水体细菌微生物多波长透射光谱快速准确获取方法研究

为实现水体细菌微生物快速在线监测,搭建了多波长透射光谱快速测量实验系统,利用该系统分别测量了重铬酸钾标准溶液紫外波段及中性滤光片可见波段的透射光谱,并与紫外-可见分光光度计测得的透射光谱进行对比分析,验证了实验系统测量透射光谱的准确性;以水体中常见的金黄色葡萄球菌作为研究对象,利用搭建的实验系统获取金黄色葡萄球菌溶液在220~900nm波段的前向小角度透射光谱,进一步验证了实验系统测量细菌微生物透射光谱的准确性和快速性。结果表明,由实验系统和紫外-可见分光光度计测得的重铬酸钾标准溶液,与中性滤光片紫外波段及可见波段透射光谱的线性拟合相关系数分别为0.999 7和0.999 5,光密度误差分别在5.00%和4.58%以内,说明两个系统测量光谱的一致性较好,所搭建的实验系统测量标准样品紫外-可见透射光谱准确度较高;对于金黄色葡萄球菌,实验系统测得的透射光谱经过校正后,与紫外-可见分光光度计测得的透射光谱线性拟合的相关系数为0.999 97,两者相比的光密度误差在0.74%以内;系统重复30次细菌光谱信号采集获得平均透射光谱单次测量时间为15s,说明该实验系统相对于紫外-可见分光光度计能够快速准确获取水体细菌微生物多波长透射光谱,在保证测量结果准确的同时缩短了光谱测量时间,为水体细菌微生物快速检测提供技术支持。

Study on Rapid and Accurate Acquisition Method of Multi-Wavelength Transmission Spectroscopy of Bacteria in Water

Based on the needs of rapid on-line monitoring of bacteria microorganism in water,a multi-wavelength transmission spectrum measurement system was set up.Ultraviolet transmission spectra of potassium dichromate standard solution and visible transmission spectra of neutral filter were measured by the experiment system,which were then compared with the transmission spectra measured by ultraviolet and visible spectrophotometer to validate the accuracy of the transmission spectra measured by the experiment system.Staphylococcus aureus which commonly existed in water was studied in this paper.Moreover,small an-gle transmission spectra of staphylococcus aureus were measured over a broad range of transmission wavelength (220~900 nm) by the experiment system.The results confirmed the accuracy and rapidity of the experiment system measuring the transmission spectra of bacteria microorganism.The transmission spectra of the potassium dichromate standard solution or neutral filter meas-ured by the experiment system was strongly correlated with the transmission spectra measured by Ultraviolet and visible spectro-photometer,and their linear fitting coefficients were 0.9997 and 0.9995 (R2=0.9997 and R2=0.9995).The errors of optical densities measured by the experiment system were less than 5.00% and 4.58%,which showed that spectral consistency was better and ultraviolet and visible transmission spectra of standard samples measured by the experiment system had a high accura-cy.The correlation coefficient of transmission spectra of the staphylococcus aureus measured by the experiment system after cali-bration fitted with that of ultraviolet and visible spectrophotometer is 0.99997.As well,the error was less than 0.74% between the experimental system and ultraviolet and visible spectrophotometer.In addition,single measurement time of the average transmission spectrum measured by the system was 15s,which was 30 times of the signal acquisition.It showes that the experi-mental system can quickly and accurately obtain the multi-wavelength transmission spectra of bacterial microbial in water with re-spect to ultraviolet-visible spectrophotometer.The transmission spectrum measured by the experimental system is not only high-ly accurate,but also shortens the spectrum measurement time,which provides a technical support for the rapid detection of bac-terial microbial in water.