Chan James W, Taylor Douglas S, Zwerdling Theodore, Lane Stephen M, Ihara Ko, Huser Thomas
Lawrence Livermore National Laboratory, Livermore, California 94551, USA.
Biophys J. 2006 Jan 15;90(2):648-56. doi: 10.1529/biophysj.105.066761. Epub 2005 Oct 20.
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.
目前用于识别肿瘤细胞并将其与正常细胞区分开来的方法通常是非特异性的、缓慢的、具有生物干扰性的,或者是这些情况的组合。在此,我们表明单细胞显微拉曼光谱法避免了这些缺点,并且可基于其生物分子拉曼特征用于区分未固定的正常人淋巴细胞与转化的Jurkat和Raji淋巴细胞系。我们证明单细胞拉曼光谱为每种细胞类型提供了高度可重复的生物分子指纹。特征峰主要归因于不同的DNA和蛋白质浓度,从而能够高度可靠地(p << 0.05)区分正常淋巴细胞与转化淋巴细胞。还通过主成分分析对光谱进行比较和分析,以证明正常细胞和转化细胞形成了不同的簇,仅使用两个主成分即可定义这些簇。该方法对癌症检测的灵敏度为98.3%,97.2%的细胞被正确分类为属于正常或转化类型。这些结果证明了共聚焦显微拉曼光谱作为一种基于内在生物分子特征进行单细胞癌症检测的临床工具的潜在应用,因此无需外源荧光标记。