EP0445434A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Thionin stain technique
Assignee: Cytyc Corp
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Filing: 1990-12-27
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Granted: —
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Expires: 2010-12-27
Abstract
Abstract Cellular analysis in accordance with the invention involves staining cellular material with a thionin stain to distinguish the nuclear portion from the cytoplasm, counterstaining, illuminating the thionin-stained cellular material with energy in the infrared spectrum, and imaging the stained cellular material. The analysis can be automated or manual.
Inventors
- 👤 David J. Zahniser
- 👤 Petrus S. Oud
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 17:08
CPC Classifications
G; G01; G01N; G01N1/00; G01N1/28; G01N1/30; Y; Y10; Y10T; Y10T436/00; Y10T436/25
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
107 words
A method of staining cellular material for contrasting the nucleus from cytoplasmic material, for producing stained cellular material suitable for automated machine analysis, said method comprising the successive steps of staining the cellular material with a thionin stain for attaining at least a selected infrared absorbance by nuclear material, counterstaining the cellular material with a stain having an infrared absorbance by cytoplasmic material distinguishable from said infrared absorbance of nuclear material with said thionin stain, illuminating the stained cellular material with energy in the infrared spectrum, and imaging the stained cellular material in response to output energy from the stained cellular material due to said infrared illumination.
Claim 2
Dependent Claim
38 words
The method of claim 1 wherein said imaging step comprises digitizing said output energy, said imaging step further comprising the step of pathologically testing said digitized image information, and wherein said testing step comprises classifying said cellular material.
Claim 3
Dependent Claim
27 words
The method of claim 1 wherein said imaging step includes detecting said output energy with an opto-electric transducer including photomultipliers, solid-state photo-detectors, detector arrays, and television cameras.
Claim 4
Dependent Claim
31 words
The method of claim 1 wherein said counterstaining step involves staining with a counterstain compound to produce stained cellular material wherein the cytoplasmic material is relatively transparent to said infrared illumination.
Claim 5
Dependent Claim
27 words
The method of claim 1 wherein said staining step includes preparing a thionin stain consisting essentially of a thionin salt, an organic solvent, and a pH buffer.
Claim 6
Dependent Claim
20 words
The method of claim 5 wherein said organic solvent is selected from the group consisting of low molecular weight alcohols.
Claim 7
Dependent Claim
23 words
The method of claim 6 wherein said low molecular weight alcohol is selected from the group consisting of methanol, ethanol and isopropyl alcohol.
Claim 8
Dependent Claim
18 words
The method of claim 1 wherein said infrared illuminating step comprises providing energy in the near infrared spectrum.
Claim 9
Dependent Claim
111 words
The method of claim 1 wherein said counterstaining step comprises the steps of preparing a Papanicolaou-type counterstain consisting essentially of a first staining solution, a second staining solution, and alcoholic rinses, wherein said first staining solution consists essentially of an orange dye, an alcoholic solvent, and pH buffer, and wherein said second staining solution consists essentially of a green dye, a red dye, a pH buffer, and an alcoholic solvent, staining the cellular material, subsequent to said thionin-staining step, with said Papanicolaou-type counterstain, illuminating the stained cellular material with energy in the infrared spectrum, and imaging the output radiant energy from said stained cellular material in response to said infrared illumination.
Claim 10
Dependent Claim
71 words
The method of claim 1 wherein said counterstaining step comprises the steps of preparing an eosin-type counterstain consisting essentially of an Eosin Y dye, an organic solvent, and a pH buffer, staining the cellular material, subsequent to said thionin-staining step, with said eosin-type counterstain, illuminating the stained cellular material with energy in the infrared spectrum, imaging the output radiant energy from said stained cellular material in response to said infrared illumination.
References Cited (Backward Citations)
No citation records indexed.
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