Frog tongue surface microstructures: functional and evolutionary patterns

• Thomas Kleinteich and
• Stanislav N. Gorb

Beilstein J. Nanotechnol. 2016, 7, 893–903, doi:10.3762/bjnano.7.81

• frogs with 4% Lugol’s iodine potassium iodide solution before we dissected the tongues. For this purpose, we followed the protocol by Metscher [33] but adjusted the staining duration to two weeks to allow the staining solution to diffuse deep into entire frog specimens. After staining, we dissected the

Atomic force microscopy as analytical tool to study physico-mechanical properties of intestinal cells

• Christa Schimpel,
• Oliver Werzer,
• Eleonore Fröhlich,
• Gerd Leitinger,
• Markus Absenger-Novak,
• Birgit Teubl,
• Andreas Zimmer and
• Eva Roblegg

Beilstein J. Nanotechnol. 2015, 6, 1457–1466, doi:10.3762/bjnano.6.151

• developed brush border (Figure 3A). In contrast, F-actin staining at the apex of M cells was markedly decreased due to a reduced or absent brush border (Figure 3B–D). Elasticity (force-indentation) measurements of Caco-2 cells and M cells Villin is not only involved in the formation and/or regulation of the

• hydrated and flexible cells, measurements were performed in a semi-dry state as demonstrated elsewhere [37][38]. Tetramethylrhodamine (TRITC)-phalloidin staining Visualization of the cytoskeletal F-actin network was performed using TRITC-phalloidin (Invitrogen GmbH, Darmstadt, Germany) in a similar manner

• the intense red F-actin staining. In contrast, M cells show a reduced/absent brush border indicated by a reduced F-actin labeling (B–D) (scale bar = 20 µm). Force–indentation curves and topographical images of a Caco-2 cell (A–C) and a M cell (D–F) classified into peripheral region/cell edge, nuclear

Peptide-equipped tobacco mosaic virus templates for selective and controllable biomineral deposition

• Klara Altintoprak,
• Axel Seidenstücker,
• Alexander Welle,
• Sabine Eiben,
• Petia Atanasova,
• Nina Stitz,
• Alfred Plettl,
• Joachim Bill,
• Hartmut Gliemann,
• Holger Jeske,
• Dirk Rothenstein,
• Fania Geiger and
• Christina Wege

Beilstein J. Nanotechnol. 2015, 6, 1399–1412, doi:10.3762/bjnano.6.145

• dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) (Figure 2a). The efficiency of peptide conjugation was determined by the ratio of the band intensities of modified and nonmodified CPs after Coomassie Blue staining. The binding efficiencies to individual CP subunits were ≈60% for all

• mg−1 cm−1 [94]) . For estimating concentrations of the different biotemplate rods, the band intensities of modified CPs and unmodified CPLys after SDS-PAGE separation and Comassie Blue staining were compared (see below). Electrophoretic analysis The modified CPs were analyzed by denaturing SDS-PAGE

PLGA nanoparticles as a platform for vitamin D-based cancer therapy

• Maria J. Ramalho,
• Joana A. Loureiro,
• Bárbara Gomes,
• Manuela F. Frasco,
• Manuel A. N. Coelho and
• M. Carmo Pereira

Beilstein J. Nanotechnol. 2015, 6, 1306–1318, doi:10.3762/bjnano.6.135

• endo-lysosomal compartments) for 1 h. The cells were washed with PBS and mounted on a glycerol-based medium with DAPI for nuclear staining. Acquisitions were performed with a Leica TCS SP5 II confocal laser scanning microscope (Leica Microsystems, Germany) in emission mode. Untreated cells were also

• harvested and fixed with 70% v/v ethanol. The cells were then stained with a DNA staining solution (0.1% v/v TritonX-100, 20 µg/mL PI and 35 µg/mL of RNase A in PBS) at a cell density of 106 cells/mL. FCM (FACSCalibur, BD Biosciences, CA, USA) was performed by plotting 12,000 gated events per sample. The

Pulmonary surfactant augments cytotoxicity of silica nanoparticles: Studies on an in vitro air–blood barrier model

• Jennifer Y. Kasper,
• Lisa Feiden,
• Maria I. Hermanns,
• Christoph Bantz,
• Michael Maskos,
• Ronald E. Unger and
• C. James Kirkpatrick

Beilstein J. Nanotechnol. 2015, 6, 517–528, doi:10.3762/bjnano.6.54

• quantification method for determination of the number of viable cells is cell staining with crystal violet (CV, purchased from Merck, 1407) [19]. Crystal violet (N-hexamethyl pararosaniline) is a monochromatic dye which stains cell nuclei. After fixation of NP-exposed cells they were incubated with 50 μL/96er

• –plain, which was apically applied to the A549, is depicted in red. ISO-HAS-1 underneath aSNP–plain stimulated A549, (Figure 7e–f) were counterstained (IF) for CD31 (Figure 7f). E-cadherin staining of A549 shows an inconsistent pattern. However, A549 as well as ISO-HAS-1 formed a confluent monolayer. No

• aSNPs displaying different surfaces (–plain, –NH2, –COOH; 5–100 µg/mL, untreated control (uc)). A: Cell viability (mitochondrial activity, MTS, % of untreated control (uc)); B: cell viability (crystal violet staining of nuclei, cell loss measurement, % of uc); C: LDH release (membrane integrity, % of

Hematopoietic and mesenchymal stem cells: polymeric nanoparticle uptake and lineage differentiation

• Ivonne Brüstle,
• Thomas Simmet,
• Gerd Ulrich Nienhaus,
• Katharina Landfester and
• Volker Mailänder

Beilstein J. Nanotechnol. 2015, 6, 383–395, doi:10.3762/bjnano.6.38

• -8 was not significantly altered in the presence of the polymeric nanoparticles (Supporting Information File 1, Figure S4). Differentiation behavior under nanoparticle influence Cytochemical staining of hMSCs For analyzing the particle influence on the differentiation capacity of hMSCs, the cells

• were incubated with 300 µg/mL nanoparticles for 24 h before starting the differentiation by providing the adequate differentiation medium. After 24 days, osteogenic and chondrogenic differentiation was analyzed by detecting alkaline phosphatase activity or methylene blue staining, respectively

• . Detection of adipogenic differentiation was performed after 4 weeks with Oil-Red O staining. In the presence of the nanoparticles, the differentiation into the three investigated lineages was not affected, as determined by cytochemical staining (Figure 5). The detection of the extracellular matrix for the

Comparative evaluation of the impact on endothelial cells induced by different nanoparticle structures and functionalization

• Lisa Landgraf,
• Ines Müller,
• Peter Ernst,
• Miriam Schäfer,
• Christina Rosman,
• Isabel Schick,
• Oskar Köhler,
• Hartmut Oehring,
• Vladimir V. Breus,
• Thomas Basché,
• Carsten Sönnichsen,
• Wolfgang Tremel and
• Ingrid Hilger

Beilstein J. Nanotechnol. 2015, 6, 300–312, doi:10.3762/bjnano.6.28

• -angle diamond knife (Diatome, Biel, Switzerland) in a Leica Ultracut S ultramicrotome was used to make ultrathin sections and then staining with freshly prepared uranyl acetate and lead citrate was performed. The sections were evaluated using a transmission electron microscope EM 902A (Zeiss, Germany

The effect of surface charge on nonspecific uptake and cytotoxicity of CdSe/ZnS core/shell quantum dots

• Vladimir V. Breus,
• Anna Pietuch,
• Marco Tarantola,
• Thomas Basché and
• Andreas Janshoff

Beilstein J. Nanotechnol. 2015, 6, 281–292, doi:10.3762/bjnano.6.26

• ) for 48 h. Immunostaining After washing with PBS, fixation was carried out by immersing the cells into a 20 °C cold acetone/methanol mixture (1:1 v/v) for 10 min. Afterwards, the cells were washed three times with PBS, the unspecific binding sites were blocked with FCS, and incubation in staining

• solutions was carried out according to the manufacturer’s recommendation: Alexa Fluor-conjugated IgG1 anti-tubulin (BD Bioscience, Heidelberg, Germany) from mouse was used for labeling microtubules, and 4’,6-diamidino-2-phenylindole (DAPI, Sigma-Aldrich, Seelze) for nucleus and DNA labeling. Staining was

• pellet was redissolved in 1 mL of distilled water. Fluorescence microscopy measurements For the fluorescent staining experiments, an upright Olympus fluorescence microscope Olympus BX51, with a 40× water-immersion objective (Olympus, Tokyo, Japan), equipped with a color camera (3 MP) was used. The

Overview about the localization of nanoparticles in tissue and cellular context by different imaging techniques

• Anja Ostrowski,
• Daniel Nordmeyer,
• Alexander Boreham,
• Cornelia Holzhausen,
• Lars Mundhenk,
• Christina Graf,
• Martina C. Meinke,
• Annika Vogt,
• Sabrina Hadam,
• Jürgen Lademann,
• Eckart Rühl,
• Ulrike Alexiev and
• Achim D. Gruber

Beilstein J. Nanotechnol. 2015, 6, 263–280, doi:10.3762/bjnano.6.25

• -inflammatory effects with rather specific accumulation in inflamed tissues [55][56][57]. Due to their sulfate groups and the specific staining properties of Alcian blue, this method has been used successfully, for example, for the detection of dPGS amine accumulated in Kupffer cells in the liver of mice

• using full-field high-resolution transmission X-ray microscopy combined with a potassium permanganate staining of FFPE-tissue sections of the cerebellum and the liver [145]. Soft X-ray microscopy has been successfully applied for 3D imaging of vitrified cells without any further staining [146]. That

• example, titanium dioxide, SiO2-NP or QD, TEM has been widely used to characterize the morphology and size of NP as well as their location in tissues [28][35][39][113][156][157][158]. It has to be kept in mind, however, that artifacts due to staining with lead citrate and uranyl acetate can easily be

Oxygen-plasma-modified biomimetic nanofibrous scaffolds for enhanced compatibility of cardiovascular implants

• Anna Maria Pappa,
• Varvara Karagkiozaki,
• Silke Krol,
• Spyros Kassavetis,
• Dimitris Konstantinou,
• Charalampos Pitsalidis,
• Lazaros Tzounis,
• Nikos Pliatsikas and
• Stergios Logothetidis

Beilstein J. Nanotechnol. 2015, 6, 254–262, doi:10.3762/bjnano.6.24

• to the reference wavelength of 690 nm. Data (n = 3) were presented as means of O.D. values as well as normalized according to the control and presented as % cell viability. Optical imaging through methylene blue staining: Once fibroblasts were seeded onto either unmodified or surface-modified

Mechanical properties of MDCK II cells exposed to gold nanorods

• Anna Pietuch,
• Bastian Rouven Brückner,
• David Schneider,
• Marco Tarantola,
• Christina Rosman,
• Carsten Sönnichsen and
• Andreas Janshoff

Beilstein J. Nanotechnol. 2015, 6, 223–231, doi:10.3762/bjnano.6.21

• number of CTAB molecules which can, in principle, be released from the particle surface assuming a concentration of 12 μg/mL, we only found a Young’s modulus of E = 6.5 ± 1 kPa. Fluorescence microscopy images (staining of actin and microtubules) as well as AFM images (topography) reveal that the cells

• with PBS. Incubation with staining solution was carried out following the manufacture’s recommendation. For F-actin staining Alexa Fluor 546 Phalloidin (Invitrogen, Darmstadt, Germany) and for microtubules labeling Alexa 488 conjugated mouse anti-β-tubulin (BD Biosciences, Heidelberg, Germany) was used

• . Nucleus staining was carried out with DAPI (4’,6-diamidino-2-phenylindol, 50 ng/mL in PBS) (Sigma-Aldrich, Steinheim, Germany). Dark-field microscopy was carried out with an upright microscope with dark-field condensor (Olympus BX51) equipped with a 40× water immersion objective. AFM imaging and force

Multifunctional layered magnetic composites

• Maria Siglreitmeier,
• Baohu Wu,
• Tina Kollmann,
• Martin Neubauer,
• Gergely Nagy,
• Dietmar Schwahn,
• Vitaliy Pipich,
• Damien Faivre,
• Dirk Zahn,
• Andreas Fery and
• Helmut Cölfen

Beilstein J. Nanotechnol. 2015, 6, 134–148, doi:10.3762/bjnano.6.13

• to the matrix, which might derive from the usage of staining media or dehydration. For comparison studies, the structure of the original nacre matrix (Haliotis laevigata) was analyzed as well. Figure 2 represents very-small (VSANS) and small (SANS) angle neutron scattering profiles of nacre (top) and

• studies no staining of proteins in between the layers could be observed. Therefore we conclude that the insoluble matrix proteins are dominantly located directly at the β-chitin matrix and are not present in between the layers. Figure 3c and Figure 3d show an embedded sample of insoluble nacre matrix

• infiltrated with gelatin by a vacuum infiltration process. Staining of this sample illustrates not only blue stained chitin layers and insoluble matrix proteins but also colored areas in between the layers, indicating a filling of the matrix with gelatin. The interaction and positive stain of gelatin and

Mammalian cell growth on gold nanoparticle-decorated substrates is influenced by the nanoparticle coating

• Christina Rosman,
• Sebastien Pierrat,
• Marco Tarantola,
• David Schneider,
• Eva Sunnick,
• Andreas Janshoff and
• Carsten Sönnichsen

Beilstein J. Nanotechnol. 2014, 5, 2479–2488, doi:10.3762/bjnano.5.257

• staining test for live/dead should be performed. This was not performed since our results already indicated that CTAB nanorods are not suitable for live cell applications due to their impact on the native cell behavior. However, although the proliferation behavior of the tracked cells was poor, active cell

Proinflammatory and cytotoxic response to nanoparticles in precision-cut lung slices

• Stephanie Hirn,
• Nadine Haberl,
• Kateryna Loza,
• Matthias Epple,
• Wolfgang G. Kreyling,
• Barbara Rothen-Rutishauser,
• Markus Rehberg and
• Fritz Krombach

Beilstein J. Nanotechnol. 2014, 5, 2440–2449, doi:10.3762/bjnano.5.253

• , and dendritic cells, in their natural context has been shown in several studies by immunohistochemical staining [7][8][9]. Nevertheless, this in vitro system has also its limitations. There are obvious disadvantages in comparison to the in vivo situation such as no ventilation, no stretching, and no

• investigate whether PCLS have the potential to serve as a generally applicable effective tool in nanotoxicology. First, we determined the viability of PCLS up to 72 h by carrying out live/dead staining, lactate dehydrogenase (LDH) assay, and WST-1 assay. Second, we assessed the cytotoxic and proinflammatory

• staining, and WST-1 assay. As LDH is present in the cytoplasm of cells, detection of LDH in the culture medium of PCLS indicates a loss of cell membrane integrity. Therefore, LDH release is a direct measure of a cytotoxic response or an indirect measure of cell viability. As displayed in Figure 1, the LDH

Nanoparticle interactions with live cells: Quantitative fluorescence microscopy of nanoparticle size effects

• Li Shang,
• Karin Nienhaus,
• Xiue Jiang,
• Linxiao Yang,
• Katharina Landfester,
• Volker Mailänder,
• Thomas Simmet and
• G. Ulrich Nienhaus

Beilstein J. Nanotechnol. 2014, 5, 2388–2397, doi:10.3762/bjnano.5.248

• NPs in each cell was obtained by normalizing the integrated intensity by the cell area. The cell membrane and the intracellular region were identified based on the membrane staining. Schematic representation of the cellular uptake of (a) large and (b) small NPs. Whereas larger NPs exert interactions

Coating with luminal gut-constituents alters adherence of nanoparticles to intestinal epithelial cells

• Heike Sinnecker,
• Katrin Ramaker and
• Andreas Frey

Beilstein J. Nanotechnol. 2014, 5, 2308–2315, doi:10.3762/bjnano.5.239

• grown on transwell filters (Figure 3A, B; counter-staining of the cell surface with a lectin). Pretreatment with the protein BSA appears to alleviate agglomeration as well as adherence. Only few distinct spots are visible on the cell surface, compared to a pronounced adherence of markedly larger

Effect of silver nanoparticles on human mesenchymal stem cell differentiation

• Christina Sengstock,
• Jörg Diendorf,
• Matthias Epple,
• Thomas A. Schildhauer and
• Manfred Köller

Beilstein J. Nanotechnol. 2014, 5, 2058–2069, doi:10.3762/bjnano.5.214

• used (Figure 2B). To confirm adipogenic differentiation, the lipid content of the cells was visualized by using oil red O or Bodipy493/503 staining. As shown in Figure 3C, hMSCs differentiated into adipocytes in the presence of adipogenic-differentiation media (positive control; Figure 3C), in contrast

• ) revealed a decrease in lipid vacuoles with increasing silver concentrations. This decrease was significant at the applied concentrations of 10 µg·mL−1 for Ag-NP (black bars) or 1.0 µg·mL−1 for Ag+ ions (grey bars). The differences between oil red extraction and phase analysis after Bodipy493/503 staining

• of Ag-NP/Ag+ ions on the osteogenic differentiation of hMSCs was investigated after a period of 21 d. To confirm the differentiation of the Ag-NP/Ag+ ion-treated hMSCs into osteoblasts, alizarin red S staining was carried out to verify the mineralization of the cells. hMSCs exposed to 10 µg·mL−1 Ag

PVP-coated, negatively charged silver nanoparticles: A multi-center study of their physicochemical characteristics, cell culture and in vivo experiments

• Sebastian Ahlberg,
• Alexandra Antonopulos,
• Jörg Diendorf,
• Ralf Dringen,
• Matthias Epple,
• Rebekka Flöck,
• Wolfgang Goedecke,
• Christina Graf,
• Nadine Haberl,
• Jens Helmlinger,
• Fabian Herzog,
• Frederike Heuer,
• Stephanie Hirn,
• Christian Johannes,
• Stefanie Kittler,
• Manfred Köller,
• Katrin Korn,
• Wolfgang G. Kreyling,
• Fritz Krombach,
• Jürgen Lademann,
• Kateryna Loza,
• Eva M. Luther,
• Marcelina Malissek,
• Martina C. Meinke,
• Daniel Nordmeyer,
• Anne Pailliart,
• Jörg Raabe,
• Fiorenza Rancan,
• Barbara Rothen-Rutishauser,
• Eckart Rühl,
• Carsten Schleh,
• Andreas Seibel,
• Christina Sengstock,
• Lennart Treuel,
• Annika Vogt,
• Katrin Weber and
• Reinhard Zellner

Beilstein J. Nanotechnol. 2014, 5, 1944–1965, doi:10.3762/bjnano.5.205

• demonstrated for hMSC, primary T-cells, primary monocytes, and astrocytes. A visualization of particles inside cells is possible by X-ray microscopy, fluorescence microscopy, and combined FIB/SEM analysis. By staining organelles, their localization inside the cell can be additionally determined. While primary

• (TEM), this technique requires thin samples and, hence, slicing of the samples as well as additional staining, which both might change the properties of the samples. Furthermore, energy dispersive X-ray spectroscopy (EDX) combined with TEM has only limited spectral and spatial resolution and

• used as a tool to detect DNA-DSB by specific antibodies recognizing γ-H2AX. Staining of cells with antibodies directed against γ-H2AX results in a speckled staining of the nucleus. It is generally accepted that a single focus is representing a single DSB [118]. In the following, we describe experiments

Biocompatibility of cerium dioxide and silicon dioxide nanoparticles with endothelial cells

• Claudia Strobel,
• Martin Förster and
• Ingrid Hilger

Beilstein J. Nanotechnol. 2014, 5, 1795–1807, doi:10.3762/bjnano.5.190

• markers, respectively. After staining both CD31 (monoclonal anti-human CD31 antibodies conjugated to fluorescein isothiocyanate (FITC), Miltenyi Biotec GmbH, Germany) and CD90 (monoclonal anti-human CD90 antibodies conjugated to R-phycoerythrin (PE) Miltenyi Biotec GmbH, Germany), the cells were washed

• . As a permeabilization reagent, 0.1% (v/v) Saponin (Sigma-Aldrich Chemie GmbH, Germany) in Hank’s BSS was used. Intracellular staining of vWF with allophycocyanin (APC) conjugated mouse monoclonal anti-human vWF-A2 antibodies (R&D Systems, Inc., USA) followed. Unstained cells, cells stained with mouse

The surface properties of nanoparticles determine the agglomeration state and the size of the particles under physiological conditions

• Christoph Bantz,
• Olga Koshkina,
• Thomas Lang,
• Hans-Joachim Galla,
• C. James Kirkpatrick,
• Roland H. Stauber and
• Michael Maskos

Beilstein J. Nanotechnol. 2014, 5, 1774–1786, doi:10.3762/bjnano.5.188

• staining procedures might cause additional artifacts and furthermore also lead to an increase in preparation time, no staining was applied here and for the POS particles the TEM investigation is only useful at the first stage of the modification procedure. To compensate for the mentioned drawbacks of the

Influence of surface-modified maghemite nanoparticles on in vitro survival of human stem cells

• Michal Babič,
• Daniel Horák,
• Lyubov L. Lukash,
• Tetiana A. Ruban,
• Yurii N. Kolomiets,
• Svitlana P. Shpylova and
• Oksana A. Grypych

Beilstein J. Nanotechnol. 2014, 5, 1732–1737, doi:10.3762/bjnano.5.183

• medium. Confocal micrographs of 4BL human stem cells treated with (a, b) D-mannose-coated γ-Fe2O3, (c, d) PDMAAm-coated γ-Fe2O3 and (e, f) non-coated γ-Fe2O3 nanoparticles. Staining with DAPI and ThR. Scale bars: 10 μm. Acknowledgements Financial support of Ministry of Education, Youth and Sports of the

In vitro and in vivo interactions of selected nanoparticles with rodent serum proteins and their consequences in biokinetics

• Wolfgang G. Kreyling,
• Stefanie Fertsch-Gapp,
• Martin Schäffler,
• Blair D. Johnston,
• Nadine Haberl,
• Christian Pfeiffer,
• Jörg Diendorf,
• Carsten Schleh,
• Stephanie Hirn,
• Manuela Semmler-Behnke,
• Matthias Epple and
• Wolfgang J. Parak

Beilstein J. Nanotechnol. 2014, 5, 1699–1711, doi:10.3762/bjnano.5.180

• protein densitometry. Only predominantly binding serum proteins could be analyzed by this quantitative approach. A typical electrophoresis gel visualized by Coomassie staining is given in Figure 2, which shows a variety of proteins present in the corona of the three differently sized AuNP. Proteins cover

Different endocytotic uptake mechanisms for nanoparticles in epithelial cells and macrophages

• Dagmar A. Kuhn,
• Dimitri Vanhecke,
• Benjamin Michen,
• Fabian Blank,
• Peter Gehr,
• Alke Petri-Fink and
• Barbara Rothen-Rutishauser

Beilstein J. Nanotechnol. 2014, 5, 1625–1636, doi:10.3762/bjnano.5.174

• dehydrogenase (LDH) assay (see Figure S1, Supporting Information File 1). Trypan blue staining marked the integrity of the cell membrane for cells which were impaired by the inhibitor (red insets, Figure 3) and revealed the following percentage of dead cells for J774A.1 cells (n = 3): negative control 20% (SD

• affected the cells are summarized in Table 1 and the images in Figure 3 are presented with either red (impairment by inhibitor) or green (no effect by inhibitor) letter insets. Trypan blue staining demonstrated the same outcome. The percentage of dead cells which were treated with cytochalasin D was not as

• , flotillin-1 and caveolin-1 (all fluorescently labelled with Alexa Fluor 488, antibodies-online GmbH, Aachen, Germany) were used at a final dilution of 1:20 in 1× PBS on fixed cells. After 1 hour of staining (in a dark room at room temperature) and three washing cycles with 1× PBS, the cells were mounted

Protein-coated pH-responsive gold nanoparticles: Microwave-assisted synthesis and surface charge-dependent anticancer activity

• Dickson Joseph,
• Nisha Tyagi,
• Christian Geckeler and
• Kurt E.Geckeler

Beilstein J. Nanotechnol. 2014, 5, 1452–1462, doi:10.3762/bjnano.5.158

• ). The BSA-AuNP sample was mixed with 2× sample buffer (Sigma) and heated at 80 °C for 15 min. Further protein was separated through a 4% stacking gel and 12% separation gel. To determine the amount of protein in the sample, the gels were stained with Coomassie blue (Sigma) staining solution for 2 h. The

Near-field photochemical and radiation-induced chemical fabrication of nanopatterns of a self-assembled silane monolayer

• Ulrich C. Fischer,
• Carsten Hentschel,
• Florian Fontein,
• Linda Stegemann,
• Christiane Hoeppener,
• Harald Fuchs and
• Stefanie Hoeppener

Beilstein J. Nanotechnol. 2014, 5, 1441–1449, doi:10.3762/bjnano.5.156

• , respectively. The 1.2 µm structure is clearly resolved in the fluorescence image, but due to the diffraction limit the 0.22 µm pattern cannot be resolved. Only the defect structures of the periodic nanopatterns are recognized. In addition to the staining of the chemical nanostructures with fluorescein

• (a) 1.2 µm latex beads and of (b) 220 nm latex beads after staining of the chemically functionalized areas with fluorescein molecules. Optically only the larger hexagonal nanopattern can be resolved, whereas the fluorescence image of the 220 nm pattern is characterized by defect structures. (c) AFM