Patent US4544546A successfully refreshed from Google Patents.
US4544546A
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Fluorescent nucleic acid stains
Assignee: Abbott Laboratories
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Filing: 1983-07-05
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Granted: 1985-10-01
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Expires: 2003-07-05
Abstract
Abstract This disclosure relates to a class of compounds which are utilized as biological stains in fluorescent microscopy. In particular, the disclosure relates to a method for detecting and identifying various structures in a biological sample which comprises treating said biological sample with a compound of the present disclosure to form a complex which emits fluorescence when irradiated with incident light. This class of compounds has been shown to be useful in detecting a variety of structures such as, for example, viruses, bacteria, yeasts, fungi, reticulocytes and cells in biological samples.
Inventors
- 👤 Chao-Huei J. Wang
- 👤 Michael E. Jolley
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 18:00
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/52; G01N33/5005; G01N33/5094; Y; Y10; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
189 words
A method of detecting nucleic acids in a biological sample which comprises contacting said biological sample with an effective amount of a dye of the formula: ##STR8## wherein n represents an integer having a value of 0 or 1; m represents an integer having a value of 1 or 2; R represents a member selected from the group consisting of lower alkyl, lower hydroxyalkyl, dialkylaminoalkyl, pyrrolidinoalkyl and morpholinoalkyl; R 1 and R 2 independently represent a member selected from the group consisting of hydrogen, lower alkyl and lower haloalkyl; R 3 represents a member selected from the group consisting of hydrogen, lower alkyl, lower alkoxy and amino; Z represents nonmetallic atoms necessary to complete a heterocyclic nucleus selected from the group consisting of a benzothiazole nucleus, indolenine nucleus, naphthothiazole nucleus, benzoselenazole nucleus, benzoxazole nucleus, quinoline nucleus, and pyridine nucleus any of which may be substituted with lower alkyl, halo, nitro, amino and dialkylamino; and X represents an anion; thereby producing a complex between the dye and the nucleic acids; irradiating the complex with light of excitation wavelength thereby causing the complex to emit fluorescence; and measuring the emitted fluorescence.
Claim 2
Independent Claim
189 words
A method of detecting nucleic acids in a biological sample which comprises contacting said biological sample with an effective amount of a dye of the formula: ##STR8## wherein n represents an integer having a value of 0 or 1; m represents an integer having a value of 1 or 2; R represents a member selected from the group consisting of lower alkyl, lower hydroxyalkyl, dialkylaminoalkyl, pyrrolidinoalkyl and morpholinoalkyl; R 1 and R 2 independently represent a member selected from the group consisting of hydrogen, lower alkyl and lower haloalkyl; R 3 represents a member selected from the group consisting of hydrogen, lower alkyl, lower alkoxy and amino; Z represents nonmetallic atoms necessary to complete a heterocyclic nucleus selected from the group consisting of a benzothiazole nucleus, indolenine nucleus, naphthothiazole nucleus, benzoselenazole nucleus, benzoxazole nucleus, quinoline nucleus, and pyridine nucleus any of which may be substituted with lower alkyl, halo, nitro, amino and dialkylamino; and X represents an anion; thereby producing a complex between the dye and the nucleic acids; irradiating the complex with light of excitation wavelength thereby causing the complex to emit fluorescence; and measuring the emitted fluorescence.
Claim 3
Dependent Claim
64 words
A method according to claim 1 wherein n is 0; R is dialkylaminoalkyl and Z represents nonmetallic atoms necessary to complete a heterocyclic nucleus selected from the group consisting of a benzothiazole nucleus, a naphthothiazole nucleus, a benzoselenazole nucleus, a benzoxazole nucleus and substituted benzothiazole, naphthothiazole, benzoselenazole, benzoxazole nucleus wherein the substituent is selected from the class consisting of lower alkyl, halo and amino.
Claim 4
Dependent Claim
12 words
A method according to claim 2 wherein the nucleic acid is DNA.
Claim 5
Dependent Claim
69 words
A method according to claim 3 wherein m is 1; R 1 and R 2 are lower alkyl, R 3 is hydrogen and Z represents nonmetallic atoms necessary to complete a heterocyclic nucleus selected from the group consisting of benzothiazole nucleus, a naphthothiazole nucleus, a benzoselenazole nucleus and substituted benzothiazole, naphthothiazole and benzoselenazole nucleus wherein the substitutent is selected from the class consisting of lower alkyl, halo and amino.
Claim 6
Dependent Claim
19 words
A method according to claim 4 wherein X is selected from the group consisting of halogen, methylsulfate and perchlorate.
Claim 7
Dependent Claim
12 words
A method according to claim 5 wherein the dye is 3-γ-dimethylaminopropyl-2-p-dimethylaminostyrylbenzothiazolium iodide.
Claim 8
Dependent Claim
12 words
A method according to claim 5 wherein the dye is 3-γ-dimethylaminopropyl-5-chloro-2-dimethylaminostyrylbenzothiazolium iodide.
Claim 9
Dependent Claim
12 words
A method according to claim 5 wherein the dye is 3-γ-dimethylaminopropyl-2-p-dimethylaminostyryl-α-naphthothiazolium iodide.
Claim 10
Dependent Claim
12 words
A method according to claim 5 wherein the dye is 3-γ-dimethylaminopropyl-2-p-dimethylaminostyrylbenzoselenazolium iodide.
Claim 11
Dependent Claim
12 words
A method according to claim 5 wherein the dye is 3-γ-dimethylaminopropyl-5-methyl-2-p-dimethylaminostyrylbenzothiazolium iodide.
Claim 12
Independent Claim
135 words
A method of staining tissues and fixed cells in a biological sample which comprises contacting said biological sample with an effective amount of a dye of the formula: ##STR9## wherein n represents an integer having a value of 0 or 1; m represents an integer having a value of 1 or 2; R 1 and R 2 independently represent a member selected from the group consisting of hydrogen, lower alkyl and lower halo alkyl; R 3 represents a member selected from the group consisting of hydrogen, lower alkyl, lower alkoxy and amino; Z represents nonmetallic atoms necessary to complete a pyridine nucleus; R represents dialkylaminoalkyl; and X represents an anion, thereby producing a dye complex, irradiating the complex with light of excitation wavelength thereby causing the complex to emit fluorescence; and measuring the emitted fluorescence.
Claim 13
Independent Claim
135 words
A method of staining tissues and fixed cells in a biological sample which comprises contacting said biological sample with an effective amount of a dye of the formula: ##STR9## wherein n represents an integer having a value of 0 or 1; m represents an integer having a value of 1 or 2; R 1 and R 2 independently represent a member selected from the group consisting of hydrogen, lower alkyl and lower halo alkyl; R 3 represents a member selected from the group consisting of hydrogen, lower alkyl, lower alkoxy and amino; Z represents nonmetallic atoms necessary to complete a pyridine nucleus; R represents dialkylaminoalkyl; and X represents an anion, thereby producing a dye complex, irradiating the complex with light of excitation wavelength thereby causing the complex to emit fluorescence; and measuring the emitted fluorescence.
Claim 14
Dependent Claim
23 words
A method according to claim 11 wherein m is 1, R 1 and R 2 are lower alkyl and R 3 is hydrogen.
Claim 15
Dependent Claim
10 words
A method according to claim 12 wherein n is 1.
Claim 16
Dependent Claim
12 words
A method according to claim 13 wherein the dye is 1-γ-dimethylaminopropyl-4-p-dimethylaminostyrylpyridinium iodide.
Claim 17
Independent Claim
134 words
A method of staining cells in suspension in a biological sample which comprises contacting said biological sample with an effective amount of a dye of the formula: ##STR10## wherein n represents an integer having a value of 0 or 1; m represents an integer having a value of 1 or 2; R 1 and R 2 independently represent a member selected from the group consisting of hydrogen, lower alkyl, and lower haloalkyl; R 3 represents a member selected from the group consisting of hydrogen, lower alkyl, lower alkoxy and amino; Z represents nonmetallic atoms necessary to complete an aminopyridine nucleus; R represents lower alkyl; and X represents an anion; thereby producing a dye complex, irradiating the complex with light of excitation wavelength thereby causing the complex to emit fluorescence; and measuring the emitted fluorescence.
Claim 18
Independent Claim
134 words
A method of staining cells in suspension in a biological sample which comprises contacting said biological sample with an effective amount of a dye of the formula: ##STR10## wherein n represents an integer having a value of 0 or 1; m represents an integer having a value of 1 or 2; R 1 and R 2 independently represent a member selected from the group consisting of hydrogen, lower alkyl, and lower haloalkyl; R 3 represents a member selected from the group consisting of hydrogen, lower alkyl, lower alkoxy and amino; Z represents nonmetallic atoms necessary to complete an aminopyridine nucleus; R represents lower alkyl; and X represents an anion; thereby producing a dye complex, irradiating the complex with light of excitation wavelength thereby causing the complex to emit fluorescence; and measuring the emitted fluorescence.
Claim 19
Dependent Claim
23 words
A method according to claim 15 wherein m is 1, R 1 and R 2 are lower alkyl and R 3 is hydrogen.
Claim 20
Dependent Claim
10 words
A method according to claim 16 wherein n is 1.
Claim 21
Dependent Claim
12 words
A method according to claim 17 wherein the dye is 1-methyl-2-amino-4-p-dimethylaminostyrylpyridinium iodide.
References Cited (Backward Citations)
US3833863A
Cited
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1988-09-30 | 1989-04-01 | 1989-10-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-09-30 | 1993-04-01 | 1993-10-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-09-30 | 1997-04-01 | 1997-10-01 | $7700.00 | Window Closed / Lapsed |