Sensitivity map of laser tweezers Raman spectroscopy for single-cell analysis of colorectal cancer

Raman spectroscopy on single, living epithelial cells captured in a laser trap is shown to have diagnostic power over colorectal cancer. This new single-cell technology comprises three major components: primary culture processing of human tissue samples to produce single-cell suspensions, Raman detection on singly trapped cells, and diagnoses of the cells by artificial neural network classifications. It is compared with DNA flow cytometry for similarities and differences. Its advantages over tissue Raman spectroscopy are also discussed. In the actual construction of a diagnostic model for colorectal cancer, real patient data were taken to generate a training set of 320 Raman spectra and a test set of 80. By incorporating outlier corrections to a conventional binary neural classifier, our network accomplished significantly better predictions than logistic regressions, with sensitivity improved from 77.5% to 86.3% and specificity improved from 81.3% to 86.3% for the training set and moderate improvements for the test set. Most important, the network approach enables a sensitivity map analysis to quantitate the relevance of each Raman band to the normal-to-cancer transform at the cell level. Our technique has direct clinic applications for diagnosing cancers and basic science potential in the study of cell dynamics of carcinogenesis.

[1]  N Stone,et al.  Assessment of fiberoptic near-infrared raman spectroscopy for diagnosis of bladder and prostate cancer. , 2005, Urology.

[2]  David J. C. MacKay,et al.  The Evidence Framework Applied to Classification Networks , 1992, Neural Computation.

[3]  James P Freyer,et al.  Raman spectroscopy detects biochemical changes due to proliferation in mammalian cell cultures. , 2005, Biophysical journal.

[4]  Giovanni Volpe,et al.  The lag phase and G1 phase of a single yeast cell monitored by Raman microspectroscopy , 2006 .

[5]  S. Lane,et al.  Micro-Raman spectroscopy detects individual neoplastic and normal hematopoietic cells. , 2006, Biophysical journal.

[6]  Measurement of the transverse spatial quantum state of light at the single-photon level. , 2005, Optics letters.

[7]  Menghong Sun,et al.  Diagnosis of colorectal cancer using Raman spectroscopy of laser-trapped single living epithelial cells. , 2006, Optics letters.

[8]  Qiuxu Wei,et al.  Study of the effect of alcohol on single human red blood cells using near‐infrared laser tweezers Raman spectroscopy , 2005 .

[9]  L. K. Hansen,et al.  Melanoma diagnosis by Raman spectroscopy and neural networks: structure alterations in proteins and lipids in intact cancer tissue. , 2004, The Journal of investigative dermatology.

[10]  David J. C. MacKay,et al.  Comparison of Approximate Methods for Handling Hyperparameters , 1999, Neural Computation.

[11]  P. Gemperline,et al.  Identification of single bacterial cells in aqueous solution using confocal laser tweezers Raman spectroscopy. , 2005, Analytical chemistry.

[12]  R. Dasari,et al.  Diagnosing breast cancer by using Raman spectroscopy. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Lars Kai Hansen,et al.  Detection of skin cancer by classification of Raman spectra , 2004, IEEE Transactions on Biomedical Engineering.

[14]  Michael S. Feld,et al.  Histological Classification of Raman Spectra of Human Coronary Artery Atherosclerosis Using Principal Component Analysis , 1999 .

[15]  R. Dasari,et al.  Identifying microcalcifications in benign and malignant breast lesions by probing differences in their chemical composition using Raman spectroscopy. , 2002, Cancer research.

[16]  David J. C. MacKay,et al.  A Practical Bayesian Framework for Backpropagation Networks , 1992, Neural Computation.

[17]  H. Barr,et al.  Raman spectroscopy for identification of epithelial cancers. , 2004, Faraday discussions.

[18]  Abigail S Haka,et al.  In vivo Raman spectral pathology of human atherosclerosis and vulnerable plaque. , 2006, Journal of biomedical optics.