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Cavity-type DNA Origami-Based Plasmonic Nanostructures for Raman Enhancement Mengzhen Zhao, Xu Wang, Shaokang Ren, Yikang Xing, Jun Wang, Nan Teng, Dongxia Zhao, Wei Liu, Dan Zhu, Shao Su, Jiye Shi, Shiping Song, Lihua Wang, Jie Chao, and Lianhui Wang ACS Appl. Mater. Interfaces, Just Accepted Manuscript • DOI: 10.1021/acsami.7b05959 • Publication Date (Web): 16 Jun 2017 Downloaded from http://pubs.acs.org on June 22, 2017
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Cavity-type DNA Origami-based Plasmonic Nanostructures for Raman Enhancement Mengzhen Zhao, †,∥ Xu Wang,†,∥ Shaokang Ren, † Yikang Xing, † Jun Wang, † Nan Teng, † Dongxia Zhao, † Wei Liu, † Dan Zhu, † Shao Su, † Jiye Shi, § Shiping Song, ‡ Lihua Wang, ‡ Jie Chao†,* and Lianhui Wang †,* †
Key Laboratory for Organic Electronics & Information Displays (KLOEID), Institute of
Advanced Materials (IAM), National Syngerstic Innovation Center for Advanced Materials (SICAM), Nanjing University of Posts & Telecommunications, 9 Wenyuan Road, Nanjing 210023, China ‡
Division of Physical Biology &Bioimaging Center, Shanghai Synchrotron Radiation Facility,
CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China §
UCB Pharma, 208 Bath Road, Slough, SL1 3WE, UK
KEYWORDS: DNA self-assembly, Cavity-type DNA origami, Plasmonic nanostructures, Gold nanoparticles, SERS
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ABSTRACT DNA origami has been established as addressable templates for site-specific anchoring of gold nanoparticles (AuNPs). Given that AuNPs are assembled by charged DNA oligonucleotides, it is important to reduce the charge repulsion between AuNPs-DNA and the template to realize high yields. Herein, we developed a cavity-type DNA origami as templates to organize 30-nm AuNPs, which formed dimer and tetramer plasmonic nanostructures. Transmission electron microscopy (TEM) images showed that high yields of dimer and tetramer plasmonic nanostructures were obtained by using the cavity-type DNA origami as the template. More importantly, we observed significant Raman signal enhancement from molecules covalently attached to the plasmonic nanostructures, which provides a new way to highsensitivity Raman sensing.
INTRODUCTION Plasmonic nanostructures have attracted increasing attention because of their potential applications including sub-diffraction limit imaging, energy harvesting and biosensing.1-15 Compared to the top-down lithography, the bottom-up self-assembly approaches are superior in many aspects to fabricate the plasmonic nanostructures such as making three-dimensional structures and cost-effective mass production.16-18 Owing to the versatile chemistry of DNA molecules, DNA nanostructures with custom designed shapes offer new opportunities to fabricate plasmonic nanostructures with high complexity and hierarchy.19-23 Ever since Seeman pioneered the field of DNA nanotechnology in 1980s, numerous DNA nanostructures with custom designed shapes and precisely controlled motions has been designed
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and fabricated.24-26 Because of their unique self-recognition properties and high precise addressability, DNA molecules and DNA nanostructures are employed as linkers or template to self-assemble plasmonic nanostructures. Elegant examples can date back to 1996, when Mirkin and Alivisatos employed DNA and AuNPs to form macroscopic colloidal crystal lattices or separate AuNPs in a predefined distance.27,28 DNA origami technology broke through the traditional rules that employ a long, single-stranded DNA (ssDNA) scaffold with hundreds of staples to fold into arbitrary shapes in a ∼100 nm scale.29 Nowadays, DNA origami nanostructures in multiple dimensions have been established as excellent templates or linkers for self-assembly of plasmonic nanostructure.30-37 Because the oscillations of conduction electrons in metal NPs enhance the local fields in a small volume around them, these plasmonic nanostructures illustrate novel optical behavior such as circular dichroism, ultraviolet-visible spectrometry, surface enhanced Raman scattering (SERS) and so on.38-43 To study more novel optical behavior of the plasmonic nanostructures, the precise and highyield placement of NPs onto the DNA origami is of vital importance. AuNPs are among the most interested metal NPs due to their controlled size, various shape and high stability.44-46 To obtain AuNPs-based plasmonic nanostructures, the AuNPs are fully assembled with single stranded DNA and then be anchored to the DNA origami template at the predefined sites.47 In fact, ssDNA fully modified AuNPs, especially the large-sized AuNPs (≥ 30 nm) are not easy to be anchored onto the DNA origami templates because of the charge repulsion. Some previous works reported the employment of the hollow part on the triangular DNA origami or the edge sites of the DNA origami to anchor the large-sized AuNPs.48, 49 Herein, we used a cavity-type DNA origami as the template for the programmed positioning of 30 nm AuNPs. Different
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number of the handles around the cavity were examined by the hybridization of DNA-AuNPs. Thus prepared AuNPs dimer and tetramer nanostructure showed enhanced SERS performance.
EXPERIMENTAL SECTION Materials. All short oligo-DNA strands were purchased from Sangon (PAGE purification). Thiol-modified DNA was purchased from Takara (high-performance liquid chromatography purification). 30 nm gold nanoparticles were obtained by BBI. All of chemicals were supplied by Sigma. M13mp18 ss-DNA was purchased from New England Biolabs which was used as received. All short oligo-DNA strands were purchased from Sangon (PAGE purification). Thiolmodified DNA was purchased from Takara (high-performance liquid chromatography purification). 30 nm gold nanoparticles were obtained by BBI. All of chemicals were supplied by Sigma. M13mp18 ss-DNA was purchased from New England Biolabs which was used as received. Preparation of AuNPs-DNA Conjugates. 1 µL 100 nM thiol-modified DNA was added to 100 µL 30 nm AuNP solution in 0.5×TBE buffer (89 mM Tris, 89 mM boric acid, 2 mM EDTA, pH 8.0), incubated the mixture at 37 °C for four hours. 10 µL of 3M NaCl was added slowly into the reaction solution for 4 times in 2 hours with the final concentration of NaCl reached 300 mM. Then the solution was allowed to sit at 37 °C overnight. The DNA functionalized AuNPs were purified four times by centrifugations (7000 rpm, 10min). Each time, the supernatant was carefully removed and then the AuNPs were resuspended in a 0.5×TBE buffer to get rid of excess thiol-modified DNA. The concentration of AuNPs was measured using UV-Vis spectroscopy.
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Assembly of the DNA Origami Template. Cavity-type DNA origami template was obtained in a one-pot construction, in which 2 nM of M13mp18 DNA was mixed with 10 nM of staple strands in a 1 × TAE/Mg2+ (40 mM Tris-HCl, 20 mM Boric acid, 2 mM EDTA and 12.5 mM magnesium acetate, pH 8.0) buffer. They were annealed from 95 °C to 20 °C which was slowly decreasing the temperature at a rate of 1 °C min-1. To remove the extra strands, the prepared DNA origami was filtered by the 100 kDa (MWCO) centrifuge filters for four times. Assembly of AuNPs onto DNA Origami Template. The AuNPs were mixed with the cavitytype origami template at a molar ratio of 3:1 or 6:1. The assembly was performed by annealing the samples from 45 °C to 30 °C at a rate of 0.6 °C min-1 repeated four times and then cooling to 4 °C. AFM Characterization. The AFM (atomic force microscopy) images were obtained by a MultiMode 8 AFM with NanoScope V Controller (Bruker, Inc.). Samples were prepared by deposition of 2 µL onto freshly cleaved mica, and left to adsorb to the surface for 3min. After that, the sample was rinsed by water and dried by blowing nitrogen. They were scanned in tapping mode by AFM. TEM Characterization. For TEM imaging, 10 µL sample was dropped on a carbon-coated grid (400 mesh, Ted Pella). After 15 min deposition, the excess solution drop was wicked from the grid by absorption into filter paper. To remove the deposited salt, the grid was washed by a droplet of water and then used filter paper to wick the excess water away. The grid was kept at a bake oven for 4 hours for drying. The TEM characterization was conducted using a Tecnei G220S TWIN system, operated at 200 kV with a bright field mode. SERS Characterization. Samples (AuNPs, dimer-AuNPs and tetramer-AuNPs) were incubated in the 5 mM 4-Mercaptobenzoic acid (4-MBA, 99%) ethanolic solution for 2 h
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followed by a thorough rinse on pure ethanol. 10 mL of the mixture was deposited on a silicon substrate and the SERS characterization was conducted in liquid state. The SERS spectra were recorded by a confocal Raman microscope (In Via, Renishaw, England) equipped with a 633 nm He-Ne laser. A 20× objective, resolution grating of 1800 grooves, and a slit of 100 µM were used on all measurements. The spectra ranged from 1000 to 1850 cm−1. For all measurements, the experimental parameters are as follows: laser power 0.08 mW, acquisition time 10 s.
RESULTS AND DISCUSSION To obtain high yield of the AuNPs precisely anchored at the specific sites on the DNA origami template, the charge repulsion between DNA-AuNPs and DNA origami should be weakened at certain degree. Two typical factors should be carefully considered which are the density of ssDNA assembled on the surface of AuNPs and the design of the DNA origami. Compared to small-sized AuNPs, large-sized AuNPs own larger surface area which are much easier for the precipitation. In order to solve this problem, large-sized AuNPs should be assembled by much more ssDNA on their surface to keep stability, which would enlarge the charge repulsion between large-sized DNA-AuNPs and the DNA origami templates. Therefore, design of the DNA origami template to reduce such charge repulsion is of vital importance. In this study, we designed the cavities by dismissing several staples in the center part of the traditional rectangular DNA origami. The cavities were also rectangular in size of 12 nm * 15 nm, which supplied more space for the anchoring of 30 nm AuNPs. Beside the cavities, capture strands were set to the prolonged part of staples for the hybridization of the thiol-modified ssDNA on the surface of 30 nm AuNPs. Different number of ssDNA handles were studied for the anchoring of AuNPs which generated dimer- (Figure 1a) and tetramer-AuNPs (Figure 1b). As the dimer- and tetramer-
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AuNPs are both plasmonic nanostructures, their SERS properties are also investigated (Figure 1c). Schematic and AFM images in Figure 2 demonstrated the comparison of the cavity-type rectangular DNA origami and rectangular DNA origami (Figure S1). Typical TEM images in Figure 2 illustrated the successful assembly of dimer-AuNPs on a rectangular DNA origami platform with two cavities (Figure S2). To figure out the effect how the cavities can help for the anchoring of AuNPs, we fixed the size of the two cavities on the rectangular DNA origami while altered the number of capture strands. When two capture strands were set beside the cavities, the yield of dimer-AuNPs obtained on the cavity-type DNA origami templates was 37% which was a little higher than the yields obtained on the rectangle DNA origami templates (Figure 2a). There seemed no significant difference because two DNA capture strands for the anchoring of 30 nm AuNPs were unstable. As the capture strands were increased to 4, the yield of dimer-AuNPs was increased to 53% which was higher than 47% that obtained on the rectangle DNA origami template (Figure 2b). The promoted yields indicated that 30 nm AuNPs needed 4 capture strands at least for their organization onto the template. After the capture strands were increased to 6, the yield of dimer-AuNPs was up to 80%, which was much higher than 61% that obtained on the rectangle DNA origami template (Figure 2c). This data showed that the cavities helped to weaken the charge repulsion between DNA-AuNPs and DNA origami templates which could induce higher yields for the anchoring of 30 nm AuNPs. When the capture strands were increased to 8, the yield of dimer-AuNPs was 81% (Figure 2d). Compared to 76%, the difference between the yields obtained from the cavity-type and rectangle DNA origami templates became smaller. Also, the increased capture strands seemed not to help for the higher yields. Similar results were obtained by gel electrophoresis (Figure S3). This was because the increased capture
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strands seemed to full the cavities. The AFM images in Figure 2 illustrated the cavities in the center of rectangular DNA origami became more and more unapparent as the capture strands gradually increased. This may increase the charge repulsion again between DNA-AuNPs and DNA origami templates. In order to further verify that the cavity-type rectangular DNA origami templates were more efficient than rectangular DNA origami templates for the anchoring of the 30 nm AuNPs, we anchored four 30 nm AuNPs onto the cavity-type DNA origami template to form tetramerAuNPs. (Figure 3). We still used rectangular DNA origami as a control. Based on the experience for anchoring two 30 nm AuNPs on the cavity-type DNA origami, we selected six capture strands to localize individual AuNPs. Schematic and AFM images demonstrated the formation of rectangular DNA origami with and without four cavities (Figure S4). Typical TEM images in Figure 3 showed that 30 nm AuNPs were difficult to anchor onto the rectangular DNA origami template. The yield of thus prepared tetramer-AuNPs was 19%. This was because of the strong charge repulsion. But for the cavity-type rectangular DNA origami templates, AuNPs could be sufficiently assembled into tetramers (Figure S5) with the yield of 48%. Similar results were obtained by gel electrophoresis (Figure S6). These results showed that the cavity-type DNA origami was effective for the placement of large-sized AuNPs. The dimer- and tetramer-AuNPs impelled us to consider their potential plasmonic functionalities. Previous studies showed that metal assemblies with individual metal NPs in proper distances would generate stronger SERS signal. The distances between AuNPs in dimerand tetramer-AuNPs were designed to 20 nm in horizontal direction and 10 nm in vertical direction in this design which may generate the coupling of plasmons detected by UV–vis absorption (Figure S7). We employed 4-Mercaptobenzoic acid (4-MBA) served as a Raman-
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active molecule which could covalently attached the dimer- and tetramer-AuNPs by the strong interaction of Au-S bond. The frequency of 1580 cm-1 and 1075 cm-1 in the SERS spectra were clearly attributed to 4-MBA50, 51. Compared to individual AuNPs samples, the dimer- (red curve) and tetramer-AuNPs (blue curve) obtained significant enhanced signals under SERS detection (Figure 4). Taken the peak at 1075 cm−1 for example, the SERS signal intensity increased from 1633.6 to 5873.9 au with the increasing number of AuNPs in the nanostructures (Figure 5). This can be explained by the hot spots which generated by the small gaps between closely spaced NPs.
CONCLUSION In summary, we have developed a strategy by employing the cavity-type DNA origami for the effective anchoring of lager-size AuNPs. This strategy did not expense much time for design but dismiss several staples to form the cavities in a predefined DNA origami. The cavities could help to weaken the charge repulsion between DNA-AuNPs and DNA origami template which induced a precisely arrangement of lager-size AuNPs in high yield. Thus prepared dimer- and tetramerAuNPs were validated by TEM which demonstrated their organization and structural uniformity. Also, the dimer- and tetramer-AuNPs obtained significant enhanced signals under SERS detection. The methodology presented here opens up new possibilities to rationally engineer substrates based on DNA origami for optical and plasmonic applications.
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Figure 1. Schematic showing of AuNPs assemblies on the DNA origami form dimers (a) and tetramer structure (b). Typical SERS spectra of 4-Mercaptobenzoic acid (4-MBA) attached to metal nanoparticles assembled on DNA origami (c).
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Figure 2. Schematic illustration and AFM image of rectangular and cavity-type DNA origami which carried two (a), four (b), six (c) and eight (d) capture strands, respectively. Based on these templates, TEM images demonstrated the assembly of dimer-AuNPs with the statistical analysis of their yields. The scale bars are 200 nm.
Figure 3. Schematic illustration and TEM images of four 30 nm AuNPs’ anchoring based on the rectangular (a) and cavity-type (b) DNA origami templates with the statistical analysis of their yields (c). The scale bars are 100 nm.
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Figure 4. Typical SERS spectra of 4-MBA attached to individual AuNPs (black), dimer-AuNPs (red), tetramer-AuNPs (blue), respectively.
Figure 5. Changes of the characteristic SERS intensity at 1075 cm−1 of AuNPs, dimer-AuNPs and tetramer-AuNPs based on the cavity-type DNA origami with six capture strands.
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ASSOCIATED CONTENT Supporting Information. AFM images, TEM images, SERS intensity characterization and additional experimental data (PDF)
AUTHOR INFORMATION Corresponding Author *E-mail:
[email protected] *E-mail:
[email protected] Author Contributions The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript. ∥ M. Z. and X. W. contributed equally to this work. Notes The authors declare no competing financial interest.
ACKNOWLEDGMENT The authors acknowledge financial support from the National Natural Science Foundation of China (21475064), the Natural Science Foundation of Jiangsu Province (BK20151504), Program for Changjiang Scholars and Innovative Research Team in University (IRT_15R37), Sci-Tech Support Plan of Jiangsu Province (BE2014719), the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD, YX03001), the Mega-projects of Science
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and Technology
Research
(AWS13C007),
NUPTSF
(Grant
no.
214175),
MOST
(2016YFA0201200) and CAS (QYZDJ-SSW-SLH031).
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Table of Contents
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