Classification of Raman spectra of single cells with autofluorescence suppression by wavelength modulated excitation
Classification of Raman spectra of single cells with autofluorescence suppression by wavelength modulated excitation
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DOI：
10.1039/c3ay40193f
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发表时间：
2013-01-01
影响因子：
3.1
通讯作者：
Popp, Juergen
中科院分区：
文献类型：
--
作者：
Dochow, Sebastian;Bergner, Norbert;Popp, Juergen
波长调制拉曼光谱最近被证明可以抑制样品和衬底产生的荧光背景。在这里，我们应用该技术收集来自697个单个细胞的波长调制拉曼光谱，用于循环肿瘤细胞模型系统，该系统由来自患者血液的白细胞，急性髓性白血病细胞（OCI-AML3）和乳腺肿瘤细胞BT-20和MCF-7组成。我们研究了波长调制拉曼光谱的分类行为，并与化学计量学中常用的背景校正方法进行了比较。比较了基于径向核函数的支持向量机对每个细胞的经典拉曼光谱、平均拉曼光谱和波长调制拉曼光谱的分类。将数据集分为80%的训练谱和20%的独立验证谱。随机选取数据集进行200次训练和验证，检验分类的稳定性。结果显示在方框晶须图中。基于波长调制拉曼光谱的细胞识别给出了与经典和平均拉曼光谱相似的分类率，但有降低精度和增加建模变化的趋势。讨论了进一步改进波长调制拉曼光谱的可能解释和策略。
Wavelength modulated Raman spectroscopy has recently been shown to suppress the fluorescence background generated by the sample and the substrate. Here we apply this technique to collect wavelength modulated Raman spectra from 697 individual cells for a model system of circulating tumour cells that consists of leukocytes from patient's blood, acute myeloid leukaemia cells (OCI-AML3), and breast tumour cells BT-20 and MCF-7. We study the classification behaviour of wavelength modulated Raman spectra in comparison to a common background correction method in chemometrics. Classifications using a support vector machine with a radial based kernel function were compared for classical Raman spectra, average Raman spectra of each cell and wavelength modulated Raman spectra. The dataset was divided into 80% training spectra and 20% independent validation spectra. The stability of the classification was tested by performing training and validation 200 times with randomly selected datasets. The results are displayed in box whisker plots. Cell identification based on wavelength modulated Raman spectra gives similar classification rates than classical and averaged Raman spectra with a tendency of reduced accuracies and increased modelling variations. Possible explanations and strategies to further improve the wavelength modulated Raman spectroscopy are discussed.