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• W2125356973 abstract "Regular stripes of tobacco mosaic viruses (TMV) with variable line spacings (290 nm to 1 μm) are generated over large areas via printing prealigned TMVs from wrinkled poly(dimethylsiloxane) substrates onto flat substrates. The fabrication of small structures has attracted increasing interest in the past few decades. One widespread technique to achieve patterned surfaces is microcontact printing (μCP). The concept of μCP is to transfer a solution of the desired substance (ink) onto a flat surface via a structured, elastomeric stamp. It is a very flexible method as the ink and the stamp can be varied. Therefore, it is possible to produce surface patterns that differ in geometry and in the printed substance itself. Initially used for printing self-assembled monolayers (SAMs) of organic components,1 today the applications vary from small molecules to macromolecules, nanoparticles, and biological systems.2-6 While the well established μCP technique has a lot of advantages in terms of flexibility, it has one significant drawback: the elastomeric stamps are usually made by replica molding.7 Although the molding process itself is simple and cost effective, the process relies on a lithographically manufactured master.8 Therefore, it is a rather expensive method (in terms of operating costs), which reduces the accessibility for a wide range of operators. Recently, we established an alternative approach to generate stamps in a lithography-free way. The effort made use of controlled wrinkling of poly(dimethylsiloxane) (PDMS) to produce the elastomeric stamps for μCP.3 Using this technique, it was possible to generate arrays of bovine serum albumin (BSA) on a SiOx surface, where the width of the stripes is on the order of <1 μm. In particular, the alignment and assembly of biomacromolecules has attracted intense interest in recent years.9-14 As the monodispersity of these molecules is a given by nature, they form an excellent model system to study and predict the behavior of geometrically well defined systems. In this Communication, we show that it is possible to print regular virus stripes with variable line spacings without the use of any lithographic technique. The distances between the striped arrays shown here vary between ≈300 nm and ≈1 μm. Furthermore, patterns of highly aligned viruses were obtained with single-line dimensions on the order of several micrometers in length but only a few nanometers in width. This approach is based on recently published work15, 16 that showed the alignment of the well known tobacco mosaic virus (TMV)17 and silica nanoparticles on wrinkled PDMS substrates. Here, we use the wrinkled patterns as stamps to transfer the prealigned TMV onto a flat substrate (see Figure 1). The difference between conventional μCP and this method is that the ink resides in the grooves of the relief structure and not on the top.10, 18 Therefore, in the following, we will refer to this as intaglio printing (IntP). General scheme showing the various processing steps from the generation of the wrinkled substrate via stretching and plasma treatment of the PDMS block to the spin-coating of the TMV solution onto the substrate, the virus alignment in the grooves, and the final printing process. The stamp is prepared by a procedure for wrinkling elastomers previously described by various groups.19-21 Highly ordered, wrinkled topographies can be obtained by generating a hard oxide layer on an elastomeric substrate under strain. Figure 2a shows a scanning force microscopy (SFM) image of a wrinkled PDMS substrate (left), where the sinusoidal shape can be seen in the corresponding height profile (right). Typically, small pieces of PDMS (6 mm in width and 30 mm in length) were positioned in a stretching apparatus and extended from 110% to 130% of its original length. For the generation of the oxide layer, an air plasma was applied to the stretched PDMS substrate (between 20 and 75 s, 1 mbar, 18 W, PDC-32 G, Harrick). Due to the variation of the applied strain and the thickness of the oxide layer (achieved by different plasma-exposure times), the wavelength λ and the amplitude A of the wrinkles can be tuned easily.3, 19, 22 Therefore, the preparation of topographically structured substrates with variable and well controlled dimensions is possible with a straightforward and cost-effective method. These substrates will be used as stamps in the IntP method described in the following. SFM height images (left) with corresponding height profiles (right) of a) wrinkled PDMS, b) aligned TMV in wrinkled PDMS, and c) printed TMV on a on a silicon wafer, where the white lines in the SFM images represent the position of the corresponding height profile [z range = 0–50 nm for (a) and (b); z range = 0−30 nm for (c)]. The distance between the virus stripes is 288 nm. One of the key steps to generate regular TMV stripes on a flat surface is the prealignment of the virus (ink) on the stamp. The optimum conditions for the arrangement of TMV on wrinkled templates were elaborated previously.15 It is necessary for a successful IntP process that the virus is mainly located in the grooves of the wrinkles and that nearly no virus is found outside the grooves (Figure 1). An SFM image of TMV in the grooves of a PDMS stamp is shown in Figure 2b. Both the SFM image (left), but especially its cross section (right), verify that the substrate grooves are filled with the virus. To obtain those structures, 50 μL of a TMV solution with a concentration of 0.9 mg mL−1 is spin cast onto the wrinkled substrate with a spin speed of 3000 rpm. One has to keep in mind that PDMS generally has a hydrophobic surface. Therefore, a fresh oxide layer on the PDMS substrate is required; that is, the stamps should be used directly after production. The arrangement of the virus in the grooves of the wrinkles is a consequence of a dewetting process, where the virus solution forms channels in the grooves of the wrinkles.15 After spin casting, the virus solution on the PDMS substrate is dried for 30 s under a flow of nitrogen. The printing process is performed by positioning the stamp on a commercially available silicon wafer (exposed to air plasma for 1 min to hydrophilize and clean it). For the duration of 30 s, light manual pressure is applied to obtain full contact between the stamp and the SiOx surface. The stamp is then gently stripped from the surface. After drying, characterization was performed with either SFM or scanning electron microscopy (SEM). Figure 2c shows an SFM image of a TMV-patterned silicon wafer. Regular virus stripes with a distance of ≈288 nm are observed for a stamp with a wavelength λ of 285 ± 7 nm, proving that the spacing of the printed stripes is defined by the wavelength of the stamp. Not only the wavelength λ but also the amplitude A of the stamp has an influence on the resulting patterns. We successfully employed the IntP method for TMV with an amplitude range of the stamp from around 20 nm to 65 nm. With amplitudes higher than 65 nm, poor or even no particle transfer onto the wafers was observed. This upper limit in the amplitude may be explained by the thickness of the residual water film in the grooves after the spin-coating process, which is suspected to act as a mediating agent for the transfer of the TMV. Test experiments with samples dried for several days did not show any pattern transfer. As described above, the water film dewets and forms channels in the grooves of the wafer. During the printing process, these water channels wet the freshly hydrophilized SiOx surface. Thus, the contact between the wafer and the stamp is established, which leads to an effective transfer of the virus to the SiOx surface. When the amplitude of the stamp is too high, the water meniscus between the grooves of the wrinkles may be too shallow and the water does not come into contact with the silicon wafer anymore. As the amplitude range also limits the range of accessible (i.e., printable) wavelengths, we conclude that, so far, our method allows us to print stripes of wavelengths between around 290 nm to 1 μm. However, conducting the printing process in a moisturized atmosphere may overcome the described limitations. When ideal conditions are used (meaning appropriate amplitudes of the stamp and the right spin speed for the alignment process), large arrays of virus stripes are obtained. An SEM image of such a pattern is shown in Figure 3. Although some of the viruses are not in line, and some defects along the TMV strings are found, the preferential direction of the stripes with regular line spacing is clearly visible. The defects of the structure are predominantly caused by inappropriately aligned TMVs disturbing the pattern transfer. Nevertheless, on a 0.23 in2 (1 in = 2.54 cm) substrate, we found regularly aligned TMV-stripes, each several micrometers in length but only a few nanometers in width. SEM image of TMV printed on silicon wafer. The distance between the single stripes is ≈290 nm. The aim of this work is to show that the simple production of defined virus strings with variable spacings is possible with a low-cost method. As already mentioned above, the wavelength of the stamps can be modified by the plasma-exposure dose. Wrinkles with different wavelengths were generated and used as stamps for the IntP process, where the obtained TMV patterns are predefined by the topography of the wrinkles. In Figure 4, SEM and SFM images with interline spacings ranging from around 400 nm to almost 1 μm are illustrated (for a spacing of 288 nm, see Figure 3). The average wavelengths of the stamps are 404 ± 5 nm (Figure 4a), 481 ± 6 nm (Figure 4b), 631 ± 10 nm (Figure 4c), and 929 ± 13 nm (Figure 4d), respectively. As a result, virus strings with desired distances are only dependent on the choice of the stamp and the spacing of the printed virus arrays is strongly correlated to the wavelength of the wrinkles from which they are printed. In addition, this stamping process is not limited to lines as the form of the stamp dictates the geometry of the pattern. Therefore, in principle, also zig-zag structures, stars or rings, and so on may be feasible.21 Moreover, the high surface functionality of TMV opens a simple route for subsequent chemical modifications of the virus capsid via, for example, metallization or mineralization,23, 24 thus enabling the generation of metallic nanowires and arrays for electronics or optical applications. SEM images of TMV stripes with insets of SFM images (z range = 0–30 nm, scale bar = 300 nm) of the same sample (left). The cross sections of the height profiles (right) are represented by the white lines in the SFM images. The distances between the virus stripes are a) 405 nm, b) 482 nm, c) 622 nm, and d) 934 nm. In summary, we have shown that a simple intaglio printing method can be employed to obtain regular virus stripes over large areas. The interline distances of the virus patterns can be varied between 288 nm and almost 1 μm by using wrinkled PDMS stamps of different wavelengths, which are produced in a lithography-free way. By tuning the exposure time to air-plasma oxidizing the surface of a bulk PDMS sample, the wavelength of the stamp can be adjusted. Moreover, we found an amplitude limit for the stamps above which poor or no printing was achieved. In conclusion, the virus patterns produced may be used as templates for the production of metallic nanowires or serve as anchors for cell-adhesion experiments. In addition, we believe that applying the described IntP procedure could offer a low-cost method for the alignment and pattern transfer of various large molecules or nanoparticles. The authors thank Qian Wang (University of South Carolina) for providing the virus and A. Mihut, H. G. Schoberth, and C. Pester for helpful discussions. This work is supported by the Lichtenberg-Program of the VolkswagenStiftung and the Sonderforschungsbereich 481 funded by the German Science Foundation (DFG)." @default.
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• W2125356973 date "2010-09-03" @default.
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• W2125356973 title "Ordering and Printing Virus Arrays: A Straightforward Way to Functionalize Surfaces" @default.
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