Possan A L, Menti C, Beltrami M, Santos A D, Roesch-Ely M, Missell F P
Centro de Ciências Exatas e Tecnologia, Universidade de Caxias do Sul, Caxias do Sul, RS, Brazil.
Instituto de Biotecnologia, Universidade de Caxias do Sul, Caxias do Sul, RS, Brazil.
Mater Sci Eng C Mater Biol Appl. 2016 Jan 1;58:541-7. doi: 10.1016/j.msec.2015.08.029. Epub 2015 Sep 3.
Escherichia coli are bacteria that must be controlled in the food industry and the hospital sector. Magnetoelastic biosensors offer the promise of rapid identification of these and other harmful antigens. In this work, strips of amorphous Metglas 2826MB3 were cut to size (5 mm × 1 mm) with a microdicing saw and were then coated with thin layers of Cr and Au, as verified by Rutherford backscattering spectroscopy (RBS). Several sensor surfaces were studied: 1) as-cast strip, wheel side; 2) as-cast strip, free surface; and 3) thinned and polished surface. A layer of cystamine was applied to the Au-covered magnetoelastic substrate, forming a self-assembledmonolayer (SAM), followed by antibodies, using a modified Hermanson protocol. The cystamine layer growth was verified by Fourier transform infrared spectroscopy (FTIR) and scanning electronmicroscopy (SEM). The biosensors were exposed to solutions of bacteria and the resonant frequency of the sensors was measured with an impedance analyzer for times up to 100 min. Reductions in the resonant frequency, corresponding to bacteria capture, were measured after optimizing the signal amplitude. For times up to 40 min, high capture rates were observed and thereafter saturation occurred. Saturation values of the frequency shifts were compared with the number of bacteria observed on the sensor using fluorescence microscopy. Parameters associated with capture kinetics were studied for different sensor surfaces. The rough surfaces were found to show a faster response, while the thinned and polished sensors showed the largest frequency shift.
大肠杆菌是食品工业和医院领域必须加以控制的细菌。磁弹性生物传感器有望快速识别这些以及其他有害抗原。在这项工作中,用微切割锯将非晶态Metglas 2826MB3条带切割成尺寸为(5毫米×1毫米),然后用卢瑟福背散射光谱法(RBS)验证后,在其表面涂覆Cr和Au薄层。研究了几种传感器表面:1)铸态条带,轮侧;2)铸态条带,自由表面;3)减薄和抛光表面。在覆盖有Au的磁弹性基底上施加一层胱胺,形成自组装单分子层(SAM),然后按照改良的赫尔曼森方案连接抗体。通过傅里叶变换红外光谱法(FTIR)和扫描电子显微镜(SEM)验证胱胺层的生长情况。将生物传感器暴露于细菌溶液中,并用阻抗分析仪测量传感器的共振频率,测量时间长达100分钟。在优化信号幅度后,测量与细菌捕获相对应的共振频率降低情况。在长达40分钟的时间内,观察到高捕获率,此后出现饱和现象。将频率偏移的饱和值与使用荧光显微镜在传感器上观察到的细菌数量进行比较。针对不同的传感器表面研究了与捕获动力学相关的参数。发现粗糙表面显示出更快的响应,而减薄和抛光的传感器显示出最大的频率偏移。