As shown in Figure 2, the average EFs based on the neat benzene thiol are dependent on the choice of Raman mode strongly. However, the relative Raman enhancement between our SERS substrates (including Klarite® substrate) was found to be relatively independent on the choice

of Raman mode used for comparison. For comparison, the three Raman modes associated with vibrations about the aromatic ring are presented in Figure 2c. So, to get an accurate and comparable estimation of the average enhancement factor, Raman mode used for the calculation of the average EF must be selected carefully. Here, the intensities of the peak found at 998 cm-1, carbon-hydrogen wagging mode which is the furthest mode removed from the gold surface were used to compute the average EFs [8, 42]. In addition, the average EF of Klarite® Fosbretabulin substrate was calculated to be 5.2 × 106, which is reasonable selleck chemical because the enhancement factor for the inverted pyramid structure of Klarite® substrates relative to a non-enhancing surface is rated to a lower bound of approximately 106[42]. Results and discussion The average peak intensity at 998 cm-1, the number of molecules contributing to the Raman signal, the calculated average EFs, and the relative

standard deviation (RSD) for all SERS substrates are presented in Table 1. For each substrate, more than 80 spectra were 5-Fluoracil collected at various positions to ensure that a reproducible SERS response was attained. Spatial mapping with an area larger than 20 μm × 20 μm of the SERS intensity of W-AAO2-Au was shown in Figure 2d as an example. Table 1 SERS performance parameters of SERS substrates Sample Peak intensity (counts/mW/s) Number of molecules Average EF RSD (%) P-AAO-Au 351.62 1.58 × 108 1.65 × 105 8.02 W-AAO1-Au 997.92 2.88 × 107 Epothilone B (EPO906, Patupilone) 2.56 × 106 8.25 W-AAO2-Au 1295.04 1.62 × 107 5.93 × 106 6.43 Klarite® 772.58 1.10 × 107 5.21 × 106 7.12

The average peak intensity at 998 cm-1, the calculated number of molecules, the average EFs and the RSD for P-AAO-Au, W-AAO1-Au, W-AAO2-Au, and Klarite® SERS substrates. As shown in Figure 2a,b,c and Table 1, an obvious enhancement of Raman signal of the nanowire network AAO SERS substrates (W-AAO1-Au and W-AAO2-Au) is found, compared to that of porous AAO SERS substrate (P-AAO-Au). The Raman signal of W-AAO2-Au is the strongest in all of the SERS substrates (including the Klarite® substrate). Table 1 also shows a tremendous increase of average EF of the nanowire network AAO SERS substrate comparing with porous AAO SERS substrate. The average EFs of W-AAO1-Au and W-AAO2-Au are 2.56 × 106 and 5.93 × 106, about 14 and 35 times larger than that of P-AAO-Au (1.56 × 105), respectively. Moreover, the average EF of our best SERS substrate, W-AAO2-Au, is larger than that of commercial Klarite® substrate by about 14%.