**3. Results and discussion**

56 Advances in Unconventional Lithography

position, and left in contact with the substrate for at least 3 h (Figure 1c). A suspension of 65 nm particles was then added to obtain a final concentration of 0.1 %, and the system was gently stirred and left resting for 1 more hour (Figure 1d). The samples were rinsed and

Substrates with adhering 470 nm particles (Figure 1c), rinsed and dried, were again conditioned with PAH (10-5 M, pH=7, I=10-3 M) (Figure 1e), rinsed with water then with isopropanol and dried, as described for the adhesion of a single colloid layer. These samples were then placed horizontally in wells of another culture plate. 1ml of a 0.1% suspension of 65 nm particles was poured into wells and left in contact with the substrate for at least 2 h (Figure 1f). The samples were rinsed and dried using the same procedure as described for

The samples were examined by scanning electron microscopy (SEM) using a high resolution Digital Scanning microscope 982 Gemini (Leo Electron Microscopy, UK) operating at 1 or 2 kV, without any metal coating. SEM images of substrates covered with a monolayer of adhering particles were analyzed after image processing with Adobe Photoshop (version 8). In this way, the white rings defining latex particles were extracted and intensified in order to draw circles for digital processing. The images were binarized and the circles were filled before measurements (Visilog, version 6). A minimum of 500 particles was counted on more than 4 images with 10 000x magnification. The degree of particle coverage was measured as the area of particles divided by the total area. Manual counting was also performed on at least 3 images with 10 000x or 20 000x magnification of each sample. Regarding surfaces with bimodal roughness, no image processing was found suitable for extracting colloidal particles from the background. Manual counting of the 470 nm particles only was performed

The surface chemical analysis by X-ray photoelectron spectroscopy (XPS) was performed using a Kratos Axis Ultra Spectrometer (Kratos Analytical, UK) equipped with a monochromatized aluminium X-ray source (powered at 10 mA and 15 kV) and eight channeltrons detector. Charge stabilization was achieved using an electron source (filament current set at 1.9 A, bias -1.1 eV) mounted coaxial to the lens column and a charge balance plate (voltage set at -2.3 V). The rectangular analyzed area was about 0.7 mm 0.3 mm. For recording individual peaks, the pass energy was set at 40 eV. In these conditions, the full width at half maximum (FWHM) of the Ag 3d5/2 peak was about 0.9 eV. The pressure in the analysis chamber was around 10-6 Pa. The following sequence of spectra was recorded: survey spectrum, C 1s, O 1s, N 1s, Si 2p, Cl 2p, B 1s, Na 1s, Zn 2p, Ti 2p, Al 2p, S 2p and C 1s again to check the stability of charge compensation and the absence of sample degradation as a function of time. The binding energy scale was set by fixing the C 1s component due to carbon only bound to carbon and hydrogen at 284.8 eV. The data analysis was performed with the CasaXPS program (Casa Software, UK). Molar concentration ratios were calculated using peak areas normalized on the basis of the acquisition parameters, sensitivity factors, and transmission function provided by the manufacturer. Angle-resolved XPS analyses were performed by using angles between the normal to the sample surface and the direction

dried using the same procedure as described for adhesion of a single colloid layer.

**2.2.4 Surfaces with bimodal II roughness** 

adhesion of a single colloid layer.

**2.3 Surface characterization** 

on at least 4 images.

of photoelectron collection, , equal to 0°, 45° and 60°.
