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Myocardial remodeling is an inevitable risk factor for cardiac arrhythmias and can potentially be corrected with cell therapy. Although the generation of cardiac cells ex vivo is possible, specific approaches to cell replacement therapy remain unclear. On the one hand, adhesive myocyte cells must be viable and conjugated with the electromechanical syncytium of the recipient tissue, which is unattainable without an external scaffold substrate. On the other hand, the outer scaffold may hinder cell delivery, for example, making intramyocardial injection difficult. To resolve this contradiction, we developed molecular vehicles that combine a wrapped (rather than outer) polymer scaffold that is enveloped by the cell and provides excitability restoration (lost when cells were harvested) before engraftment. It also provides a coating with human fibronectin, which initiates the process of graft adhesion into the recipient tissue and can carry fluorescent markers for the external control of the non-invasive cell position. In this work, we used a type of scaffold that allowed us to use the advantages of a scaffold-free cell suspension for cell delivery. Fragmented nanofibers (0.85 µm ± 0.18 µm in diameter) with fluorescent labels were used, with solitary cells seeded on them. Cell implantation experiments were performed in vivo. The proposed molecular vehicles made it possible to establish rapid (30 min) electromechanical contact between excitable grafts and the recipient heart. Excitable grafts were visualized with optical mapping on a rat heart with Langendorff perfusion at a 0.72 ± 0.32 Hz heart rate. Thus, the pre-restored grafts’ excitability (with the help of a wrapped polymer scaffold) allowed rapid electromechanical coupling with the recipient tissue. This information could provide a basis for the reduction of engraftment arrhythmias in the first days after cell therapy.

The main limitation of the use of tip-enhanced Raman scattering (TERS) is due to the lack of reliable scanning probes. We present a simple procedure to manufacture TERS probes with highly reproducible characteristics that are reliable and provide sufficient enhancement. The procedure is based on the modification of conventional cantilevers of scanning probe microscopes by plasma etching, followed by the formation of a TERS enhancing region at the apex by deposition of colloidal nanoparticles by dielectrophoresis.

A unified analytical technology based on the integration of scanning probe nanotomography and optical microscopy (SPNT–OM) is presented, and the potential of the technology is demonstrated by the example of a multiparameter analysis of encoded microspheres. The presented SPNT–OM technology can become a powerful tool for multimodal nanocharacterization of a wide range of microparticles, composites, and hybrid polymers.

A comparative analysis of the structure and biological properties of silk fibroin constructions was performed. Three groups of constructions were obtained: films obtained by casting an aqueous solution of silk fibroin and electrospun microfibrous scaffolds based on silk fibroin, with the addition of 30% gelatin per total protein weight. The internal structures of the films and single fibers of the microfibrous scaffolds consisted of densely packed globule structures, the surface area to volume ratios and volume porosities of the microfibrous scaffolds were calculated. All constructions were non-toxic for cells and provide high levels of adhesion and proliferation. The high regenerative potential of the constructions was demonstrated in a rat full-thickness skin wound healing model. The constructions accelerated healing by an average of 15 days and can be considered to be promising constructions for various tasks of tissue engineering and regenerative medicine.

We present a 3D study of nanostructural features of a bioprinted tissue spheroid interacting with polyurethane dual-scale biocompatible scaffold manufactured by three-dimensional printing and electrospinning. Three-dimensional analysis of fibroblasts interacting with electrospun polyurethane fibers was conducted using scanning probe nanotomography with an experimental setup combining ultramicrotome and a scanning probe microscope. Three-dimensional reconstruction demonstrates direct visualization of cell membrane protrusions and coherent cell-fiber interfaces, the formation of which is a prerequisite for an efficient tissue engineered implant. Analysis of obtained 3D data allows for quantitative calculation of the important morphological parameters of adhered cells, scaffolds, and cell-scaffold interfaces. The proposed method may be successfully applied to investigate 3D cell-scaffold constructs at nanoscale.

Organ decellularization is one of the promising technologies of regenerative medicine, which allows obtaining cell-free extracellular matrix (ECM), which provide preservation of the composition, architecture, vascular network and biological activity of the ECM. The method of decellularization opens up wide prospects for its practical application not only in the field of creating full-scale bioengineered structures, but also in the manufacture of vessels, microcarriers, hydrogels, and coatings. The main goal of our work was the investigation of structure and biological properties of lyophilized decellularized Wistar rat liver fragments (LDLFs), as well as we assessed the regenerative potential of the obtained ECM. We obtained decellularized liver of a Wistar rat, the vascular network and the main components of the ECM of tissue were preserved. H&E staining of histological sections confirmed the removal of cells. DNA content of ECM is equal to 0.7% of native tissue DNA content. Utilizing scanning probe nanotomogrphy method, we showed sinuous, rough topography and highly nanoporous structure of ECM, which provide high level of mouse 3T3 fibroblast and Hep-G2 cells biocompatibility. Obtained LDLF had a high regenerative potential, which we studied in an experimental model of a full-thickness rat skin wound healing: we observed the acceleration of wound healing by 2.2 times in comparison with the control.

The layer-by-layer (LbL) deposition approach allows combined incorporation of fluorescent, magnetic, and plasmonic nanoparticles into the shell of polyelectrolyte microcapsules to obtain stimulus-responsive systems whose imaging and drug release functions can be triggered by external stimuli. The combined use of fluorescent quantum dots (QDs) and magnetic nanoparticles (MNPs) yields magnetic-field-driven imaging tools that can be tracked and imaged even deep in tissue when the appropriate type of QDs and wavelength of their excitation are used. QDs are excellent photonic labels for microcapsule encoding due to their close-to-unity photoluminescence (PL) quantum yields, narrow PL emission bands, and tremendous one- and two-photon extinction coefficients. However, the presence of MNPs and electrically charged polyelectrolyte molecules used for the LbL fabrication of magneto-optical microcapsules provokes alterations of the QD optical properties because of the photoinduced charge and energy transfer resulting in QD photodarkening or photobrightening. These lead to variation of the microcapsule PL signal under illumination, which hampers their tracking and quantitative analysis in cells and tissues. Here, we have studied the effects of the structure and spatial arrangement of the nanoparticles within the microcapsule polyelectrolyte shell, the total shell thickness, and the shell surface charge on their PL properties under continuous illumination. The roles of the charge transfer and its main driving forces in the stability of the microcapsules PL signal have been established, and the design of the microcapsules dually encoded with QDs and MNPs providing the strongest and most stable PL has been determined. Controlling the energy transfer from the QDs and MNPs and the charge transfer from QDs to polyelectrolyte layers in the engineering of magneto-optical microcapsules with a bright and stable PL signal extends their applications to long-lasting quantitative fluorescence imaging.

Building functional and robust scaffolds for engineered biological tissue requires a nanoscale mechanistic understanding of how cells use the scaffold for their growth and development. A vast majority of the scaffolds used for cardiac tissue engineering are based on polymer materials, the matrices of nanofibers. Attempts to load the polymer fibers of the scaffold with additional sophisticated features, such as electrical conductivity and controlled release of the growth factors or other biologically active molecules, as well as trying to match the mechanical features of the scaffold to those of the extracellular matrix, cannot be efficient without a detailed knowledge of how the cells are attached and strategically positioned with respect to the scaffold nanofibers at micro and nanolevel. Studying single cell – single fiber interactions with the aid of confocal laser scanning microscopy (CLSM), scanning probe nanotomography (SPNT), and transmission electron microscopy (TEM), we found that cardiac cells actively interact with substrate nanofibers, but in different ways. While cardiomyocytes often create a remarkable “sheath” structure, enveloping fiber and, thus, substantially increasing contact zone, fibroblasts interact with nanofibers in the locations of focal adhesion clusters mainly without wrapping the fiber. Statements of Significance We found that cardiomyocytes grown on electrospun polymer nanofibers often create a striking “sheath” structure, enveloping fiber with the formation of a very narrow (∼22 nm) membrane gap leading from the fiber to the extracellular space. This wrapping makes the entire fiber surface available for cell attachment. This finding gives a new prospective view on how scaffold nanofibers may interact with growing cells. It may play a significant role in effective design of novel nanofiber scaffolds for tissue engineering concerning mechanical and electrical properties of scaffolds as well as controlled drug release from “smart” biomaterials.

In the past decade correlative microscopy, which combines the potentials of different types of high-resolution microscopies with a variety of optical microspectroscopy techniques, has been attracting increasing attention in material science and biological research. One of outstanding solutions in this area is the combination of scanning probe microscopy (SPM), which provides data on not only the topography, but also the spatial distribution of a wide range of physical properties (elasticity, conductivity, etc.), with ultramicrotomy, allowing 3D multiparametric examination of materials. The combination of SPM and ultramicrotomy (scanning probe nanotomography) is very appropriate for characterization of soft multicompound nanostructurized materials, such as polymer matrices and microstructures doped with different types of nanoparticles (magnetic nanoparticles, quantum dots, nanotubes, etc.), and biological materials. A serious problem of this technique is a lack of chemical and optical characterization tools, which may be solved by using optical microspectroscopy. Here, we report the development of an instrumental approach to combining confocal microspectroscopy and 3D scanning probe nanotomography in a single apparatus. This approach retains all the advantages of SPM and upright optical microspectroscopy and allows 3D multiparametric characterization using both techniques. As the first test of the system developed, we have performed correlative characterization of the morphology and the magnetic and fluorescent properties of fluorescent magnetic microspheres doped with a fluorescent dye and magnetic nanoparticles. The results of this study can be used to obtain 3D volume images of a specimen for most high-resolution near-field scanning probe microscopies: SNOM, TERS, AFM-IR, etc. This approach will result in development of unique techniques combining the advantages of SPM (nanoscale morphology and a wide range of physical parameters) and high-resolution optical microspectroscopy (nanoscale chemical mapping and optical properties) and allowing simultaneous 3D measurements.

Tissue-specific alginate microspheres surface modified with microparticles of decellularized liver tissue were prepared. The nanostructures of these microspheres were analysed by cryo scanning probe nanotomography. Modification of the surfaces of the alginate microspheres with microparticles of decellularized liver tissue increased surface roughness, the specific effective area, and the average autocorrelation length. Up to seven times more human hepatocellular carcinoma (HepG2) cells were adhered to the modified alginate microspheres than to the unmodified microspheres. The developed alginate microspheres can be used in regenerative medicine and tissue engineering as a biodegradable cell microcarriers.

International audience; Fluorescent imaging is a widely used technique for detecting and monitoring the distribution, interaction, and transformation processes at molecular, cellular, and tissue level in modern diagnostic and other biomedical applications. Unique photophysical properties of fluorescent semiconductor nanocrystals "quantum dots" (QDs) make them advanced fluorophores for fluorescent labeling of biomolecules or optical encoding of microparticles to be used as bioimaging and theranostic agents in targeted delivery, visualization, diagnostics, and imaging. This paper reports on the results of development of an improved approach to the optical encoding of polyelectrolyte microcapsules with stable, covered with the multifunctional polyethyleneglycol derivatives water-soluble QDs, as well as characterization of the optical properties, morphological and structural properties of the encoded microcapsules. The embedding of QDs into the polymer microcapsule membrane through layer-by-layer deposition on a preliminarily formed polymeric polyelectrolyte shell makes it possible to obtain bright fluorescent particles with an adapted charge and size distribution that are distinctly discernible by flow cytometry as individual homogeneous populations. The fluorescent microcapsules developed can be used in further designing bioimaging and theranostic agents sensitive to various external stimuli along with photoexcitation.

This paper presents a study of nanostructural features of contacts of bioprinted tissue spheroids with polyurethane dual scale biocompatible scaffold made by three-dimensional printing and electrospinning. Analysis of nanostructural features of cell contacts was carryed out by scanning probe microscopy with use of experimental setup combining ultramicrotome and scanning probe microscope. Measured mean cell volume is 460 ± 104 μm3, mean contact area of cells with scaffold fibers–104.8 μm2 per cell (16.7% of total cell area). Maximum distance of migrating cells from spheroid border at 48 h. is ~200 μm, what corresponds to mean velocity of cell migration more than 4 μm/h. Obtained quantitative characteristics of micro- and nanostructure of human fibroblast cell contacts with elecrospun polyurethane scaffold secure high efficacy of tissue regeneration with its usage for implanted bioprinted dual scale tissue-engineered scaffolds.

This paper presents a study of three-dimensional micro- and nanosctucture of polyurethane dual scale biocompatible scaffold made by three-dimensional printing and electrospinning. The three-dimensional structure of the scaffold was analyzed by scanning probe nanotomography with use of an experimental setup combining an ultramicrotome and a scanning probe microscope. We performed a quantitative analysis of microporosity, nanoroughness, and three-dimensional morphology parameters of the scaffold. The electrospun scaffold consists of a network of microfibers with diameter ranging from 1.7 to 6.0 μm. The measured mean microfiber diameter is 3.54 ± 1.23 μm. The volume porosity of the electrospun scaffold is 72.5%, while mean surface area to volume ration is 0.28 μm–1 and mean nanoroughness of microfiber surface is 22.1 ± 3.0 nm. The quantitative characteristics of the micro- and nanostructure of elecrospun polyurethane matrices secure the high efficacy of its usage for increasing the biocompatibility of dual-scale hybrid bioengineered scaffolds for regenerative medicine tasks. The use of scanning probe nanotomography for analyzing threedimensional morphology characteristics and the topology of electrospun microfiber systems enables us to improve the efficiency of development of new bioengineered products.

International audience; Quantum dot (QD) encoded microbeads are emerging for multiplexed analysis of biological markers. The quantitative encoding of microbeads prepared with different concentrations of QDs of different colors suffers from resonance energy transfer from the QDs fluorescing at shorter wavelengths to the QDs fluorescing at longer wavelengths. Here, we used the layer‐by‐layer deposition technique to spatially separate QDs of different colors with several polymer layers so that the distance between them would be larger than the Förster energy transfer radius. We performed fluorescence lifetime measurements to investigate and determine the conditions excluding significant resonance energy transfer between QDs within QD‐encoded microbeads. Additionally, the number of QDs adsorbed onto microbeads was systematically established and multilayer structures of the QD‐encoded microbead shells were characterized by scanning probe nanotomography. Finally, we prepared eight populations of FRET‐free microbeads encoded with QDs of three colors at two intensity levels and demonstrated that all the optical codes are excitable at a single wavelength and may be clearly identified in three channels of a flow cytometer. The developed approach for engineering QD‐encoded microbeads that are free from optical artefacts related to inter‐QD resonance energy transfer paves the way to quantitative QD‐based multiplexed assays.

We use serial section scanning probe nanotomography to study the 3D structure, nanoporosity and pore interconnectivity of biocompatible scaffolds made of fibroin and recombinant spidroin 1 (rS1/9). Significant nanoporosity (24%) and pore percolation is detected for rS1/9 scaffolds. It is assumed to contribute to higher in vivo tissue regeneration efficiency of rS1/9 scaffolds.

This paper presents a study of three-dimensional micro- and nanosctuctures of porous biocompatible matrices from regenerated fibroin and a quantitative analysis of their microporosity parameters. An analysis of the three-dimensional structure of matrices has been carried out by scanning probe nanotomography with the use of an experimental setup combining an ultramicrotome and scanning probe microscope. The formation of a three-dimensional network of interconnected pores with characteristic dimensions ranging from 1.7 to 6.0 μm is observed in the bulk volume of studied matrices. The measured mean pore diameter is 3.54 ± 1.23 μm; the mean pore wall thickness is 672 ± 282 nm. The volume porosity of macropore walls is 65.7%, while the volume fraction of pores interconnected in large pore clusters is more than 80% of the whole pore volume. Quantitative characteristics of porous micro- and nanostructures of matrices obtained as a result of the study show a significant degree of porosity and percolation of micropores, which correlates with the reported high efficiency of tissue regeneration on such matrices. The use of scanning probe nanotomography to analyze three-dimensional morphology characteristics and the topology of micro- and nanopore systems enables us to improve the efficiency of developing new biomaterials.

Scanning Probe Tomography (SPT) is a new method for nanoscale volume imaging of sample morphology and property distribution based on Scanning Probe Microscopy (SPM). In this review we describe and discuss recent results obtained with different SPT techniques on polymer samples. The design of the existing SPM based instruments used for tomography in principle allows for volume reconstruction of any kind of mechanical, functional or chemical property distribution, which can be measured by SPM. We describe some recent volume reconstruction results from several types of polymer materials and critically discuss limitations and future prospects of SPM tomography.

International audience; A new experimental approach to multiparametric three-dimensional (3D) investigation of a broad class of composite nanostructural materials is developed on the basis of scanning near-field optical nanotomography (SNONT). Using this method, it is possible to simultaneously study the optical properties, 3D morphology, and distribution of the mechanical and electrical properties of the same region of a sample. The proposed method combines features of the confocal and near-field optical microspectroscopy (fluorescence and Raman spectroscopy) with a lateral resolution of up to 50 nm and scanning-probe microscopy. The possibility of studying the volume distribution of optical, morphological, electrical, and mechanical characteristics of a material with nanoscale resolution is related to the probing of sequential layers at a step of up to 20 nm and a total Z-scan depth of up to 3 mm. In particular, the SNONT method has been used to study a liquid-crystalline polymer doped with fluorescent nanocrystals.

Self-sensing and –actuating probes optimized for conventional tapping mode atomic force microscopy (AFM) are described. 32-kHz quartz tuning forks with a chemically etched and focus ion beam (FIB) sharpened (curvature radii are 5-10 nm) tungsten tip are stable at air and liquid nitrogen atmosphere and at a wide range of temperatures. If driven at constant frequency, the scan speed of such sensors can be up to 3 Hz. AFM was performed on polymer samples in order to study the stability and applicability of these sensor for investigation of soft materials with high dynamical tendencies.

The correlative microscopy method based on a combination of optical and electronic techniques that is increasingly widely used now, has a number of limitations. Here, an alternative approach is considered that uses scanning probe microscopy (SPM) technique to get high-resolution and ultra-high-resolution data. SPM greatly increases the possibilities of collecting new information (on topological, morphological, electrical, magnetic etc. properties). To obtain three-dimensional distributions of different parameters of the sample, ultramicrotomography is used, which allows to scan the sample in steps of up to 20 nm. The principal advantage of the approach is that spectral data are used which due to the combination in near field microscopy can be gained with high and ultrahigh resolution. All above mentioned features are implemented in a single instrument, which allows to have 3-D data and their distributions at the same instrumental platform. A special feature of the approach is the possibility to use all the power of micro(nano)spectral methods. Therefore, it would be more correct to name the proposed approach ”Correlative microspectroscopy”. Keywords: correlative microscopy, correlative microspectroscopy, scanning near-ielf optical microscopy, ultramicrotomography, Raman, TERS

A new device (NTEGRA Tomo) that is based on the integration of the scanning probe microscope (SPM) (NT-MDT NTEGRA SPM) and the Ultramicrotome (Leica UC6NT) is presented. This integration enables the direct monitoring of a block face surface immediately following each sectioning cycle of ultramicrotome sectioning procedure. Consequently, this device can be applied for a serial section tomography of the wide range of biological and polymer materials. The automation of the sectioning/scanning cycle allows one to acquire up to 10 consecutive sectioned layer images per hour. It also permits to build a 3-D nanotomography image reconstructed from several tens of layer images within one measurement session. The thickness of the layers can be varied from 20 to 2000 nm, and can be controlled directly by its interference colour in water. Additionally, the NTEGRA Tomo with its nanometer resolution is a valid instrument narrowing and highlighting an area of special interest within volume of the sample. For embedded biological objects the ultimate resolution of SPM mostly depends on the quality of macromolecular preservation of the biomaterial during sample preparation procedure. For most polymer materials it is comparable to transmission electron microscopy (TEM). The NTEGRA Tomo can routinely collect complementary AFM and TEM images. The block face of biological or polymer sample is investigated by AFM, whereas the last ultrathin section is analyzed with TEM after a staining procedure. Using the combination of both of these ultrastructural methods for the analysis of the same particular organelle or polymer constituent leads to a breakthrough in AFM/TEM image interpretation. Finally, new complementary aspects of the object's ultrastructure can be revealed.

An immunodiagnostic lab-on-a-bead suspension microarray based on microbeads encoded with quantum dots (QDs) has been developed and preclinically validated for multiplexed quantitative detection of prostate cancer markers in human serum samples. The sensitivity and specificity of the microarray are similar to those of "gold-standard" single-analyte ELISA. Moreover, the array has an improved immunoassay capacity, ensures quantitative detection of multiple cancer biomarkers and may be operational in a considerably wider dynamic range of concentrations. The array is characterized by reduced time and cost of analysis and is compatible with classical flow cytometers. Proof-of-concept preclinical tests ensured simultaneous quantitative determination of free and total prostate-specific antigens in human serum, with clear discrimination between the control and clinical samples. The proposed approach is flexible and paves the way to development of a wide variety of immunodiagnostic assays for multiplexed early diagnosis of various diseases. From the Clinical Editor Early diagnosis of cancer can result in better prognosis for patients. Thus, the use of specific tumor markers is widely employed in clinical practice. Traditional screening methods only employ single markers. The authors here developed a microarray system based on microbeads encoded with quantum dots (QDs), which can be used for multiplexed quantitative detection. The validated results on patient samples should lead to the development of a wider variety of assays for other diseases.

International audience; An instrumental system and the corresponding experimental technique for multiparametric nanoscale 3D characterization of a wide range of composite nanomaterials have been developed. This system makes it possible to obtain 3D data on the chemical composition of a material by optical methods in the confocal and near-field optical microspectroscopy modes (fluorescence and Raman) with a lateral resolution up to 50 nm, as well as data on the 3D morphology and spatial distribution of mechanical, electrical, and other characteristics of the material in the scanning probe microscopy mode with a resolution of about 10 nm on the X and Y axes and several angstroms on the Z axis, for a single area of the sample (100 μm × 100 μm × 3 mm). The nanoscale 3D pattern of the distribution of these characteristics is obtained by sequentially examining nanomaterial layers at a step of 20 nm along the Z axis and a total depth of Z-scanning of 3 mm.

International audience; We present a new concept of a combined scanning probe microscope (SPM)/ultramicrotome apparatus. It enables “slice-and-view” scanning probe nanotomography measurements and 3D reconstruction of the bulk sample nanostructure from series of SPM images after consecutive ultrathin sections. The sample is fixed on a flat XYZ scanning piezostage mounted on the ultramicrotome arm. The SPM measuring head with a cantilever tip and a laser-photodiode tip detection system approaches the sample for SPM measurements of the block-face surface immediately after the ultramicrotome sectioning is performed. The SPM head is moved along guides that are also fixed on the ultramicrotome arm. Thereby, relative dysfunctional displacements of the tip, the sample, and the ultramicrotome knife are minimized. The design of the SPM head enables open frontal optical access to the sample block-face adapted for high-resolution optical lenses for correlative SPM/optical microscopy applications. The new system can be used in a wide range of applications for the study of 3D nanostructures of biological objects, biomaterials, polymer nanocomposites, and nanohybrid materials in various SPM and optical microscopy measuring modes.

International audience; Advanced multiplexed assays have recently become an indispensable tool for clinical diagnostics. These techniques provide simultaneous quantitative determination of multiple biomolecules in a single sample quickly and accurately. The development of multiplex suspension arrays is currently of particular interest for clinical applications. Optical encoding of microparticles is the most available and easy-to-use technique. This technology uses fluorophores incorporated into microbeads to obtain individual optical codes. Fluorophore-encoded beads can be rapidly analyzed using classical flow cytometry or microfluidic techniques. We have developed a new generation of highly sensitive and specific diagnostic systems for detection of cancer antigens in human serum samples based on microbeads encoded with fluorescent quantum dots (QDs). The designed suspension microarray system was validated for quantitative detection of (1) free and total prostate specific antigen (PSA) in the serum of patients with prostate cancer and (2) carcinoembryonic antigen (CEA) and cancer antigen 15-3 (CA 15-3) in the serum of patients with breast cancer. The serum samples from healthy donors were used as a control. The antigen detection is based on the formation of an immune complex of a specific capture antibody (Ab), a target antigen (Ag), and a detector Ab on the surface of the encoded particles. The capture Ab is bound to the polymer shell of microbeads via an adapter molecule, for example, protein A. Protein A binds a monoclonal Ab in a highly oriented manner due to specific interaction with the Fc-region of the Ab molecule. Each antigen can be recognized and detected due to a specific microbead population carrying the unique fluorescent code. 100 and 231 serum samples from patients with different stages of prostate cancer and breast cancer, respectively, and those from healthy donors were examined using the designed suspension system. The data were validated by comparing with the results of the “gold standard” enzyme-linked immunosorbent assay (ELISA). They have shown that our approach is a good alternative to the diagnostics of cancer markers using conventional assays, especially in early diagnostic applications.

International audience; Combination of 3D structural analysis with optical characterization of the same sample area on the nanoscale is a highly demanded approach in nanophotonics, materials science, and quality control of nanomaterial. We have developed a correlative microscopy technique where the 3D structure of the sample is reconstructed on the nanoscale by means of a “slice-and-view” combination of ultramicrotomy and scanning probe microscopy (scanning probe nanotomography, SPNT), and its optical characteristics are analyzed using microspectroscopy. This approach has been used to determine the direct quantitative relationship of the 3D structural characteristics of nanovolumes of materials with their microscopic optical properties. This technique has been applied to 3D structural and optical characterization of a hybrid material consisting of cholesteric liquid crystals doped with fluorescent quantum dots (QDs) that can be used for photochemical patterning and image recording through the changes in the dissymmetry factor of the circular polarization of QD emission. The differences in the polarization images and fluorescent spectra of this hybrid material have proved to be correlated with the arrangement of the areas of homogeneous distribution and heterogeneous clustering of QDs. The reconstruction of the 3D nanostructure of the liquid crystal matrix in the areas of homogeneous QDs distribution has shown that QDs do not perturb the periodic planar texture of the cholesteric liquid crystal matrix, whereas QD clusters do perturb it. The combined microspectroscopy–nanotomography technique will be important for evaluating the effects of nanoparticles on the structural organization of organic and liquid crystal matrices and biomedical materials, as well as quality control of nanotechnology fabrication processes and products.

International audience; Fabrication of modern nanomaterials and nanostructures with specific functional properties is both scientifically promising and commercially profitable. The preparation and use of nanomaterials require adequate methods for the control and characterization of their size, shape, chemical composition, crystalline structure, energy levels, pathways and dynamics of physical and chemical processes during their fabrication and further use. In this review, we discuss different instrumental methods for the analysis and metrology of materials and evaluate their advantages and limitations at the nanolevel.

Combination of nanometer-scale 3D structural analysis with optical characterization of the same material is a challenging task. Its results may be important for nanophotonics, materials science, and quality control. We have developed a new technique for complementary high-resolution structural and optical characterization followed by optical spectroscopic and microscopic measurements accompanied by reconstruction of the 3D structure in the same area of the sample. The 3D structure is reconstructed by combination of ultramicrotomic and SPM techniques allowing the study of the 3D distribution of implanted nanoparticles and their effect on the matrix structure. The combination of scanning probe nanotomography (SPN) and optical microspectroscopy makes it possible to direct estimate how the 3D structural characteristics of materials affect their macroscopic optical properties. The technique developed has been applied to the engineering of materials made from cholesteric liquid crystals and fluorescent quantum dots (QDs). These materials permit photochemical patterning and image recording through the changes in the dissymmetry factor of circular polarization of QD emission. The differences in the polarisation images and morphological characteristics of the liquid crystal matrix have proved to be correlated with the arrangement of the areas of homogeneous distribution and nonhomogeneous clustering of QDs. The reconstruction of the 3D structure of the liquid crystal matrix in the areas of homogeneous QD distribution has shown that QDs embedded into cholesteric liquid crystal matrices do not perturb their periodic planar texture. The combined optical/SPM/ultramicrotome technique will be indispensable for evaluating the effects of inorganic nanoparticles on the organisation of organic and liquid crystal matrices, biomedical materials, cells, and tissues.

We propose a universal tool for structural analysis of soft/frozen-hydrated materials at a macromolecular level over a wide range of temperatures (−120 to +50 °C). The cryogenic scanning probe microscope presented here allows detailed investigation and serial-section 3D tomography of soft matter under native-like conditions.

International audience; Optical materials based on cholesteric liquid crystals (LCs) doped with fluorescent CdSe/ZnS quantum dots (QDs) have been developed and demonstrated to have a wide photonic band gap. It has been shown that the fluorescence emission of QDs embedded in LCs is circularly polarized and that the dissymmetry factor of this polarization may be optically or electrically controlled via conformational changes in the helical structure of the LC matrix. The possibility of photochemical patterning or image recording using these materials has been demonstrated; the recorded information can be read through changes in the dissymmetry factor of circular polarization of QDs emission. The developed photo- and electro-active materials with a controlled degree of fluorescence circular polarization may be used as on-demand single photon sources in photonics, optoelectronics, and quantum cryptography, as well as for development of nanophotonic systems capable of low-threshold lasing.

Micro and nanostructure of scaffolds made from fibroin of Bombyx mori silkworm by salt leaching technique was studied by scanning probe nanotomography. Nanopores with dimensions in range from 30 to 180nm are observed in the scaffold volume. Three – dimensional analysis of obtained data shows that degree of scaffold nanoporosity is 0.5% and nanopores are not interconnected with each other. Usage of scanning probe nanotomography technique enables to obtain unique nanoscale information of 3D structure of biopolymer nanomaterials.