Substrate microtopographies induce cellular alignment and affect nuclear force transduction.

Cellular physiology has been mainly studied by using two-dimensional cell culture substrates which lack in vivo-mimicking extracellular environment and interactions. Thus, there is a growing need for more complex model systems in life sciences. Micro-engineered scaffolds have been proven to be a promising tool in understanding the role of physical cues in the co-regulation of cellular functions. These tools allow, for example, probing cell morphology and migration in response to changes in chemo-physical properties of their microenvironment. In order to understand how microtopographical features, what cells encounter in vivo, affect cytoskeletal organization and nuclear mechanics, we used direct laser writing via two-photon polymerization (TPP) to fabricate substrates which contain different surface microtopographies. By combining with advanced high-resolution spectral imaging, we describe how the constructed grid and vertical line microtopographies influence cellular alignment, nuclear morphology and mechanics. Specifically, we found that growing cells on grids larger than 10 × 20 µm2 and on vertical lines increased 3D actin cytoskeleton orientation along the walls of microtopographies and abolished basal actin stress fibers. In concert, the nuclei of these cells were also more aligned, elongated, deformed and less flattened, indicating changes in nuclear force transduction. Importantly, by using fluorescence lifetime imaging microscopy for measuring Förster resonance energy transfer for a genetically encoded nesprin-2 molecular tension sensor, we show that growing cells on these microtopographic substrates induce lower mechanical tension at the nuclear envelope. To conclude, here used substrate microtopographies modulated the cellular mechanics, and affected actin organization and nuclear force transduction.

Surface Passivation of III-V GaAs Nanopillars by Low-Frequency Plasma Deposition of Silicon Nitride for Active Nanophotonic Devices.

Numerous efforts have been devoted to improve the electronic and optical properties of III-V compound materials via reduction of their nonradiative states, aiming at highly efficient III-V sub-micrometer active devices and circuits. Despite many advances, the poor reproducibility and short-term passivation effect of chemical treatments, such as sulfidation and nitridation, requires the use of protective encapsulation methods, not only to protect the surface, but also to provide electrical isolation for device manufacturing. There is still a controversial debate on which combination of chemical treatment and capping dielectric layer can best reproducibly protect the crystal surface of III-V materials while being compatible with readily available semiconductor-foundry plasma deposition methods. This work reports on a systematic experimental study on the role of sulfide ammonium chemical treatment followed by dielectric coating (either silicon oxide or nitride) in the passivation effect of GaAs/AlGaAs nanopillars. Our results conclusively show that, under ambient conditions, the best surface passivation is achieved using ammonium sulfide followed by encapsulation with a thin layer of silicon nitride by low-frequency plasma-enhanced chemical deposition. Here, the sulfurized GaAs surfaces, high level of hydrogen ions, and low-frequency (380 kHz) excitation plasma that enable intense bombardment of hydrogen, all seem to provide a combined active role in the passivation mechanism of the pillars by reducing the surface states. As a result, we observe up to a 29-fold increase of the photoluminescence (PL) integrated intensity for the best samples as compared to untreated nanopillars. X-ray photoelectron spectroscopy analysis confirms the best treatments show remarkable removal of gallium and arsenic native oxides. Time-resolved micro-PL measurements display nanosecond lifetimes resulting in a record-low surface recombination velocity of ∼1.1 × 104 cm s-1 for dry-etched GaAs nanopillars. We achieve robust, stable, and long-term passivated nanopillar surfaces, which creates expectations for remarkable high internal quantum efficiency (IQE > 0.5) in nanoscale light-emitting diodes. The enhanced performance paves the way to many other nanostructures and devices such as miniature resonators, lasers, photodetectors, and solar cells, opening remarkable prospects for GaAs active nanophotonic devices.

3D Polymer Architectures for the Identification of Optimal Dimensions for Cellular Growth of 3D Cellular Models.

Organ-on-chips and scaffolds for tissue engineering are vital assay tools for pre-clinical testing and prediction of human response to drugs and toxins, while providing an ethical sound replacement for animal testing. A success criterion for these models is the ability to have structural parameters for optimized performance. Here we show that two-photon polymerization fabrication can create 3D test platforms, where scaffold parameters can be directly analyzed by their effects on cell growth and movement. We design and fabricate a 3D grid structure, consisting of wall structures with niches of various dimensions for probing cell attachment and movement, while providing easy access for fluorescence imaging. The 3D structures are fabricated from bio-compatible polymer SZ2080 and subsequently seeded with A549 lung epithelia cells. The seeded structures are imaged with confocal microscopy, where spectral imaging with linear unmixing is used to separate auto-fluorescence scaffold contribution from the cell fluorescence. The volume of cellular material present in different sections of the structures is analyzed, to study the influence of structural parameters on cell distribution. Furthermore, time-lapse studies are performed to map the relation between scaffold parameters and cell movement. In the future, this kind of differentiated 3D growth platform, could be applied for optimized culture growth, cell differentiation, and advanced cell therapies.

Design and manufacture of an all-polymeric integrated multimode interferometer for visible photonics.

This work demonstrates an integrated multimode interferometer (MMI) based on a fully polymeric platform and optimized for visible range operation. The dimensions of a 2×2 MMI are first calculated analytically and simulated using finite elements method. The devices are manufactured using two layers of negative tone photoresists. The top layer is patterned by e-beam lithography demonstrating the adaptability of this material, naturally designed to respond to UV radiation. Fabrication tolerance was smaller than 100 nm. Devices were optically characterized with a 635 nm input source and the best performance for a 3 dB power splitter was found at an interferometric cavity dimension of 10.5 × 190.68 µm. Other interferometric lengths were characterized to establish a process design kit that allows future use of this platform in more complex photonic integrated circuits architectures.

Spectral imaging and nucleic acid mimics fluorescence in situ hybridization (SI-NAM-FISH) for multiplex detection of clinical pathogens.

The application of nucleic acid mimics (NAMs), such as locked nucleic acid (LNA) and 2'-O-methyl-RNA (2'OMe), has improved the performance of fluorescence in situ hybridization (FISH) methods for the detection/location of clinical pathogens since they provide design versatility and thermodynamic control. However, an important limitation of FISH techniques is the low number of distinguishable targets. The use of filters in fluorescence image acquisition limits the number of fluorochromes that can be simultaneously differentiated. Recent advances in fluorescence spectral image acquisition have allowed the unambiguous identification of several microorganisms in a single sample. In this work, we aimed to combine NAM-FISH and spectral image analysis to develop and validate a new FISH variant, the spectral imaging-NAM-FISH (SI-NAM-FISH), that allows a multiplexed, robust and rapid detection of clinical pathogens. In the first stage, to implement/validate the method, we have selected seven fluorochromes with distinct spectral properties and seven bacterial species (Pseudomonas aeruginosa, Citrobacter freundii, Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, and Acinetobacter calcoaceticus). As a strong variation in fluorescence intensities is found between species and between fluorochromes, seven versions of a EUB LNA/2'OMe probe, each conjugated to one of seven fluorochromes, were used to rank species/fluorochromes by FISH and then optimize species/fluorochrome pairing. Then, final validation tests were performed using mixed populations to evaluate the potential of the technique for separating/quantifying the different targets. Overall, validation tests with different proportions of bacteria labeled with the respective fluorochrome have shown the ability of the method to correctly distinguish the species.

Spectral-temporal luminescence properties of Colloidal CdSe/ZnS Quantum Dots in relevant polymer matrices for integration in low turn-on voltage AC-driven LEDs.

This work employs spectral and spectral-temporal Photoluminescence (PL) spectroscopy techniques to study the radiative mechanisms in colloidal CdSe/ZnS Quantum Dot (QD) thin films without and with 1% PMMA polymer matrix embedding (QDPMMA). The observed bimodal transient-spectral PL distributions reveal bandgap transitions and radiative recombinations after interdot electron transfer. The PMMA polymer embedding protects the QDs during the plasma-sputtering of inorganic layers electroluminescent (EL) devices, with minimal impact on the charge transfer properties. Further, a novel TiO2-based, all-electron bandgap, AC-driven QLED architecture is fabricated, yielding a surprisingly low turn-on voltage, with PL-identical and narrow-band EL emission. The symmetric TiO2 bilayer architecture is a promising test platform for alternative optical active materials.

Cellular Interaction of Bone Marrow Mesenchymal Stem Cells with Polymer and Hydrogel 3D Microscaffold Templates.

Biomimicking biological niches of healthy tissues or tumors can be achieved by means of artificial microenvironments, where structural and mechanical properties are crucial parameters to promote tissue formation and recreate natural conditions. In this work, three-dimensional (3D) scaffolds based on woodpile structures were fabricated by two-photon polymerization (2PP) of different photosensitive polymers (IP-S and SZ2080) and hydrogels (PEGDA 700) using two different 2PP setups, a commercial one and a customized one. The structures' properties were tuned to study the effect of scaffold dimensions (gap size) and their mechanical properties on the adhesion and proliferation of bone marrow mesenchymal stem cells (BM-MSCs), which can serve as a model for leukemic diseases, among other hematological applications. The woodpile structures feature gap sizes of 25, 50, and 100 µm and a fixed beam diameter of 25 µm, to systematically study the optimal cell colonization that promotes healthy cell growth and potential tissue formation. The characterization of the scaffolds involved scanning electron microscopy and mechanical nanoindenting, while their suitability for supporting cell growth was evaluated with live/dead cell assays and multistaining 3D confocal imaging. In the mechanical assays of the hydrogel material, we observed two different stiffness ranges depending on the indentation depth. Larger gap woodpile structures coated with fibronectin were identified as the most promising scaffolds for 3D BM-MSC cellular models, showing higher proliferation rates. The results indicate that both the design and the employed materials are suitable for further assays, where retaining the BM-MSC stemness and original features is crucial, including studies focused on BM disorders such as leukemia and others. Moreover, the combination of 3D scaffold geometry and materials holds great potential for the investigation of cellular behaviors in a co-culture setting, for example, mesenchymal and hematopoietic stem cells, to be further applied in medical research and pharmacological studies.

Two-photon polymerization simulation and fabrication of 3D microprinted suspended waveguides for on-chip optical interconnects.

Quantum and neuromorphic computational platforms in integrated photonic circuits require next-generation optical functionalities. Often, increasingly complex on-chip light-routing that allow superpositions not attainable by planar technologies are paramount e.g. for artificial neural networks. Versatile 3D waveguides are achievable via two-photon polymerization (TPP)-based microprinting. Here, a 3D morphology prediction tool which considers experimental TPP parameters, is presented, enabling on-chip 3D waveguide performance simulations. The simulations allow reducing the cost-intensive systematic experimental optimization process. Fabricated 3D waveguides show optical transmission properties in agreement with simulations, demonstrating that the developed morphology prediction methodology is beneficial for the development of versatile on-chip and potentially inter-chip photonic interconnect technology.

Photonic polymeric structures and electrodynamics simulation method based on a coupled oscillator finite-difference time-domain (O-FDTD) approach.

We use femtosecond laser-based two-photon polymerization (TPP) to fabricate a 2.5D micropillar array. Using an angular detection setup, we characterize the structure's scattering properties and compare the results against simulation results obtained from a novel electrodynamics simulation method. The algorithm employs a modified formulation of the Lorentz Oscillator Model and a leapfrog time differentiation to define a 2D coupled Oscillator Finite-Difference Time-Domain (O-FDTD). We validate the model by presenting several simulation examples that cover a wide range of photonic components, such as multi-mode interference splitters, photonic crystals, ring resonators, and Mach-Zehnder interferometers.

Cityscape LoRa Signal Propagation Predicted and Tested Using Real-World Building-Data Based O-FDTD Simulations and Experimental Characterization.

The age of the Internet of Things (IoT) and smart cities calls for low-power wireless communication networks, for which the Long-Range (LoRa) is a rising star. Efficient network engineering requires the accurate prediction of the Received Signal Strength Indicator (RSSI) spatial distribution. However, the most commonly used models either lack the physical accurateness, resolution, or versatility for cityscape real-world building distribution-based RSSI predictions. For this purpose, we apply the 2D electric field wave-propagation Oscillator Finite-Difference Time-Domain (O-FDTD) method, using the complex dielectric permittivity to model reflection and absorption effects by concrete walls and the receiver sensitivity as the threshold to obtain a simulated coverage area in a 600 × 600 m2 square. Further, we report a simple and low-cost method to experimentally determine the signal coverage area based on mapping communication response-time delays. The simulations show a strong building influence on the RSSI, compared against the Free-Space Path (FSPL) model. We obtain a spatial overlap of 84% between the O-FDTD simulated and experimental signal coverage maps. Our proof-of-concept approach is thoroughly discussed compared to previous works, outlining error sources and possible future improvements. O-FDTD is demonstrated to be most promising for both indoors and outdoors applications and presents a powerful tool for IoT and smart city planners.

Lipid Nanosystems and Serum Protein as Biomimetic Interfaces: Predicting the Biodistribution of a Caffeic Acid-Based Antioxidant.

PURPOSE: AntiOxCIN3 is a novel mitochondriotropic antioxidant developed to minimize the effects of oxidative stress on neurodegenerative diseases. Prior to an investment in pre-clinical in vivo studies, it is important to apply in silico and biophysical cell-free in vitro studies to predict AntiOxCIN3 biodistribution profile, respecting the need to preserve animal health in accordance with the EU principles (Directive 2010/63/EU). Accordingly, we propose an innovative toolbox of biophysical studies and mimetic models of biological interfaces, such as nanosystems with different compositions mimicking distinct membrane barriers and human serum albumin (HSA). METHODS: Intestinal and cell membrane permeation of AntiOxCIN3 was predicted using derivative spectrophotometry. AntiOxCIN3 -HSA binding was evaluated by intrinsic fluorescence quenching, synchronous fluorescence, and dynamic/electrophoretic light scattering. Steady-state and time-resolved fluorescence quenching was used to predict AntiOxCIN3-membrane orientation. Fluorescence anisotropy, synchrotron small- and wide-angle X-ray scattering were used to predict lipid membrane biophysical impairment caused by AntiOxCIN3 distribution. RESULTS AND DISCUSSION: We found that AntiOxCIN3 has the potential to permeate the gastrointestinal tract. However, its biodistribution and elimination from the body might be affected by its affinity to HSA (>90%) and by its steady-state volume of distribution (VDSS =1.89± 0.48 L∙Kg-1). AntiOxCIN3 is expected to locate parallel to the membrane phospholipids, causing a bilayer stiffness effect. AntiOxCIN3 is also predicted to permeate through blood-brain barrier and reach its therapeutic target - the brain. CONCLUSION: Drug interactions with biological interfaces may be evaluated using membrane model systems and serum proteins. This knowledge is important for the characterization of drug partitioning, positioning and orientation of drugs in membranes, their effect on membrane biophysical properties and the study of serum protein binding. The analysis of these interactions makes it possible to collect valuable knowledge on the transport, distribution, accumulation and, eventually, therapeutic impact of drugs which may aid the drug development process.

All-Dielectric Synthetic-Phase Metasurfaces Generating Practical Airy Beams.

Accelerating optical beams exhibit exotic features, such as nondiffractive propagation, self-acceleration, and self-healing, which have led their use in a wide range of photonics applications. However, spatial light modulator-based generators of such beams suffer from narrow operational bandwidth, high cost, low diffraction efficiency, and limited integration capability. Although recent metasurface-based approaches have yielded generators with significantly improved bandwidths and integration capacities, the resultant devices usually have ultrashort working distances and limited control over characteristic beam parameters, which decreases their utility in optical imaging and manipulation applications. Herein, we describe a synthetic-phase metasurface-based approach that overcomes these problems and increases the degrees of freedom to enable effective control of beam parameters by integrating a cubic phase profile and the phase of a Fresnel holographic lens into a single metasurface. We demonstrate this approach by using the synthetic metasurface to generate a series of Airy beams with controllable focal length (i.e., working distance), narrowed beam width, and extended propagation distance. Crucially, these beam parameters are fully adjustable, which makes these focal-length-modifiable Airy beams particularly appealing for use in high-resolution, large field-of-view imaging, and deep-penetration optical manipulation. Furthermore, we show that imposing the phase of a Dammann grating into a synthetic metasurface generates a 1 × 4 array of Airy beams that exhibit the aforementioned optical properties. These findings suggest that synthetic-phase metasurfaces may significantly broaden the application of accelerating optical beams in various fields, such as light-sheet microscopy, super-resolution stochastic optical-reconstruction microscopy, laser fabrication, and parallel processing and in the development of optical tweezers for use with live samples.

Efficient light extraction in subwavelength GaAs/AlGaAs nanopillars for nanoscale light-emitting devices.

This work reports on high extraction efficiency in subwavelength GaAs/AlGaAs semiconductor nanopillars. We achieve up to 37-fold enhancement of the photoluminescence (PL) intensity from sub-micrometer (sub-µm) pillars without requiring back reflectors, high-Q dielectric cavities, nor large 2D arrays or plasmonic effects. This is a result of a large extraction efficiency for nanopillars <500 nm width, estimated in the range of 33-57%, which is much larger than the typical low efficiency (∼2%) of micrometer pillars limited by total internal reflection. Time-resolved PL measurements allow us to estimate the nonradiative surface recombination of fabricated pillars. We conclusively show that vertical-emitting nanopillar-based LEDs, in the best case scenario of both reduced surface recombination and efficient light out-coupling, have the potential to achieve notable large external quantum efficiency (∼45%), whereas the efficiency of large µm-pillar planar LEDs, without further methods, saturates at ∼2%. These results offer a versatile method of light management in nanostructures with prospects to improve the performance of optoelectronic devices including nanoscale LEDs, nanolasers, single photon sources, photodetectors, and solar cells.

Mapping intracellular thermal response of cancer cells to magnetic hyperthermia treatment.

Temperature is a key parameter for optimal cellular function and growth. Temperature perturbation may directly lead to cell death. This can be used in cancer therapies to kill cells in tumors, a therapeutic approach called hyperthermia. To avoid overheating of tumors that may damage healthy tissues, a knowledge of the intracellular temperature reached during the hyperthermia treatment of cancer cells is relevant. Recently, several luminescent intracellular nanothermometers have been proposed; however an application to sense temperature during a hyperthermia treatment is lacking. Here we present a technique to measure intracellular temperature changes in in vitro cancer cell models. For this purpose, we study for the first time the temperature dependence of the green fluorescent protein (GFP)'s fluorescence lifetime parameter. We find the fluorescence lifetime of GFP can be used for nanothermosensing. We use GFP in a bound form to actin filaments as an intracellular thermal reporter. Furthermore, we assess intracellular temperature during in vitro magnetothermal therapy on live HeLa cells incubated with polyacrylic acid-coated iron oxide nanoparticles. Compared to other thermosensitive materials and formulations reported so far, the GFP nanothermosensor is easily expressed via transfection and various GFP variants are commercially available. We foresee that the nanothermometer developed might find widespread applications in cancer therapy research and development.

Multi-beam two-photon polymerization for fast large area 3D periodic structure fabrication for bioapplications.

Two-photon polymerization (TPP) is capable of fabricating 3D structures with dimensions from sub-µm to a few hundred µm. As a direct laser writing (DLW) process, fabrication time of 3D TPP structures scale with the third order, limiting its use in large volume fabrication. Here, we report on a scalable fabrication method that cuts fabrication time to a fraction. A parallelized 9 multi-beamlets DLW process, created by a fixed diffraction optical element (DOE) and subsequent stitching are used to fabricate large periodic high aspect ratio 3D microstructured arrays with sub-micron features spanning several hundred of µm2. The wall structure in the array is designed with a minimum of traced lines and is created by a low numerical aperture (NA) microscope objective, leading to self-supporting lines omitting the need for line-hatching. The fabricated periodic arrays are applied in a cell - 3D microstructure interaction study using living HeLa cells. First indications of increased cell proliferation in the presence of 3D microstructures compared to planar surfaces are obtained. Furthermore, the cells adopt an elongated morphology when attached to the 3D microstructured surfaces. Both results constitute promising findings rendering the 3D microstructures a suited tool for cell interaction experiments, e.g. for cell migration, separation or even tissue engineering studies.

Prediction of paclitaxel pharmacokinetic based on in vitro studies: Interaction with membrane models and human serum albumin.

Interactions of paclitaxel (PTX) with models mimicking biological interfaces (lipid membranes and serum albumin, HSA) were investigated to test the hypothesis that the set of in vitro assays proposed can be used to predict some aspects of drug pharmacokinetics (PK). PTX membrane partitioning was studied by derivative spectrophotometry; PTX effect on membrane biophysics was evaluated by dynamic light scattering, fluorescence anisotropy, atomic force microscopy and synchrotron small/wide-angle X-ray scattering; PTX distribution/molecular orientation in membranes was assessed by steady-state/time-resolved fluorescence and computer simulations. PTX binding to HSA was studied by fluorescence quenching, derivative spectrophotometry and dynamic/electrophoretic light scattering. PTX high membrane partitioning is consistent with its efficacy crossing cellular membranes and its off-target distribution. PTX is closely located in the membrane phospholipids headgroups, also interacting with the hydrophobic chains, and causes a major distortion of the alignment of the membrane phospholipids, which, together with its fluidizing effect, justifies some of its cellular toxic effects. PTX binds strongly to HSA, which is consistent with its reduced distribution in target tissues and toxicity by bioaccumulation. In conclusion, the described set of biomimetic models and techniques has the potential for early prediction of PK issues, alerting for the required drug optimizations, potentially minimizing the number of animal tests used in the drug development process.

Phasor-assisted nanoscopy reveals differences in the spatial organization of major nuclear lamina proteins.

Phasor-assisted Metal Induced Energy Transfer-Fluorescence Lifetime Imaging Microscopy (MIET-FLIM) nanoscopy is introduced as a powerful tool for functional cell biology research. Thin metal substrates can be used to obtain axial super-resolution via nanoscale distance-dependent MIET from fluorescent dyes towards a nearby metal layer, thereby creating fluorescence lifetime contrast between dyes located at different nanoscale distance from the metal. Such data can be used to achieve axially super-resolved microscopy images, a process known as MIET-FLIM nanoscopy. Suitability of the phasor approach in MIET-FLIM nanoscopy is first demonstrated using nanopatterned substrates, and furthermore applied to characterize the distance distribution of the epithelial basal membrane of a biological cell from the gold substrate. The phasor plot of an entire cell can be used to characterize the full Förster resonance energy transfer (FRET) trajectory as a large distance heterogeneity within the sensing range of about 100 nm from the metal surface is present due to the extended shape of cell with curvatures. In contrast, the different proteins of nuclear lamina show strong confinement close to the nuclear envelope in nanoscale. We find the lamin B layer resides in average at shorter distances from the gold surface compared to the lamin A/C layer located in more extended ranges. This and the observed heterogeneity of the protein layer thicknesses suggests that A- and B-type lamins form distinct networks in the nuclear lamina. Our results provide detailed insights for the study of the different roles of lamin proteins in chromatin tethering and nuclear mechanics.

SyncRGB-FLIM: synchronous fluorescence imaging of red, green and blue dyes enabled by ultra-broadband few-cycle laser excitation and fluorescence lifetime detection.

We demonstrate for the first time that an ultra-broadband 7 femtosecond (fs) few-cycle laser can be used for multicolor nonlinear imaging in a single channel detection geometry, when employing a time-resolved fluorescence detection scheme. On a multi-chromophore-labelled cell sample we show that the few-cycle laser can efficiently excite the multiple chromophores over a >400 nm two-photon absorption range. By combining the few-cycle laser excitation with time-correlated single-photon counting (TCSPC) detection to record two-photon fluorescence lifetime imaging microscopy (FLIM) images, the localization of different chromophores in the cell can be identified based on their fluorescence decay properties. The novel SyncRGB-FLIM multi-color bioimaging technique opens the possibility of real-time protein-protein interaction studies, where its single-scan operation translates into reduced laser exposure of the sample, resulting in more photoprotective conditions for biological specimens.

GFP fluorescence peak fraction analysis based nanothermometer for the assessment of exothermal mitochondria activity in live cells.

Nanothermometry methods with intracellular sensitivities have the potential to make important contributions to fundamental cell biology and medical fields, as temperature is a relevant physical parameter for molecular reactions to occur inside the cells and changes of local temperature are well identified therapeutic strategies. Here we show how the GFP can be used to assess temperature-based on a novel fluorescence peak fraction method. Further, we use standard GFP transfection reagents to assess temperature intracellularly in HeLa cells expressing GFP in the mitochondria. High thermal resolution and sensitivity of around 0.26% °C-1 and 2.5% °C-1, were achieved for wt-GFP in solution and emGFP-Mito within the cell, respectively. We demonstrate that the GFP-based nanothermometer is suited to directly follow the temperature changes induced by a chemical uncoupler reagent that acts on the mitochondria. The spatial resolution allows distinguishing local heating variations within the different cellular compartments. Our discovery may lead to establishing intracellular nanothermometry as a standard method applicable to the wide range of live cells able to express GFP.

Surface charge tunable catanionic vesicles based on serine-derived surfactants as efficient nanocarriers for the delivery of the anticancer drug doxorubicin.

Self-assembled vesicles composed of amino acid-based cationic/anionic surfactant mixtures show promise as novel effective drug nanocarriers. Here, we report the in vitro performance of vesicles based on cationic (16Ser) and anionic (8-8Ser) serine-based surfactants using a cancer cell model for the delivery of the anticancer drug doxorubicin (DOX). This catanionic mixture yields both negatively (0.20 in the cationic surfactant molar fraction, x16Ser) and positively (x16Ser = 0.58) charged vesicles, hence providing a surface charge tunable system. Low toxicity is confirmed for concentration ranges below 32 µM in both formulations. DOX is successfully encapsulated in the vesicles, resulting in a surface charge switch to negative for the (0.58) system, making both (0.20) and (0.58) DOX-loaded vesicles highly interesting for systemic administration. High uptake by cells was demonstrated using flow cytometry and confocal microscopy. Drug accumulation results in an increase of cell uptake up to 250% and 200% for the (0.20) and (0.58) vesicles, respectively, compared to free DOX and with localizations near the nuclear regions in the cells. The in vitro cytotoxicity studies show that DOX-loaded vesicles induce cell death, confirming the therapeutic potential of the formulations. Furthermore, the efficient accumulation of the drug inside the cell compartments harbors the potential for optimization strategies including phased delivery for prolonged treatment periods or even on-demand release.