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

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.

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.

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.

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.