傅里叶变换红外光谱显微技术在宫颈恶性肿瘤筛查中对细胞类型及潜在混杂变量的研究
FTIR microspectroscopic study of cell types and potential confounding variables in screening for cervical malignancies.
作者信息
Wood B R, Quinn M A, Tait B, Ashdown M, Hislop T, Romeo M, McNaughton D
机构信息
Department of Chemistry, Monash University, Clayton, Victoria, Australia.
出版信息
Biospectroscopy. 1998;4(2):75-91. doi: 10.1002/(SICI)1520-6343(1998)4:2%3C75::AID-BSPY1%3E3.0.CO;2-R.
FTIR microscopy was applied to the analysis of cell types and other variables present in Pap smears to ascertain the limitations of infrared spectroscopy in the diagnosis of cervical cancer and dysplasia. It was found that leukocytes, and in particular lymphocytes, have spectral features in the phosphodiester region (1300-900 cm[-1]) suggestive of what has previously been described as changes indicative of malignancy. Endocervical cells and fibroblasts have similar spectral features to HeLa cells and consequently could also confound diagnosis. The use of ethanol as a fixative and dehydrating agent results in retention of glycogen in cervical cell types and thus minimizes spectral changes in the glycogen region due to sampling technique. Spectra of seminal fluids exhibit strong bands in the phosphodiester/carbohydrate region; however, sperm contamination should be easily detectable by the presence of a distinctive doublet at 981/968 cm(-1). Erythrocyte spectra exhibit a reduction in glycogen band intensity, but can be discerned by a relatively low-intensity nu(s) PO2- band. Endocervical mucin spectra exhibit a reduction in glycogen bands and a very pronounced nu(s) PO2- band, which is similar in intensity to the corresponding band in HeLa cells. Thrombocytes have strong bands in the phosphodiester region, but thrombocytes can be discerned from other cell types by the presence of two small broad bands at 980 and 935 cm(-1). Candida albicans is characterized by strong bands in the polysaccharide region which could potentially obscure diagnostic bands if C. albicans is present in large numbers. Spectra of bacteria common to the female genital tract, in general, also have strong absorptions in the polysaccharide region; however, bacterial contamination is usually minimal and would not be expected to obscure cervical cell spectra. Nylon threads and bristles from cervical sampling implements produce characteristic IR profiles which allow for easy identification. Given the number of potential confounding variables associated with cervical cytology, a multivariate statistical or neural network analysis would appear to be necessary before the implementation of FTIR technology in clinical laboratories.
傅里叶变换红外光谱显微镜被用于分析巴氏涂片样本中存在的细胞类型及其他变量,以确定红外光谱在宫颈癌和发育异常诊断中的局限性。研究发现,白细胞,尤其是淋巴细胞,在磷酸二酯区域(1300 - 900 cm⁻¹)具有光谱特征,这表明之前所描述的与恶性肿瘤相关的变化。宫颈内膜细胞和成纤维细胞与海拉细胞具有相似的光谱特征,因此也可能混淆诊断。使用乙醇作为固定剂和脱水剂可使宫颈细胞类型中的糖原得以保留,从而将由于采样技术导致的糖原区域光谱变化降至最低。精液光谱在磷酸二酯/碳水化合物区域呈现强吸收带;然而,通过981/968 cm⁻¹处独特的双峰应该很容易检测到精子污染。红细胞光谱显示糖原带强度降低,但可通过相对低强度的νₛ PO₂⁻带进行辨别。宫颈内膜粘蛋白光谱显示糖原带减少以及非常明显的νₛ PO₂⁻带,其强度与海拉细胞中的相应带相似。血小板在磷酸二酯区域有强吸收带,但通过980和935 cm⁻¹处的两个小宽带可将血小板与其他细胞类型区分开来。白色念珠菌的特征是在多糖区域有强吸收带,如果大量存在白色念珠菌,可能会掩盖诊断带。女性生殖道常见细菌的光谱通常在多糖区域也有强烈吸收;然而,细菌污染通常很少,预计不会掩盖宫颈细胞光谱。宫颈采样器具上的尼龙线和刷毛会产生特征性的红外图谱,便于识别。鉴于与宫颈细胞学相关的潜在混杂变量数量众多,在临床实验室应用傅里叶变换红外光谱技术之前,似乎有必要进行多变量统计或神经网络分析。