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US5635402A Expired / Lapsed US ⏱️ 0.0 years remaining

Technique for determining whether a cell is malignant as opposed to non-malignant using extrinsic fluorescence spectroscopy

Assignee: Individual
•
Filing: 1994-04-29
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Granted: 1997-06-03
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Expires: 2014-04-29
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Abstract

Abstract A technique for determining whether a cell is malignant as opposed to non-malignant using extrinsic fluorescence spectroscopy. The technique is premised on the principle that certain fluorescent dyes preferentially stain malignant cells as opposed to non-malignant cells. Accordingly, by exposing a cell to the fluorescent dye, irradiating the cell with light of such a wavelength as to cause the dye to fluoresce, measuring the intensity of fluorescence at a wavelength indicative of fluorescence of the dye, and comparing the fluorescence intensity to standards obtained from malignant cells and non-malignant cells, it is possible for one to accurately classify the cell as being either malignant or non-malignant. The present invention also relates to an automated system which applies the principles of the aforementioned technique to depict the spatial distribution of cells within an area of a Pap smear-type sample and to characterize each of the cells as being malignant or non-malignant.

Inventors

  • 👤 Robert R. Alfano
  • 👤 Cheng H. Liu
  • 👤 Wei L. Sha
  • 👤 Yury Budansky

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-26 13:27
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N21/00; G01N21/62; G01N21/63; G01N21/64; G01N21/6428; G01N33/52; G01N2021/6417; G01N2021/6421; G01N2021/6423; G01N21/645; G01N2021/6484

Claims Analysis

Total Claims: 30
Claim 1
Independent Claim 206 words
A method of examining the cells present within an area of a Pap smear for purposes of detecting cancer, the method comprising the steps of: a) exposing the cells present within the area of the Pap smear to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) illuminating the area with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; c) measuring the intensity of fluorescent light emitted from the area as a function of location within the area at a wavelength indicative of fluorescence of the fluorescent dye, said measuring step comprising passing the fluorescent light through a filter selective for said wavelength indicative of fluorescence of said fluorescent dye and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; d) comparing the electrical signals obtained in step (c) to appropriate standards obtained from cells known to be malignant and non-malignant; and e) using the results of said comparisons to generate a map of the area, said map conveying the spatial distribution of cells within the area and a characterization of each cell within the area as being either malignant or non-malignant.
Claim 2
Independent Claim 206 words
A method of examining the cells present within an area of a Pap smear for purposes of detecting cancer, the method comprising the steps of: a) exposing the cells present within the area of the Pap smear to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) illuminating the area with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; c) measuring the intensity of fluorescent light emitted from the area as a function of location within the area at a wavelength indicative of fluorescence of the fluorescent dye, said measuring step comprising passing the fluorescent light through a filter selective for said wavelength indicative of fluorescence of said fluorescent dye and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; d) comparing the electrical signals obtained in step (c) to appropriate standards obtained from cells known to be malignant and non-malignant; and e) using the results of said comparisons to generate a map of the area, said map conveying the spatial distribution of cells within the area and a characterization of each cell within the area as being either malignant or non-malignant.
Claim 3
Dependent Claim 27 words
The method as claimed in claim 1 wherein said fluorescent dye is selected from the group consisting of hematoporphyrin, hematoporphyrin derivative, dihematoporphyrin ether/ester, tetramicin, rhodamine, and cyanine.
Claim 4
Dependent Claim 14 words
The method as claimed in claim 2 wherein said fluorescent dye is hematoporphyrin derivative.
Claim 5
Dependent Claim 23 words
The method as claimed in claim 2 wherein said fluorescent dye is selected from the group consisting of hematoporphyrin, hematoporphyrin derivative, dihematoporphyrin ether/ester.
Claim 6
Dependent Claim 39 words
The method as claimed in claim 4 wherein the fluorescent dye used has an absorption spectrum and wherein said illuminating step comprises illuminating the area with light at a wavelength within the absorption spectrum for the fluorescent dye used.
Claim 7
Dependent Claim 35 words
The method as claimed in claim 5 wherein said fluorescent dye is hematoporphyrin and wherein said measuring step comprises measuring the intensity of fluorescence at a wavelength of about 820 nm or about 860 nm.
Claim 8
Dependent Claim 28 words
The method as claimed in claim 5 wherein said illuminating step comprises illuminating the area with light having a wavelength between about 300 nm and about 400 nm.
Claim 9
Dependent Claim 28 words
The method as claimed in claim 7 wherein said illuminating step comprises illuminating the area with light having a wavelength between about 350 nm and about 400 nm.
Claim 10
Dependent Claim 39 words
The method as claimed in claim 5 wherein said fluorescent dye is hematoporphyrin derivative or dihematoporphyrin ether/ester and wherein said measuring step comprises measuring the intensity of fluorescence at a wavelength of about 630 nm or about 690 nm.
Claim 11
Dependent Claim 32 words
The method as claimed in claim 9 wherein said fluorescent dye is hematoporphyrin derivative and wherein said measuring step comprises measuring the intensity of fluorescence at a wavelength of about 630 nm.
Claim 12
Dependent Claim 31 words
The method as claimed in claim 10 wherein said exposing step comprises adding an appropriate amount of hematoporphyrin derivative to the area and removing any excess hematoporphyrin derivative from the area.
Claim 13
Dependent Claim 26 words
The method as claimed in claim 11 wherein the hematoporphyrin derivative added to the cell is in solution at a concentration of approximately 1.0×10 -3 Mol/L.
Claim 14
Independent Claim 245 words
A method of examining the cells present within an area of a Pap smear for purposes of detecting cancer, the method comprising the steps of: a) exposing the cells within the area of the Pap smear to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) illuminating the area with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; c) measuring the intensity of fluorescent light emitted from the area as a function of location within the area at a reference wavelength and at a wavelength indicative of fluorescence of the fluorescent dye, said measuring step comprising passing the fluorescent light alternately through a filter selective for said wavelength indicative of fluorescence of the fluorescent dye and through a filter selective for said reference wavelength and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; d) determining the ratio of intensities of said electrical signals corresponding to the fluorescent light at said reference wavelength and at said wavelength indicative of fluorescence of the fluorescent dye; e) comparing said ratio to standard ratios obtained from malignant and non-malignant cells; and f) using the results of said comparisons to generate a map of the area, said map conveying the spatial distribution of cells within the area and a characterization of each cell within the area as being either malignant or non-malignant using said comparisons.
Claim 15
Independent Claim 245 words
A method of examining the cells present within an area of a Pap smear for purposes of detecting cancer, the method comprising the steps of: a) exposing the cells within the area of the Pap smear to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) illuminating the area with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; c) measuring the intensity of fluorescent light emitted from the area as a function of location within the area at a reference wavelength and at a wavelength indicative of fluorescence of the fluorescent dye, said measuring step comprising passing the fluorescent light alternately through a filter selective for said wavelength indicative of fluorescence of the fluorescent dye and through a filter selective for said reference wavelength and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; d) determining the ratio of intensities of said electrical signals corresponding to the fluorescent light at said reference wavelength and at said wavelength indicative of fluorescence of the fluorescent dye; e) comparing said ratio to standard ratios obtained from malignant and non-malignant cells; and f) using the results of said comparisons to generate a map of the area, said map conveying the spatial distribution of cells within the area and a characterization of each cell within the area as being either malignant or non-malignant using said comparisons.
Claim 16
Dependent Claim 27 words
The method as claimed in claim 13 wherein said fluorescent dye is selected from the group consisting of hematoporphyrin, hematoporphyrin derivative, dihematoporphyrin ether/ester, tetramicin, rhodamine, and cyanine.
Claim 17
Dependent Claim 24 words
The method as claimed in claim 13 wherein said fluorescent dye is selected from the group consisting of hematoporphyrin, hematoporphyrin derivative, and dihematoporphyrin ether/ester.
Claim 18
Dependent Claim 39 words
The method as claimed in claim 15 wherein the fluorescent dye used has an absorption spectrum and wherein said illuminating step comprises illuminating the area with light at a wavelength within the absorption spectrum for the fluorescent dye used.
Claim 19
Dependent Claim 28 words
The method as claimed in claim 16 wherein said illuminating stem comprises illuminating the area with light at a wavelength between about 300 nm and about 400 nm.
Claim 20
Dependent Claim 28 words
The method as claimed in claim 17 wherein said illuminating step comprises illuminating the area with light having a wavelength between about 350 nm and about 400 nm.
Claim 21
Dependent Claim 62 words
The method as claimed in claim 18 wherein said fluorescent dye is hematoporphyrin derivative, or dihematoporphyrin ether/ester and wherein said measuring step comprises measuring the intensity of fluorescence at a reference wavelength of about 440 nm, about 560 nm, or about 580 nm and at a wavelength indicative of fluorescence of said fluorescent dye of about 630 nm or about 690 nm.
Claim 22
Dependent Claim 59 words
The method as claimed in claim 18 wherein said fluorescent dye is hematoporphyrin and wherein said measuring step comprises measuring the intensity of fluorescence from the cell at a reference wavelength of about 440 nm about 560 nm or about 580 nm. at a wavelength at which the fluorescent dye fluoresces of about 620 nm or about 660 nm.
Claim 23
Independent Claim 214 words
A method of making a map of the spatial distribution of cells within an area of a Pap smear and for characterizing each of the cells within the area as being either malignant or non-malignant, the method comprising the steps of: a) exposing the cells within the area of the Pap smear to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) illuminating the area with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; c) measuring at a wavelength indicative of fluorescence of the fluorescent dye the fluorescence intensity of the entire illuminated area as a function of location therewithin, said measuring step comprising passing the fluorescent light through a filter selective for said wavelength indicative of fluorescence of the fluorescent dye and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; d) comparing the electrical signals emitted from each location of the entire illuminated area to standards which permit each location to be characterized as corresponding to a malignant cell, a non-malignant cell or background; e) characterizing each location on the basis of said comparisons; and f) displaying the results of said characterizations in the form of a map.
Claim 24
Independent Claim 214 words
A method of making a map of the spatial distribution of cells within an area of a Pap smear and for characterizing each of the cells within the area as being either malignant or non-malignant, the method comprising the steps of: a) exposing the cells within the area of the Pap smear to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) illuminating the area with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; c) measuring at a wavelength indicative of fluorescence of the fluorescent dye the fluorescence intensity of the entire illuminated area as a function of location therewithin, said measuring step comprising passing the fluorescent light through a filter selective for said wavelength indicative of fluorescence of the fluorescent dye and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; d) comparing the electrical signals emitted from each location of the entire illuminated area to standards which permit each location to be characterized as corresponding to a malignant cell, a non-malignant cell or background; e) characterizing each location on the basis of said comparisons; and f) displaying the results of said characterizations in the form of a map.
Claim 25
Independent Claim 234 words
A method of examining the cells present within an area of a Pap smear for purposes of detecting cancer, the method comprising the steps of: a) exposing the cells within the area of the Pap smear to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) illuminating the area with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; c) measuring the intensity of fluorescent light emitted from the area as a function of location within the area at a reference wavelength and at a wavelength indicative of fluorescence of the fluorescent dye, said measuring step comprising passing the fluorescent light alternately through a filter selective for said wavelength indicative of fluorescence of the fluorescent dye and through a filter selective for said reference wavelength and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; d) determining the difference in intensities of the electrical signals corresponding to the optical signals at said reference wavelength and at said wavelength indicative of fluorescence of the fluorescent dye; e) comparing said difference to standard differences obtained from malignant and non-malignant cells; and f) using said comparisons to generate a map conveying the spatial distribution of cells within the area and a characterization of each cell within the area as being either malignant or non-malignant.
Claim 26
Independent Claim 234 words
A method of examining the cells present within an area of a Pap smear for purposes of detecting cancer, the method comprising the steps of: a) exposing the cells within the area of the Pap smear to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) illuminating the area with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; c) measuring the intensity of fluorescent light emitted from the area as a function of location within the area at a reference wavelength and at a wavelength indicative of fluorescence of the fluorescent dye, said measuring step comprising passing the fluorescent light alternately through a filter selective for said wavelength indicative of fluorescence of the fluorescent dye and through a filter selective for said reference wavelength and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; d) determining the difference in intensities of the electrical signals corresponding to the optical signals at said reference wavelength and at said wavelength indicative of fluorescence of the fluorescent dye; e) comparing said difference to standard differences obtained from malignant and non-malignant cells; and f) using said comparisons to generate a map conveying the spatial distribution of cells within the area and a characterization of each cell within the area as being either malignant or non-malignant.
Claim 27
Independent Claim 197 words
A method of cancer-screening a cell sample containing cells whose cancer condition are unknown, the method comprising the steps of: a) exposing the cell sample to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) preparing a monolayer spread with the stained cell sample; c) illuminating an area of the monolayer spread with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; d) measuring the intensity of fluorescent light emitted from the area as a function of location within the area at a wavelength indicative of fluorescence of the fluorescent dye, said measuring step comprising passing the fluorescent light through a filter selective for said wavelength indicative of fluorescent light and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; e) comparing the electrical signals obtained in step (d) to appropriate standards obtained from cells known to be malignant and non-malignant; and f) using said comparisons to generate a map conveying the spatial distribution of cells within the area and a characterization of each cell within the area as being either malignant or non-malignant.
Claim 28
Independent Claim 197 words
A method of cancer-screening a cell sample containing cells whose cancer condition are unknown, the method comprising the steps of: a) exposing the cell sample to a fluorescent dye which preferentially stains malignant cells as opposed to non-malignant cells; b) preparing a monolayer spread with the stained cell sample; c) illuminating an area of the monolayer spread with light of a wavelength sufficient to cause the fluorescent dye to fluoresce whereby fluorescent light is emitted from the area; d) measuring the intensity of fluorescent light emitted from the area as a function of location within the area at a wavelength indicative of fluorescence of the fluorescent dye, said measuring step comprising passing the fluorescent light through a filter selective for said wavelength indicative of fluorescent light and then converting the filtered fluorescent light from optical signals into electrical signals corresponding in intensity thereto; e) comparing the electrical signals obtained in step (d) to appropriate standards obtained from cells known to be malignant and non-malignant; and f) using said comparisons to generate a map conveying the spatial distribution of cells within the area and a characterization of each cell within the area as being either malignant or non-malignant.
Claim 29
Dependent Claim 20 words
The method as claimed in claim 23 wherein said illuminating step comprises illuminating the entirety of the area at once.
Claim 30
Dependent Claim 18 words
The method as claimed in claim 23 wherein said illuminating step comprises illuminating successive portions of the area.

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 2000-06-02 2000-12-02 2001-06-03 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 2004-06-02 2004-12-02 2005-06-03 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2008-06-02 2008-12-02 2009-06-03 $7700.00 Window Closed / Lapsed

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