/535 ,  C12Q1/00 ,  A61K37/02 ,  C07H15/24 ,  G01N33/40

CPC分类号： C12Q1/04 ,  C12Q1/34 ,  G01N33/535 ,  C12Q2334/00 ,  C12Q2334/20 ,  C12Q2334/40 ,  C12Q2337/20 ,  G01N2333/924 ,  Y10S530/802

摘要： 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--).After the substrate enters the cells, the analyte removes BLOCK to make REPORTER detectable by the change in spectral properties, and the haloalkyl XR reacts with the intracellular thiol to form the thioether conjugate inside the cells, which is well-retained in the cells.

摘要翻译： 本发明提供了通过仅在存在特定酶的完整细胞中改善可检测报道分子的保留来分析细胞中的代谢活性的方法。 特别地，涉及将报道分子的卤代烷基取代的衍生物与含细胞内半胱氨酸的肽结合的两部分方法的改进的保留结果，同时解除报道分子的阻断。 用于分析细胞代谢活性的方法涉及使用具有XR-REPORTER-BLOCK形式的底物，其中-BLOCK是通过特定分析物的作用被选择为可去除的基团，以产生与底物的不同的REPORTER光谱性质 -REPORTER-是一个分子，当不再通过BLOCK-REPORTER键与BLOCK结合时，具有与底物不同的光谱性质，XR-是可以与胞内硫醇（ZSH）共价反应的卤代烷基部分， 以形成硫醚缀合物（ZSR-）。 在底物进入细胞后，分析物除去BLOCK以通过光谱性质的变化使REPORTER可检测到，并且卤代烷基XR与细胞内硫醇反应以在细胞内形成硫醚缀合物，其在细胞中良好保留。

公开(公告)号：US5314805A

公开(公告)日：1994-05-24

申请号：US783182

申请日：1991-10-28

申请人： Richard P. Haugland ,  Ian C. MacCoubrey ,  Patrick L. Moore

发明人： Richard P. Haugland ,  Ian C. MacCoubrey ,  Patrick L. Moore

IPC分类号： C12Q1/02 ,  C12Q1/04 ,  C12Q1/68

CPC分类号： C12Q1/02 ,  C12Q1/025 ,  Y10S435/968 ,  Y10S436/80

摘要： 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.

摘要翻译： 本发明涉及一种使用两种荧光试剂同时检测活细胞和死细胞的方法：钙黄绿素AM和同源二聚体。 活细胞被钙黄绿素AM酶促水解产生的强烈均匀的绿色荧光区分开来：死亡或损伤的细胞的特征在于用同源二聚体染色的核酸产生的明亮的红色荧光： 确定细胞活力并监测细胞毒性剂或事件。

公开(公告)号：US5274113A

公开(公告)日：1993-12-28

申请号：US786767

申请日：1991-11-01

申请人： Hee C. Kang ,  Richard P. Haugland

发明人： Hee C. Kang ,  Richard P. Haugland

IPC分类号： C07F5/02 ,  C07H13/04 ,  C07H19/10 ,  C07H21/00 ,  C09B57/00 ,  C09K11/06 ,  G01N33/533 ,  C07D207/44 ,  H01S3/20

CPC分类号： C07H13/04 ,  C07F5/022 ,  C07H19/10 ,  C07H21/00 ,  C09B57/00 ,  C09K11/06 ,  G01N33/533

摘要： 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.

摘要翻译： 本发明涉及二吡咯亚甲基二氟化物荧光染料的衍生物，其在波长比525nm波长处具有最大吸收，并且与核酸，蛋白质，碳水化合物和其它生物衍生或合成的化学材料具有化学反应性。 染料通常具有以下结构：其中至少一个取代基R 1 -R 7是反应性官能团，并且至少一个取代基R 1 -R 7含有红色部分。 红色部分是不饱和有机基团，优选杂芳基或烯基。 其余取代基可以相同或不同，是氢，卤素，烷基（含1-5个碳原子），芳基，芳基烷基或磺基。 这些染料用于制备与配体或受体的特异性结合对成员的新型缀合物。

公开(公告)号：US5132432A

公开(公告)日：1992-07-21

申请号：US410837

申请日：1989-09-22

申请人： Richard P. Haugland ,  James E. Whitaker

发明人： Richard P. Haugland ,  James E. Whitaker

IPC分类号： C07C309/43 ,  C07D207/16 ,  C07D207/40 ,  C07D207/404 ,  C09B57/00

CPC分类号： C07D207/404 ,  C07C309/43 ,  C07D207/16 ,  C09B57/001 ,  C07C2103/50

摘要： The invention relates to novel fluorescent dyes based on the following pyrenyloxy sulfonic acid structure: ##STR1## wherein at least one of R.sub.1, R.sub.2, R.sub.3, and R.sub.4 is --OCH.sub.2 C(O)OCH.sub.3, --OCH.sub.2 CON(O)H, --OCH.sub.2 CON.sub.3, --OCH.sub.2 CONHNH.sub.2, ##STR2## or --CH.sub.2 CONH(CH.sub.2).sub.n NH.sub.2 where n is an integer from 1 to 10; at least one of R.sub.1, R.sub.2, R.sub.3, and R.sub.4 is sulfonic acid or alkali salts of sulfonic acid; and all other of R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are hydrogen or a hydroxyl group. The new reagents contain functional groups typically found in biomolecules or polymers. The spectral properties of the fluorescent dyes are sufficiently different in wavelengths and intensity from fluorescein as to permit simultaneous use of both dyes with minimum interference. The dyes have narrower spectral band-widths and smaller Stokes' shifts than other reactive dyes with similar spectral properties, do not show appreciable sensitivity to pH, have higher solubilities in aqueous solvents and have good quantum yields and photostability.

摘要翻译： 本发明涉及基于以下吡喃氧基磺酸结构的新型荧光染料：其中R 1，R 2，R 3和R 4中的至少一个为-OCH 2 C（O）OCH 3，-OCH 2 CON（O）H，-OCH 2 CON 3 ，-OCH 2 CONHNH 2，或-CH 2 CONH（CH 2）n NH 2，其中n为1至10的整数; R 1，R 2，R 3和R 4中的至少一个是磺酸或磺酸的碱金属盐; 并且R 1，R 2，R 3和R 4中的所有其它基团是氢或羟基。 新试剂含有通常存在于生物分子或聚合物中的官能团。 荧光染料的光谱性质在荧光素的波长和强度方面是充分不同的，以允许以最小干扰同时使用两种染料。 染料具有较窄的光谱带宽和比具有相似光谱性质的其它活性染料更小的斯托克斯位移，对pH具有明显的敏感性，在水性溶剂中具有较高的溶解度并且具有良好的量子产率和光稳定性。

公开(公告)号：US4945171A

公开(公告)日：1990-07-31

申请号：US83459

申请日：1987-08-10

申请人： Richard P. Haugland ,  James Whitaker

发明人： Richard P. Haugland ,  James Whitaker

IPC分类号： C07D493/10 ,  C09B11/28 ,  G01N33/84

CPC分类号： C07D493/10 ,  C09B11/28 ,  G01N33/84

摘要： Synthesis and applications of fluorescent dyes which are derivatives of benzo[c]xanthenes is described. The dyes exhibit pH dependent absorption and fluorescence spectra with pKas near the normal physiological range. Unlike fluorescein, the dyes exhibit emission of different characteristic wavelengths dependent on the pH of the medium. This permits several methods of measuring the pH of the medium in contact with the indicator including measuring two emissions with one excitation, selectively exiting the acid and base forms independently and measuring their emission at either single or dual wavelengths, or measuring the characteristic pH dependent absorption or fluorescence excitation spectral. Methods are presented for making the indicators permeant to cell membranes for the measurement of intracellular pH.

摘要翻译： 描述了作为苯并[c]呫吨的衍生物的荧光染料的合成和应用。 染料表现出pH依赖性吸收和荧光光谱，pKas在正常生理范围附近。 与荧光素不同，染料表现出取决于介质pH的不同特征波长的发射。 这允许几种测量与指示剂接触的介质的pH的方法，包括用一次激发测量两个发射，独立地选择性地离开酸和碱形式，并测量其在单个波长或双波长处的发射，或测量特征pH依赖吸收 或荧光激发光谱。 提出了使指示剂渗透到细胞膜以测量细胞内pH的方法。