基于荧光显微拉曼光谱的细菌快速解耦检测方法

Bacteria fast decoupling detection based on Raman spectra and fluorescence microscopy

在细菌检测环境中往往存在多种活细菌、死菌以及非生物微颗粒,它们形态和大小相近且耦合寄居在一起,这使得普通的菌类检测方法在检测细菌的种类和含量时很难奏效。提出一种细菌快速解耦检测方法,该方法采用荧光微视技术与拉曼光谱分析技术相结合,利用荧光微视技术实现活菌与非生物微颗粒及死菌的解耦检测,并且实现活菌的定位;利用拉曼光谱技术对定位活菌进行数据采集并结合Ward聚类分析法构建各种活菌的解耦检测模型。实验证明所提出解耦方法的对活菌的解耦检测精度达到了94%以上,且检测时间仅为普通显微拉曼光谱法的1/9。适用于烧伤伤口细菌感染检测等要求检测精度高、速度快的场合。

There is a variety of live bacteria, dead bacteria and non-biological micro-particles in bacteria detection environment. They have similar shape and size, and couple together, which makes common bacteria detection meth- ods difficult to work effectively in the detection of bacteria type and content. This paper proposes a bacteria fast decoupling detection method, which integrates fluorescent microscopy and micro-Raman spectroscopy. Fluorescent microscopy technique is used to achieve the decoupling detection of live bacteria, dead bacteria and non-biological micro-particles, and also achieve the positioning of live bacteria. Raman spectroscopy technology is used to acquire the data of the live bacteria above-mentioned, and combined with ward cluster analysis to build the detection models for various live bacteria. Experiments show that the decoupling detection accuracy of the proposed method reaches above 94% , and the detection time is only one-ninth of that for normal Raman spectroscopy. This method is suitable for the occasions where high detection precision and speed are required, such as the detection of bacterial infections in burn wounds and so on.