Micro-Raman Spectroscopy Detects Individual Neoplastic and Normal Hematopoietic Cells

Abstract

Current methods for identifying neoplastic cells and discerning them from their normal counterparts are often nonspecific, slow, biologically perturbing, or a combination thereof. Here, we show that single-cell micro-Raman spectroscopy averts these shortcomings and can be used to discriminate between unfixed normal human lymphocytes and transformed Jurkat and Raji lymphocyte cell lines based on their biomolecular Raman signatures. We demonstrate that single-cell Raman spectra provide a highly reproducible biomolecular fingerprint of each cell type. Characteristic peaks, mostly due to different DNA and protein concentrations, allow for discerning normal lymphocytes from transformed lymphocytes with high confidence (p << 0.05). Spectra are also compared and analyzed by principal component analysis to demonstrate that normal and transformed cells form distinct clusters that can be defined using just two principal components. The method is shown to have a sensitivity of 98.3% for cancer detection, with 97.2% of the cells being correctly classified as belonging to the normal or transformed type. These results demonstrate the potential application of confocal micro-Raman spectroscopy as a clinical tool for single cancer cell detection based on intrinsic biomolecular signatures, therefore eliminating the need for exogenous fluorescent labeling.

Chapter 10 Infrared and Raman Microscopy in Cell Biology

“…8). Very similar results have been reported by Chan et al (2006) using RA-MSP, followed by PCA analysis. A largescale study of normal and cancerous urothelial cells is presently being carried out in the authors' laboratory.…”

Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells

“…Shortly thereafter it was shown, that if combined with confocal laser microscopy, Raman spectroscopy can be used to obtain information from individual living cells even with subcellular resolution [18]. Since then, the applications of Raman spectroscopy to single cells have grown exponentially [23,24], and applications related to single cancer cells [25], human embryonic stem cells [26], and even biological particles as small as lipoproteins [27] and bacterial spores [28] have been demonstrated. In addition, the field of coherent Raman microscopy has emerged, which enables label-free imaging of individual cells and tissues in vitro and in vivo [23,24,[29][30][31][32].…”

Cellular discrimination using in vitro Raman micro spectroscopy: the role of the nucleolus

“…5,6 Indeed, Raman micro spectroscopy has been shown to provide high specificity and sensitivity, even for pre-cancer detection, [7][8][9][10] and is noninvasive and potentially automatable, thus avoiding the disadvantages of many biomedical techniques used to identify and sort cancerous cells from normal ones, which exhibit low specificity and are destructive or perturb the cellular biology. [11][12][13] As an optical microscopic technique, Raman micro spectroscopy also has the potential to probe the molecular structure on a cellular and subcellular level. 14,15 In comparison to Infrared absorption spectroscopy; Raman micro spectroscopy offers the possibility to study biological matrices in an aqueous environment due to the weak contribution from water.…”