ZHANG Jingran, BI Yanrui, JING Bowen, QIAO Jian, YU Miao, SHI Guangfeng, LI Jing
The detection of myoglobin holds significant importance in the prevention of acute myocardial infarction (AMI). Myoglobin is recognized as a primary biomarker for AMI prevention. Owing to its small molecular size, myoglobin is released into the bloodstream within 1 h after the onset of chest pain and reaches peak levels within 2 h, whereas troponin and creatine kinase are released after 3 h and 6 h, respectively, thereby establishing myoglobin as a more accurate biomarker for AMI diagnosis. In recent years, surface-enhanced Raman scattering (SERS) technology has been widely adopted for biomolecule detection owing to its advantages of nondestructive analysis, high sensitivity, and rapid response. Noble metal / two-dimensional material composite SERS substrates not only exhibit high enhancement effects of noble metals but also benefit from the strong fluorescence quenching capability, high adsorption capacity, and large specific surface area of 2D materials. Additionally, three-dimensional micro/nanostructures are known to enhance SERS substrate performance, with the enhancement effects closely related to the dimensions of these structures. In this study, a composite SERS substrate comprising a molybdenum disulfide-gold-square array structure is fabricated via a combination of focused ion beam (FIB) etching, magnetron sputtering, and drop-coating methods. This substrate is designed for the label-free and highly sensitive detection of the biomolecule myoglobin, offering potential applications in biotherapy and medical diagnostics. First, a nanoarray structure is fabricated using FIB technology. The system is equipped with an electron beam imaging resolution of 0.8 nm, an ion beam imaging resolution of 4 nm, and a machining accuracy of 5 nm. A gallium ion source is employed with an acceleration voltage of 30 keV, a beam current of 24 pA, and a dwell time of 2 μs. Square array structures with varying side lengths (300, 350, and 400 nm), spacings (424, 495, and 566 nm), and depths are etched onto a clean silicon substrate. Subsequently, gold nanoparticles are deposited onto the square array structure via magnetron sputtering using a high-vacuum coating system. This process forms a gold-square array structure with localized surface plasmon resonance (LSPR) effects, significantly improving the detection resolution of the SERS substrate. The sputtering power is set to 150 W, with argon gas used as the working medium. Finally, a MoS2 solution is drop-coated onto the gold-square array structure, allowed to spread uniformly, and air-dried to form a MoS2-Au-square array composite SERS substrate. The square array structures and MoS2-Au-square array composite SERS substrate are characterized using scanning electron microscopy (SEM) and atomic force microscopy (AFM). For square arrays with side lengths of 300, 350, and 400 nm and spacings of 424, 495, and 566 nm (denoted as L1S2, L2S5, and L3S6, respectively), the overall morphology remains unchanged, exhibiting a state of adjacent contact. When the side length is fixed at 300 nm and the spacings are varied (504, 424, 344, and 264 nm), the adjacent arrays transition from separated to overlapping states. The SEM analysis of the composite substrate confirms a uniform coverage of the MoS2 film within the gaps and interior of the square arrays, verifying successful film adhesion. Raman spectroscopy and energy-dispersive X-ray spectroscopy (EDS) further validate the presence of characteristic MoS2 peaks and elemental composition, confirming the successful fabrication of the composite substrate. The Raman performance of the gold-square array and MoS2-Au-square array composite substrates is investigated using rhodamine 6G (R6G) as a probe molecule. The results indicate that the Au-L1S2 substrate (300 nm side length) exhibits stronger Raman signals compared to Au-L2S5 and Au-L3S6. Similarly, the Au-L1S2 substrate with a 424 nm spacing outperforms Au-L1S1, Au-L1S3, Au-L1S4, and Au-L1S5. The MoS2-Au-L1S2 composite substrate demonstrates the highest signal enhancement. This substrate achieves a detection limit of 10-8 mol / L for R6G, with a relative standard deviation (RSD) of 4.66%. After 7 d and 30 d of storage, the Raman intensities at 613, 1 362, and 1 650 cm-1 decrease by 9.7% and 47.6%, 5.6% and 41.8%, and 8% and 45.5%, respectively, demonstrating excellent sensitivity, uniformity, and stability. Furthermore, the composite substrate successfully detects myoglobin at a concentration of 0.02 μg / mL that is below the threshold observed in AMI cases, highlighting its potential for high-sensitivity and uniform biomarker detection.