Method of reconstituting cellular spectra useful for detecting cellular disorders
Embodiments of the present disclosure provides improved methods for determining the presence of abnormalities in exfoliated cells. In one embodiment, the present disclosure provides methods for reconstructing cellular spectrum of a cell sample by creating a spectral map of the cellular sample, generating a binary mask of the spectral map, removing edge artifacts from each cell, and co-adding spectral data of each pixel corresponding to the cell to reconstruct the spectrum of each cell.
1. A method of generating a spectrum of a cell, the method comprising:
(a) receiving a plurality of spectral pixels, each of the spectral pixels corresponding to a portion of the cell, each of the spectral pixels being associated with a plurality of measurements, each of the measurements being associated with an intensity of light at a particular wavenumber, one of the measurements associated with each spectral pixel being a sorting measurement, the sorting measurement being associated with a wavenumber within a band of wavenumbers;
(b) identifying a subset of the plurality of the spectral pixels, a first pixel being in the subset, the sorting measurement of the first pixel being greater than or equal to the sorting measurements of the other spectral pixels, other pixels in the plurality of spectral pixels being in the subset if they satisfy a first criteria, a spectral pixel satisfying the first criteria if that spectral pixel's sorting measurement is greater than a first threshold;
(c) generating a spectrum, the spectrum having a plurality of reconstructed measurements, each of the reconstructed measurements corresponding to a particular wavenumber, each of the reconstructed measurements being formed according to a sum of the measurements associated with a particular wavenumber of all the pixels in the subset.
2. The method according to claim 1 , the first threshold being a preselected percentage of the first pixel's sorting measurement.
3. The method according to claim 2 , the band of wavenumbers having a lower end and an upper end, the lower and upper ends being user selectable values.
4. The method according to claim 3 , the lower end being 1640 cm −1 and the upper end being 1670 cm −1 .
5. The method according to claim 3 , the sorting measurement for each spectral pixel being a peak value of that spectral pixel's associated measurements, the peak value being a peak that is closest to a user selectable wavenumber.
6. The method according to claim 5 , the user selectable wavenumber being 1650 cm −1 .
7. The method according to claim 5 , pixels in the plurality of spectral pixels being in the subset only if they satisfy both the first criteria and a second criteria, a pixel satisfying the second criteria if a difference between the wavenumber associated with the second pixel's sorting measurement and the wavenumber associated with the first pixel's sorting measurement is less than a second threshold.
8. The method according to claim 7 , the second threshold being a user selectable number.
9. The method according to claim 8 , the user selectable number being 4 cm −1 .
10. The method according to claim 1 , the measurements associated with each spectral pixel representing values derived from light intensity measurements.
11. A method of analyzing the physiological state of a test cell, the method comprising:
(a) generating a spectrum of the test cell, the method comprising:
(i) receiving a plurality of spectral pixels, each of the spectral pixels corresponding to a portion of the cell, each of the spectral pixels being associated with a plurality of measurements, each of the measurements being associated with an intensity of light at a particular wavenumber, one of the measurements associated with each spectral pixel being a sorting measurement, the sorting measurement being associated with a wavenumber within a band of wavenumbers;
(ii) identifying a subset of the plurality of the spectral pixels, a first pixel being in the subset, the sorting measurement of the first pixel being greater than or equal to the sorting measurements of the other spectral pixels, other pixels in the plurality of spectral pixels being in the subset if they satisfy a first criteria, a spectral pixel satisfying the first criteria if that spectral pixel's sorting measurement is greater than a first threshold;
(iii) generating the spectrum, the spectrum having a plurality of reconstructed measurements, each of the reconstructed measurements corresponding to a particular wavenumber, each of the reconstructed measurements being formed according to a sum of the measurements associated with a particular wavenumber of all the pixels in the subset; and
(b) determining whether the reconstructed spectrum of the test call has a predetermined criterion, the predetermined criterion being indicative of the physiological state of the test cell.
12. The method of claim 11 , wherein the predetermined criterion is generated from abnormal control epithelial cell spectra or from normal control epithelial cell spectra.
13. The method of claim 12 , wherein the epithelial cells in the test and control samples are endothelial, mesothelial or urothelial cells.
14. A method of detecting an epithelial cell disorder in a test cell, the method comprising:
(a) generating a spectrum of a test cell, the method comprising:
(i) receiving a plurality of spectral pixels, each of the spectral pixels corresponding to a portion of the cell, each of the spectral pixels being associated with a plurality of measurements, each of the measurements being associated with an intensity of light at a particular wavenumber, one of the measurements associated with each spectral pixel being a sorting measurement, the sorting measurement being associated with a wavenumber within a band of wavenumbers;
(ii) identifying a subset of the plurality of the spectral pixels, a first pixel being in the subset, the sorting measurement of the first pixel being greater than or equal to the sorting measurements of the other spectral pixels, other pixels in the plurality of spectral pixels being in the subset if they satisfy a first criteria, a spectral pixel satisfying the first criteria if that spectral pixel's sorting measurement is greater than a first threshold;
(iii) generating the spectrum, the spectrum having a plurality of reconstructed measurements, each of the reconstructed measurements corresponding to a particular wavenumber, each of the reconstructed measurements being formed according to a sum of the measurements associated with a particular wavenumber of all the pixels in the subset; and
(b) determining whether the generated spectrum of the test cell has a predetermined criterion, the predetermined criterion being indicative of the presence of a disorder in the test cell.
15. The method of claim 14 , wherein the predetermined criterion is generated from abnormal control epithelial cell spectra.
16. The method of claim 14 , wherein the epithelial cells in the test and control samples are endothelial, mesothelial or urothelial cells.
17. The method of claim 14 , wherein the epithelial cell disorder is a benign disorder, a viral disorder, pre-cancer or cancer.
18. A method analyzing a cell in a sample, the method comprising:
(a) generating a spectrum of a test cell, the method comprising:
(i) receiving a plurality of spectral pixels, each of the spectral pixels corresponding to a portion of the cell, each of the spectral pixels being associated with a plurality of measurements, each of the measurements being associated with an intensity of light at a particular wavenumber, one of the measurements associated with each spectral pixel being a sorting measurement, the sorting measurement being associated with a wavenumber within a band of wavenumbers;
(ii) identifying a subset of the plurality of the spectral pixels, a first pixel being in the subset, the sorting measurement of the first pixel being greater than or equal to the sorting measurements of the other spectral pixels, other pixels in the plurality of spectral pixels being in the subset if they satisfy a first criteria, a spectral pixel satisfying the first criteria if that spectral pixel's sorting measurement is greater than a first threshold;
(iii) generating the spectrum, the spectrum having a plurality of reconstructed measurements, each of the reconstructed measurements corresponding to a particular wavenumber, each of the reconstructed measurements being formed according to a sum of the measurements associated with a particular wavenumber of all the pixels in the subset;
(b) determining whether the generated spectrum of the test cell has a predetermined criterion, the predetermined criterion being indicative of the presence of a disorder in the test cell; and
(c) forming a reconstructed cellular spectrum, the reconstructed cellular spectrum having a plurality of reconstructed intensities, each of the reconstructed intensities corresponding to a particular wavenumber, each of the reconstructed intensities being formed according to a sum of the intensity measurements at a particular wavenumber of the pixels in the subset.