Development of an advanced diagnostic concept for intestinal inflammation: molecular visualisation of nitric oxide in macrophages by functional poly(lactic-co-glycolic acid) microspheres

• Kathleen Lange,
• Christian Lautenschläger,
• Maria Wallert,
• Stefan Lorkowski,
• Andreas Stallmach and
• Alexander Schiller

Beilstein J. Nanotechnol. 2017, 8, 1637–1641, doi:10.3762/bjnano.8.163

• Salmonella typhimurium. After treatment with NO550 microparticles, only activated cells caused a green particle fluorescence and could be detected by laser scanning microscopy. NO release was confirmed indirectly with Griess reaction. Our functional NO550 particles enable a simple and early evaluation of

• -loaded microspheres. Confocal laser scanning microscopy (CLSM) images of activated (A) and inactivated (B) NO550-loaded polymeric microspheres in DPBS including fluorescence emission spectra (C) of the microspheres are shown. UV-irradiated SNP (10 mM, 2 min, 254 nm) was used for 2 minutes to activate

Uptake of the proteins HTRA1 and HTRA2 by cells mediated by calcium phosphate nanoparticles

• Olga Rotan,
• Katharina N. Severin,
• Simon Pöpsel,
• Alexander Peetsch,
• Melisa Merdanovic,
• Michael Ehrmann and
• Matthias Epple

Beilstein J. Nanotechnol. 2017, 8, 381–393, doi:10.3762/bjnano.8.40

• electron microscopy. Various cell types (HeLa, MG-63, THP-1, and hMSC) were incubated with fluorescently labelled proteins alone or with protein-loaded cationic and anionic nanoparticles. The cellular uptake was followed by light and fluorescence microscopy, confocal laser scanning microscopy (CLSM), and

• nm, emission: 460 nm) channels. Confocal laser scanning microscopy (CLSM) was performed on a Leica SP5 confocal inverse CLSM. The cells were stained with DAPI (nucleus) and Cell mask™ (cell membrane) to indicate the cellular uptake of the nanoparticles. To elucidate the uptake mechanism, several

On the pathway of cellular uptake: new insight into the interaction between the cell membrane and very small nanoparticles

• Claudia Messerschmidt,
• Daniel Hofmann,
• Anja Kroeger,
• Katharina Landfester,
• Volker Mailänder and
• Ingo Lieberwirth

Beilstein J. Nanotechnol. 2016, 7, 1296–1311, doi:10.3762/bjnano.7.121

• to have a detailed look at the very moment of particle uptake, revealing a remarkable dependency of the observed morphologies on particle size. Alongside the EM analysis we applied confocal laser scanning microscopy (CLSM), biochemical assays for LDH and ATP, flow cytometry, Caspase-3 western

• measurements. Preliminary, a qualitative examination of the cytotoxic effects was conducted by fluorescence activated cell sorting using FSC/SSC analysis (data not shown) and confocal laser scanning microscopy (Supporting Information File 1, Figure S10). Further experiments examined the LDH release and the

Functional diversity of resilin in Arthropoda

• Jan Michels,
• Esther Appel and
• Stanislav N. Gorb

Beilstein J. Nanotechnol. 2016, 7, 1241–1259, doi:10.3762/bjnano.7.115

• with a larger number of arthropod species, it will represent the first reliable method that specifically identifies resilin. The development and improvement of methods applying techniques such as micromechanical testing, atomic force microscopy and confocal laser scanning microscopy (CLSM) have

Reconstitution of the membrane protein OmpF into biomimetic block copolymer–phospholipid hybrid membranes

• Matthias Bieligmeyer,
• Franjo Artukovic,
• Stephan Nussberger,
• Thomas Hirth,
• Thomas Schiestel and
• Michaela Müller

Beilstein J. Nanotechnol. 2016, 7, 881–892, doi:10.3762/bjnano.7.80

• to retain the polymeric character of the membranes, the amount of lipid used was limited to 10 mol %. Eventually, membranes were analyzed using confocal laser scanning microscopy, voltage clamp measurements and impedance spectroscopy. Results and Discussion Block copolymer synthesis Poly(1,4-isoprene

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

• represented as the mean ± SD (n = 3). Confocal laser scanning microscopy images of human cells treated with calcitriol entrapped in C6-PLGA NPs. S2-013 cells: (A) control cells; cells after (B) 2 and (C) 72 h of incubation with C6–calcitriol–PLGA NPs. hTERT-HPNE cells: (D) control cells; cells after (E) 2 and

Mandibular gnathobases of marine planktonic copepods – feeding tools with complex micro- and nanoscale composite architectures

• Jan Michels and
• Stanislav N. Gorb

Beilstein J. Nanotechnol. 2015, 6, 674–685, doi:10.3762/bjnano.6.68

• with modern high-resolution elemental analysis techniques and confocal laser scanning microscopy [32][33][34] (Figures 3b, 4c–e, 5b,d), the results of high-resolution transmission electron microscopy analyses clearly indicate that the silica in the gnathobase teeth is composed of only some amorphous

Silica micro/nanospheres for theranostics: from bimodal MRI and fluorescent imaging probes to cancer therapy

• Shanka Walia and
• Amitabha Acharya

Beilstein J. Nanotechnol. 2015, 6, 546–558, doi:10.3762/bjnano.6.57

• liver cancer cells, NIH-3T3 mouse fibroblast cells, and 4T1 mouse breast cancer cells. Confocal laser scanning microscopy (CLSM) images indicated a successful bioconjugation of silica-coated QDs and MQDs with a bio-anchored membrane. Again, Salgueiriño-Maceira et al. [44] reported a new class 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

• (magnetite) embedded (PLLA–Fe). Confocal laser scanning microscopy (cLSM) was performed to determine the intracellular localization of the nanoparticles. The images in Figure 2 clearly show that all particle types were indeed internalized by the cells and not only attached to the cell surface. hHSCs showed a

• , apoptotic, dead) and taking the percentage of gated cells in the respective gate. Signals from APC-coupled antibodies were analyzed by overlays of several signals in die APC channel. Confocal laser scanning microscopy (cLSM) Confocal laser scanning microscopy was performed to validate the intracellular

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

• The extent of internalization of the different QDs and the Au@MnO particles was demonstrated by confocal laser scanning microscopy (Figure 4). After 24 h of incubation, all QD formulations were visible as red dots inside the cells (Figure 4a). Interestingly, the positively charged variant showed 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

• co-localized with marker proteins that help to identify both the uptaking cell type and its activation status (Figure 3b) [50][75][83]. Optical resolution of such co-localizations may be increased by various technical refinements, including confocal laser scanning microscopy which allows for serial

• additional advantages described for confocal laser scanning microscopy [119][120]. Of note, conventionally prepared slides can be used for SR-SIM and no special preparation is required [121]. We used this technique to localize FITC-labeled SiO2-NP in FFPE tissue sections of mice that were sliced and dewaxed

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

• ) showing an unchanged organic matrix structure after demineralization compared to the original mineralized nacre reference. Light microscopy and confocal laser scanning microscopy studies of stained samples show the presence of insoluble proteins at the chitin surface but not between the chitin layers

• light microscopy and confocal fluorescence laser scanning microscopy (FCLSM) as can be seen in Figure 3. A freshly broken cross section of original nacre was analyzed by SEM (see below in Figure 4c) and clearly reveals the layered structure of aragonite tablets. The insoluble organic matrix can be seen

• of the insoluble proteins with Coomassie blue, whereas the space in between the layers does not show any significant stain. The same observation can be made by fluorescence confocal laser scanning microscopy (Figure 3b) for which the thin cuts have been stained with rhodamine B ITC. Also in these

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

• nucleoside analogue to thymidine, into replicated DNA can be visualized by confocal laser scanning microscopy. As shown in Figure 3, cell proliferation was not significantly altered as shown by this assay. Responses of PCLS after exposure to (nano)particles Cytotoxic response After 4 h of incubation with 20

Interaction of dermatologically relevant nanoparticles with skin cells and skin

• Annika Vogt,
• Fiorenza Rancan,
• Sebastian Ahlberg,
• Berouz Nazemi,
• Chun Sik Choe,
• Maxim E. Darvin,
• Sabrina Hadam,
• Ulrike Blume-Peytavi,
• Kateryna Loza,
• Jörg Diendorf,
• Matthias Epple,
• Christina Graf,
• Eckart Rühl,
• Martina C. Meinke and
• Jürgen Lademann

Beilstein J. Nanotechnol. 2014, 5, 2363–2373, doi:10.3762/bjnano.5.245

• /streptomycin, 2% glutamine and 10% fetal calf serum. The cells grown in an incubator with 5% CO2, 100% humidity at 37 °C and incubated with the different silica particles (10 μg/mL) for 2 h. Analysis was performed by using flow cytometry and confocal laser scanning microscopy. Figure 3 was modified with

• ) or 75 nm (b,d) as well as negative (a,b) or positive (c,d) surface charge through funtionalization with (3-aminopropyl)triethoxysilane (APS) groups. Cells were incubated with particles (10 µg/mL, 2 h, 37 °C) and analyzed by means of flow cytometry (e,f) and confocal laser scanning microscopy (g

Anticancer efficacy of a supramolecular complex of a 2-diethylaminoethyl–dextran–MMA graft copolymer and paclitaxel used as an artificial enzyme

• Yasuhiko Onishi,
• Yuki Eshita,
• Rui-Cheng Ji,
• Masayasu Onishi,
• Takashi Kobayashi,
• Masaaki Mizuno,
• Jun Yoshida and
• Naoji Kubota

Beilstein J. Nanotechnol. 2014, 5, 2293–2307, doi:10.3762/bjnano.5.238

• represent an outstanding drug delivery method [1][2][3]. Recent detailed research of Maysinger et al. measuring the intracellular distribution of fluorescently labeled polymer micelles by using confocal laser scanning microscopy has shown the effect of a drug administered with a polymer DDS. As for polymer

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

• with specific organelle markers. Laser scanning microscopy and phase-contrast microscopy were performed in parallel on identical cell areas (Figure 1A). In cells cultured in the presence of Ag-NPs, agglomerated nanoparticles were visible in a region close to the cell nucleus but not in the cell culture

• uptake by laser scanning microscopy (LSM) LSM was performed to demonstrate the occurrence of intracellular silver nanoparticles in hMSCs after incubation. Therefore, hMSCs were subconfluently grown on 2-well Lab-TekTM glass chamber slides (Thermo Fisher Scientific, Langenselbold, Germany) and

• both ionic and nanoparticulate silver on the differentiation of human mesenchymal stem cells (hMSCs) into adipogenic, osteogenic and chondrogenic lineages and on the secretion of the respective differentiation markers adiponectin, osteocalcin and aggrecan. Results: As shown through laser scanning

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

• scanning microscopy) are other techniques that permit to visualize nanoparticles in cells. Therefore, hMSC were cultured in the presence of either 20 µg mL−1 silver (as nanoparticles) or 2 µg mL−1 silver ions (as silver acetate; control to separate the nanoparticle and the ion effect) at 37 °C for 24 h

• dissolution process in cells (including the localization of low concentrations of small nanoparticles as well as silver ions) imaging at the Ag L3,2 edges is a promising option for future work. Focused ion beam (FIB) and optical microscopy (phase contrast microscopy; fluorescence microscopy; confocal laser

Imaging the intracellular degradation of biodegradable polymer nanoparticles

• Anne-Kathrin Barthel,
• Martin Dass,
• Melanie Dröge,
• Jens-Michael Cramer,
• Daniela Baumann,
• Markus Urban,
• Katharina Landfester,
• Volker Mailänder and
• Ingo Lieberwirth

Beilstein J. Nanotechnol. 2014, 5, 1905–1917, doi:10.3762/bjnano.5.201

• ., number of detached magnetite crystals, and the number of nanoparticles in one endosome), we demonstrate the importance of TEM studies for such applications in addition to fluorescence studies (flow cytometry and confocal laser scanning microscopy). Keywords: biodegradation; mesenchymal stem cells; PLLA

• period of 14 days, primarily by means of transmission electron microscopy (TEM), in order to demonstrate their degradation. Furthermore, confocal laser scanning microscopy (CLSM) and flow cytometry were used to monitor the nanoparticle load of individual cells. As a probe we chose tailor-made PLLA

• cultures. All values are triplicates with the error bars representing the standard deviation. Confocal laser scanning microscopy (CLSM) Confocal laser scanning microscopy (CLSM) was applied to demonstrate the intracellular distribution of nanoparticles over the period of 14 days. As described in [26], for

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

• presence of the endocytotic proteins which are involved in endocytosis in both cell types (Figure 2). To achieve this, laser scanning microscopy (LSM) was applied as the primary tool for this investigations. Flotillin-1 and clathrin heavy chain could be visualized in J774A.1 cells, but caveolin-1 was not

• fluospheres (molecular probes) were used at a concentration of 20 µg/mL in RPMI. Laser scanning microscopy of fixed and living cells For LSM imaging, the cells were fixed with 3% paraformaldehyde (PFA, Sigma-Aldrich, Switzerland) in PBS for 15 minutes at room temperature. The cells were then washed with 1

• . Laser scanning microscopy imaging revealed particle uptake in J774A.1 and A549 cells. (A–C) Uptake of 40 nm PS NPs (NP: red, cytosol: grey). (A) Untreated cells with 40 nm NPs. (B) 40 nm NPs and cytochalasin D (cytoD) in J774A.1 and chlorpromazine (cpz) in A549 cells. (C) 40 nm NPs and

Mimicking exposures to acute and lifetime concentrations of inhaled silver nanoparticles by two different in vitro approaches

• Fabian Herzog,
• Kateryna Loza,
• Sandor Balog,
• Martin J. D. Clift,
• Matthias Epple,
• Peter Gehr,
• Alke Petri-Fink and
• Barbara Rothen-Rutishauser

Beilstein J. Nanotechnol. 2014, 5, 1357–1370, doi:10.3762/bjnano.5.149

• , resulting in increased surface concentrations of 1.7, 3.4, and 5.1 µg Ag/cm2. Therefore, the two exposure scenarios could be compared due to similar mass deposition on the lung cells surface. Cell morphology and particle uptake The cell morphology was studied with laser scanning microscopy (LSM) (Figure 2

• ; Sigma-Aldrich) served as positive control to induce the release of TNF-α and IL-8, respectively. Laser scanning microscopy As described in [44], the triple cell co-cultures were fixed on the cell culture insert with 3% paraformaldehyde in phosphate buffered saline (PBS) for 15 min at room temperature

Dry friction of microstructured polymer surfaces inspired by snake skin

• Martina J. Baum,
• Lars Heepe,
• Elena Fadeeva and
• Stanislav N. Gorb

Beilstein J. Nanotechnol. 2014, 5, 1091–1103, doi:10.3762/bjnano.5.122

• elevated, so the snake can generate propulsion due to the interlocking of its microstructure with surface asperities. The results of the study of the snake skin’s microstructure by using atomic force microscopy (AFM) and confocal laser scanning microscopy (CLSM) showed that the anisotropic geometry of the

Fibrillar adhesion with no clusterisation: Functional significance of material gradient along adhesive setae of insects

• Stanislav N. Gorb and
• Alexander E. Filippov

Beilstein J. Nanotechnol. 2014, 5, 837–845, doi:10.3762/bjnano.5.95

• revealed by confocal laser scanning microscopy (CLSM). This gradient is hypothesized to be an evolutionary optimization enhancing adaptation of adhesive pads to rough surfaces, while simultaneously preventing setal clusterisation. Such an optimisation presumably increases the performance of the adhesive