Martin Miguela, Perez-Guaita David, Andrew Dean W, Richards Jack S, Wood Bayden R, Heraud Philip
Centre for Biospectroscopy, Monash University.
Centre for Biomedical Research, Burnet Institute.
J Vis Exp. 2018 Nov 2(141). doi: 10.3791/56797.
We demonstrate a method of quantification and detection of parasites in aqueous red blood cells (RBCs) by using a simple benchtop Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR) spectrometer in conjunction with Multivariate Data Analysis (MVDA). 3D7 P. falciparum were cultured to 10% parasitemia ring stage parasites and used to spike fresh donor isolated RBCs to create a dilution series between 0-1%. 10 µL of each sample were placed onto the center of the ATR diamond window to acquire the spectrum. The sample data was treated to improve the signal to noise ratio and to remove the contribution of water, and then the second derivative was applied to resolve spectral features. The data were then analyzed using two types of MVDA: first Principal Component Analysis (PCA) to determine any outliers and then Partial Least Squares Regression (PLS-R) to build the quantification model.
我们展示了一种通过使用简单的台式衰减全反射傅里叶变换红外（ATR-FTIR）光谱仪结合多变量数据分析（MVDA）来定量和检测水性红细胞（RBC）中寄生虫的方法。将3D7恶性疟原虫培养至10%的寄生虫血症环状期寄生虫，并用于加标新鲜供体分离的红细胞，以创建0-1%之间的稀释系列。将每个样品的10µL置于ATR金刚石窗口中心以获取光谱。对样品数据进行处理以提高信噪比并去除水的贡献，然后应用二阶导数来解析光谱特征。然后使用两种类型的MVDA对数据进行分析：首先是主成分分析（PCA）以确定任何异常值，然后是偏最小二乘回归（PLS-R）以建立定量模型。
