基于反射式成像光路的全波段蛋白检测仪的研制

Development of a full-band protein detection system based on reflective imaging

为了实现酶标仪的全波段和快速检测,设计出一种基于反射式成像光路的全波段蛋白检测仪,采用反射式光学检测的方式进行成像,将一维成像光程扩展为二维,提高维度减少空间,同时提高了视场范围。分别进行了Human Pentraxin 3/TSG-14(PTX3)酶联免疫吸附实验以及二喹啉甲酸法(BCA)蛋白浓度检测实验,检测波长分别为450 nm和562 nm,并完成了对生物显色实验溶液的图像采集,利用图像处理比色算法进行分析处理,完成了蛋白浓度的测定。PTX3实验和BCA实验均基于四参数拟合模型绘制拟合曲线,拟合优度分别为0.9996和0.9997,与商用酶标仪绘制的标准曲线基本一致。PTX3实验使用42份未知样本,BCA实验使用40份未知样本,基于图像处理比色算法的质量浓度测量结果与商用酶标仪检测的质量浓度误差在5%以内,验证了全波段蛋白检测仪的可行性。

In order to realize the full band and rapid detection of microplate reader,a full-band protein detection system based on reflective imaging was designed.The imaging was carried out by reflective optical detection method.The one-dimensional imaging optical path was extended to two-dimensional.The space was reduced,and the field of view was increased.Human Pentraxin 3/TSG-14(PTX3)enzyme-linked immunosorbent assay and bicinchoninic acid assay method(BCA)protein concentration assay were carried out respectively.The detection wavelength was 450 nm and 562 nm,respectively.The image collection was completed,and the colorimetric algorithm was used for analysis and processing.The determination of protein concentration was completed.The PTX3 and BCA experiments were both based on the four-parameter fitting model to draw the fitting curve,and the goodness of fit was 0.9996 and 0.9997,respectively,which was basically consistent with the standard curve drawn by commercial enzyme labelling instrument.The PTX3 experiment used 42 unknown samples,and the BCA experiment used 40 unknown samples.The concentration measurement results based on the colorimetric algorithm and the concentration detected by the commercial enzyme plate analyzer were less than 5%,which verified the feasibility and stability of the full-band protein detection system.