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1.
J Am Chem Soc ; 146(3): 2196-2207, 2024 Jan 24.
Artículo en Inglés | MEDLINE | ID: mdl-38214460

RESUMEN

Three incongruent melting LawNdxGdyYzSc4-w-x-y-z(BO3)4 (LGYSB:Nd) crystals with different Y concentrations (z = 0.15, 0.05, and 0.025) have been grown by the Czochralski method for the first time. The LGYSB:Nd-type crystals exhibit an acentric structure similar to that of the natural mineral huntite CaMg3(CO3)4, with space group R32. The composition of the LGYSB:Nd_3 (z = 0.025) crystal grown from the starting melt composition La0.628Gd0.547Y0.025Nd0.05Sc2.75(BO3)4 was measured by inductively coupled plasma mass spectrometry method, and it was found to be La0.6794Gd0.4105Y0.0178Nd0.0381Sc2.8542(BO3)4. The transversal spatial distribution of the refractive index in the LGYSB:Nd_3 crystal was investigated. Third-order nonlinear optical susceptibility χ(3) of the LGYSB:Nd_3 crystal was determined from third-harmonic generation experiments with ultrashort (fs) laser pulses. The optical transmission spectrum was measured in the range of 200-2000 nm. The absorption cross-section at 808 nm in σ-polarization was determined to be 1.18 × 10-19 cm2 for the LGYSB:Nd_3 crystal. The 10K absorption spectra revealed that the Nd3+ ions occupy only La3+ cationic sites in the LGYSB host matrix. The emission cross-section at 1064 nm in σ-polarization was determined to be σem(σ) = 1.74 × 10-19 cm2 for the LGYSB:Nd_3 crystal. The fluorescence lifetime was found to be τ = 115 µs for all of the LGYSB:Nd crystals. The LGYSB:Nd_3 laser was operated at an emission wavelength of 1062 nm with very high slope efficiencies of ηsa = 0.74 and ηsa = 0.64 in quasi-cw and cw regimes, respectively.

2.
RSC Adv ; 13(29): 19682-19694, 2023 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-37396836

RESUMEN

The fast and global spread of bacterial resistance to currently available antibiotics results in a great and urgent need for alternative antibacterial agents and therapeutic strategies. Recent studies on the application of nanomaterials as antimicrobial agents have demonstrated their potential for the management of infectious diseases. Among the diverse palette of nanomaterials currently used in biomedical applications, carbon nanotubes (CNTs) have gained massive interest given their many valuable properties, such as high thermal and electrical conductivity, tensile strength, flexibility convenient aspect ratio, and low fabrication costs. All these features are augmented by facile conjugation with functional groups. CNTs are currently available in many configurations, with two main categories being single-walled and multi-walled CNTs, depending on the number of rolled-up single-layer carbon atoms sheets making up the nanostructure. Both classes have been identified over the past years as promising antibacterial agents but the current level of understanding of their efficiency still harbors many pending questions. This mini-review surveys recent progress on the topic of antibacterial effects of CNTs and examines the proposed mechanisms of action(s) of different CNT typologies, placing the main focus on past studies addressing the antibacterial activity on Staphylococcus aureus and Escherichia coli, two prototypical Gram-positive and Gram-negative pathogens, respectively.

3.
Nanomaterials (Basel) ; 13(4)2023 Feb 04.
Artículo en Inglés | MEDLINE | ID: mdl-36838989

RESUMEN

In this study, we report on the synthesis of L-Cysteine (L-Cys)-coated magnetic iron oxide nanoparticles (NPs) loaded with doxorubicin (Dox). The Fe3O4-L-Cys-Dox NPs were extensively characterized for their compositional and morpho-structural features using EDS, SAED, XRD, FTIR and TEM. XPS, MÓ§ssbauer spectroscopy and SQUID measurements were also performed to determine the electronic and magnetic properties of the Fe3O4-L-Cys-Dox nanoparticles. Moreover, by means of a FO-SPR sensor, we evidenced and confirmed the binding of Dox to L-Cys. Biological tests on mouse (B16F10) and human (A375) metastatic melanoma cells evidenced the internalization of magnetic nanoparticles delivering Dox. Half maximum inhibitory concentration IC50 values of Fe3O4-L-Cys-Dox were determined for both cell lines: 4.26 µg/mL for A375 and 2.74 µg/mL for B16F10, as compared to 60.74 and 98.75 µg/mL, respectively, for unloaded controls. Incubation of cells with Fe3O4-L-Cys-Dox modulated MAPK signaling pathway activity 3 h post-treatment and produced cell cycle arrest and increased apoptosis by 48 h. We show that within the first 2 h of incubation in physiological (pH = 7.4) media, ~10-15 µM Dox/h was released from a 200 µg/mL Fe3O4-L-Cys-Dox solution, as compared to double upon incubation in citrate solution (pH = 3), which resembles acidic environment conditions. Our results highlight the potential of Fe3O4-L-Cys-Dox NPs as efficient drug delivery vehicles in melanoma therapy.

4.
Sci Rep ; 12(1): 21636, 2022 12 14.
Artículo en Inglés | MEDLINE | ID: mdl-36517531

RESUMEN

Microscopic evaluation of tissue sections stained with hematoxylin and eosin is the current gold standard for diagnosing thyroid pathology. Digital pathology is gaining momentum providing the pathologist with additional cues to traditional routes when placing a diagnosis, therefore it is extremely important to develop new image analysis methods that can extract image features with diagnostic potential. In this work, we use histogram and texture analysis to extract features from microscopic images acquired on thin thyroid nodule capsules sections and demonstrate how they enable the differential diagnosis of thyroid nodules. Targeted thyroid nodules are benign (i.e., follicular adenoma) and malignant (i.e., papillary thyroid carcinoma and its sub-type arising within a follicular adenoma). Our results show that the considered image features can enable the quantitative characterization of the collagen capsule surrounding thyroid nodules and provide an accurate classification of the latter's type using random forest.


Asunto(s)
Adenoma , Neoplasias de la Tiroides , Nódulo Tiroideo , Humanos , Nódulo Tiroideo/diagnóstico por imagen , Nódulo Tiroideo/patología , Diagnóstico Diferencial , Bosques Aleatorios , Cápsulas , Neoplasias de la Tiroides/diagnóstico por imagen , Neoplasias de la Tiroides/patología , Adenoma/patología
5.
Proc Natl Acad Sci U S A ; 119(47): e2214662119, 2022 11 22.
Artículo en Inglés | MEDLINE | ID: mdl-36375085

RESUMEN

Second harmonic generation microscopy (SHG) is generally acknowledged as a powerful tool for the label-free three-dimensional visualization of tissues and advanced materials, with one of its most popular applications being collagen imaging. Despite the great need, progress in super-resolved SHG imaging lags behind the developments reported over the past years in fluorescence-based optical nanoscopy. In this work, we demonstrate super-resolved re-scan SHG, qualitatively and quantitatively showing on collagenous tissues the available resolution advantage over the diffraction limit. We introduce as well super-resolved re-scan two-photon excited fluorescence microscopy, an imaging modality not explored to date.


Asunto(s)
Microscopía de Generación del Segundo Armónico , Microscopía de Generación del Segundo Armónico/métodos , Microscopía Fluorescente/métodos , Colágeno , Fotones , Cintigrafía
6.
Micromachines (Basel) ; 13(10)2022 Sep 22.
Artículo en Inglés | MEDLINE | ID: mdl-36295927

RESUMEN

In the present study, we report the development and characterization of composite layers (by spin coating) based on magnesium-doped hydroxyapatite in a chitosan matrix, (Ca10-xMgx(PO4)6(OH)2; xMg = 0, 0.08 and 0.3; HApCh, 8MgHApCh and 30MgHApCh). The MgHApCh composite layers were investigated using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray photoelectron spectroscopy (XPS) techniques. The in vitro biological evaluation included the assessment of their cytotoxicity on MG63 osteoblast-like cells and antifungal activity against Candida albicans ATCC 10231 fungal cell lines. The results of the physico-chemical characterization highlighted the obtaining of uniform and homogeneous composite layers. In addition, the biological assays demonstrated that the increase in the magnesium concentration in the samples enhanced the antifungal effect but also decreased their cytocompatibility. However, for certain optimal magnesium ion concentrations, the composite layers presented both excellent biocompatibility and antifungal properties, suggesting their promising potential for biomedical applications in both implantology and dentistry.

7.
Sci Data ; 9(1): 376, 2022 07 02.
Artículo en Inglés | MEDLINE | ID: mdl-35780180

RESUMEN

Second harmonic generation (SHG) microscopy is acknowledged as an established imaging technique capable to provide information on the collagen architecture in tissues that is highly valuable for the diagnostics of various pathologies. The polarization-resolved extension of SHG (PSHG) microscopy, together with associated image processing methods, retrieves extensive image sets under different input polarization settings, which are not fully exploited in clinical settings. To facilitate this, we introduce PSHG-TISS, a collection of PSHG images, accompanied by additional computationally generated images which can be used to complement the subjective qualitative analysis of SHG images. These latter have been calculated using the single-axis molecule model for collagen and provide 2D representations of different specific PSHG parameters known to account for the collagen structure and distribution. PSHG-TISS can aid refining existing PSHG image analysis methods, while also supporting the development of novel image processing and analysis methods capable to extract meaningful quantitative data from the raw PSHG image sets. PSHG-TISS can facilitate the breadth and widespread of PSHG applications in tissue analysis and diagnostics.


Asunto(s)
Colágeno , Microscopía de Generación del Segundo Armónico , Fijación del Tejido , Animales , Humanos , Procesamiento de Imagen Asistido por Computador
8.
ACS Omega ; 7(13): 11353-11362, 2022 Apr 05.
Artículo en Inglés | MEDLINE | ID: mdl-35415325

RESUMEN

Scattering-type scanning near-field optical microscopy (s-SNOM) has emerged over the past years as a powerful characterization tool that can probe important properties of advanced materials and biological samples in a label-free manner, with spatial resolutions lying in the nanoscale realm. In this work, we explore such usefulness in relationship with an interesting class of materials: polymer-coated gold nanoparticles (NPs). As thoroughly discussed in recent works, the interplay between the Au core and the polymeric shell has been found to be important in many applications devoted to biomedicine. We investigate bare Au NPs next to polystyrenesulfonate (PSS) and poly(diallyldimethylammonium chloride) (PDDA) coated ones under 532 nm laser excitation, an wavelength matching the surface plasmon band of the custom-synthesized nanoparticles. We observe consistent s-SNOM phase signals in the case of bare and shallow-coated Au NPs, whereas for thicker shell instances, these signals fade. For all investigated samples, the s-SNOM amplitude signals were found to be very weak, which may be related to reduced scattering efficiency due to absorption of the incident beam. We consider these observations important, as they may facilitate studies and applications in nanomedicine and nanotechnology where the precise positioning of polymer-coated Au NPs with nanoscale resolution is needed besides their dielectric function and related intrinsic optical properties, which are also quantitatively available with s-SNOM.

9.
Front Med (Lausanne) ; 9: 839786, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35280872

RESUMEN

Two-photon microscopy techniques are non-linear optical imaging methods which are gaining momentum in the investigation of fixed tissue sections, fresh tissue or even for in vivo experiments. Two-photon excited fluorescence and second harmonic generation are two non-linear optical contrast mechanisms which can be simultaneously used for offering complementary information on the tissue architecture. While the former can originate from endogenous autofluorescence sources (e.g., NADH, FAD, elastin, keratin, lipofuscins, or melanin), or exogenous eosin, the latter is generated in fibrillar structures within living organisms (e.g., collagen and myosin). Here we test the ability of both these contrast mechanisms to highlight features of the extramammary Paget disease on fixed tissue sections prepared for standard histological examination using immunohistochemical markers and hematoxylin and eosin staining. We also demonstrate the label-free abilities of both imaging techniques to highlight histological features on unstained fixed tissue sections. The study demonstrated that two-photon microscopy can detect specific cellular features of the extramammary Paget disease in good correlation with histopathological results.

10.
Materials (Basel) ; 16(1)2022 Dec 27.
Artículo en Inglés | MEDLINE | ID: mdl-36614609

RESUMEN

Spectroscopic characteristics of RE3+ ions (RE = Sm, Dy, and Pr) doped in partially disordered Ca3Nb1.6875Ga3.1875O12-CNGG and Ca3Li0.275Nb1.775Ga2.95O12-CLNGG crystals are reviewed in detail to assess their prospects as laser crystals with emission in the visible spectral domain. All investigated crystals were grown using the Czochralski crystal growth technique. High-resolution absorption and emission measurements at different temperatures, as well as emission dynamics measurements, were performed on the grown crystals. The spectroscopic and laser emission characteristics of the obtained crystals were determined based on the Judd-Ofelt parameters. The obtained results indicate that CNGG:RE3+ and CLNGG:RE3+ (RE = Sm, Dy, and Pr) crystals can be promising materials for lasers in the visible range.

11.
Biomed Opt Express ; 12(9): 5829-5843, 2021 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-34692218

RESUMEN

Second harmonic generation (SHG) microscopy has emerged over the past two decades as a powerful tool for tissue characterization and diagnostics. Its main applications in medicine are related to mapping the collagen architecture of in-vivo, ex-vivo and fixed tissues based on endogenous contrast. In this work we present how H&E staining of excised and fixed tissues influences the extraction and use of image parameters specific to polarization-resolved SHG (PSHG) microscopy, which are known to provide quantitative information on the collagen structure and organization. We employ a theoretical collagen model for fitting the experimental PSHG datasets to obtain the second order susceptibility tensor elements ratios and the fitting efficiency. Furthermore, the second harmonic intensity acquired under circular polarization is investigated. The evolution of these parameters in both forward- and backward-collected SHG are computed for both H&E-stained and unstained tissue sections. Consistent modifications are observed between the two cases in terms of the fitting efficiency and the second harmonic intensity. This suggests that similar quantitative analysis workflows applied to PSHG images collected on stained and unstained tissues could yield different results, and hence affect the diagnostic accuracy.

12.
Gigascience ; 9(11)2020 11 24.
Artículo en Inglés | MEDLINE | ID: mdl-33231675

RESUMEN

BACKGROUND: In recent years, a variety of imaging techniques operating at nanoscale resolution have been reported. These techniques have the potential to enrich our understanding of bacterial species relevant to human health, such as antibiotic-resistant pathogens. However, owing to the novelty of these techniques, their use is still confined to addressing very particular applications, and their availability is limited owing to associated costs and required expertise. Among these, scattering-type scanning near field optical microscopy (s-SNOM) has been demonstrated as a powerful tool for exploring important optical properties at nanoscale resolution, depending only on the size of a sharp tip. Despite its huge potential to resolve aspects that cannot be tackled otherwise, the penetration of s-SNOM into the life sciences is still proceeding at a slow pace for the aforementioned reasons. RESULTS: In this work we introduce SSNOMBACTER, a set of s-SNOM images collected on 15 bacterial species. These come accompanied by registered Atomic Force Microscopy images, which are useful for placing nanoscale optical information in a relevant topographic context. CONCLUSIONS: The proposed dataset aims to augment the popularity of s-SNOM and for accelerating its penetration in life sciences. Furthermore, we consider this dataset to be useful for the development and benchmarking of image analysis tools dedicated to s-SNOM imaging, which are scarce, despite the high need. In this latter context we discuss a series of image processing and analysis applications where SSNOMBACTER could be of help.


Asunto(s)
Procesamiento de Imagen Asistido por Computador , Humanos , Microscopía de Fuerza Atómica
13.
J Biophotonics ; 13(12): e202000262, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-32888377

RESUMEN

Polarization-resolved second harmonic generation microscopy is used to provide pixel-level angular distribution of collagen in thyroid nodule capsules. The pixel-level angular distribution is combined with textural analysis to quantify the collagen distribution in follicular adenoma (benign) and papillary thyroid carcinoma (malignant). Three second order nonlinear susceptibility tensor elements ratios, the collagen angular distribution and two parameters accounting for the collagen angular dispersion in different sized areas are extracted and corresponding images are computed in a pixel-by-pixel fashion. Subsequently, we show that texture analysis can be performed on these images to detect significant differences between the considered benign and malignant nodule capsules.


Asunto(s)
Microscopía de Generación del Segundo Armónico , Nódulo Tiroideo , Colágeno , Humanos , Nódulo Tiroideo/diagnóstico por imagen
14.
Appl Opt ; 59(23): 6925-6931, 2020 Aug 10.
Artículo en Inglés | MEDLINE | ID: mdl-32788782

RESUMEN

Papillary carcinoma is the most prevalent type of thyroid cancer. Its diagnosis requires accurate and subjective analyses from expert pathologists. Here we propose a method based on the Hough transform (HT) to detect and objectively quantify local structural differences in collagen thyroid nodule capsules. Second harmonic generation (SHG) microscopy images were acquired on non-stained histological sections of capsule fragments surrounding the healthy thyroid gland and benign and tumoral/malignant nodules. The HT was applied to each SHG image to extract numerical information on the organization of the collagen architecture in the tissues under analysis. Results show that control thyroid capsule samples present a non-organized structure composed of wavy collagen distribution with local orientations. On the opposite, in capsules surrounding malignant nodules, a remodeling of the collagen network takes place and local undulations disappear, resulting in an aligned pattern with a global preferential orientation. The HT procedure was able to quantitatively differentiate thyroid capsules from capsules surrounding papillary thyroid carcinoma (PTC) nodules. Moreover, the algorithm also reveals that the collagen arrangement of the capsules surrounding benign nodules significantly differs from both the thyroid control and PTC nodule capsules. Combining SHG imaging with the HT results thus in an automatic and objective tool to discriminate between the pathological modifications that affect the capsules of thyroid nodules across the progressions of PTC, with potential to be used in clinical settings to complement current state-of-the-art diagnostic methods.


Asunto(s)
Colágeno/química , Microscopía de Generación del Segundo Armónico/métodos , Cáncer Papilar Tiroideo/química , Glándula Tiroides/química , Neoplasias de la Tiroides/química , Nódulo Tiroideo/química , Adenocarcinoma Folicular/química , Algoritmos , Colágeno/ultraestructura , Humanos , Conformación Proteica , Estructura Secundaria de Proteína
15.
J Biophotonics ; 13(9): e202000097, 2020 09.
Artículo en Inglés | MEDLINE | ID: mdl-32483852

RESUMEN

Super-resolution microscopy techniques can provide answers to still pending questions on prokaryotic organisms but are yet to be used at their full potential for this purpose. To address this, we evaluate the ability of the rhodamine-like KK114 dye to label various types of bacteria, to enable imaging of fine structural details with stimulated emission depletion microscopy (STED). We assessed fluorescent labeling with KK114 for eleven Gram-positive and Gram-negative bacterial species and observed that this contrast agent binds to their cell membranes. Significant differences in the labeling outputs were noticed across the tested bacterial species, but importantly, KK114-staining allowed the observation of subtle nanometric cell details in some cases. For example, a helix pattern resembling a cytoskeleton arrangement was detected in Bacillus subtilis. Furthermore, we found that KK114 easily penetrates the membrane of bacterial microorganism that lost their viability, which can be useful to discriminate between living and dead cells.


Asunto(s)
Bacterias , Colorantes Fluorescentes , Microscopía Fluorescente , Rodaminas , Coloración y Etiquetado
16.
Materials (Basel) ; 12(18)2019 Sep 17.
Artículo en Inglés | MEDLINE | ID: mdl-31533217

RESUMEN

Despite intense research on high entropy films, the mechanism of film growth and the influence of key factors remain incompletely understood. In this study, high entropy films consisting of five elements (FeCoNiCrAl) with columnar and nanometer-scale grains were prepared by magnetron sputtering. The high entropy film growth mechanism, including the formation of the amorphous domain, equiaxial nanocrystalline structure and columnar crystal was clarified by analyzing the microstructure in detail. Besides, the impacts of the important deposition parameters including the substrate temperature, the powder loaded in the target, and the crystal orientation of the substrate on the grain size and morphology, phase structure, crystallinity and elemental uniformity were revealed. The mechanical properties of high entropy films with various microstructure features were investigated by nanoindentation. With the optimized grain size and microstructure, the film deposited at 350 °C using a power of 100 W exhibits the highest hardness of 11.09 GPa. Our findings not only help understanding the mechanisms during the high entropy film deposition, but also provide guidance in manufacturing other novel high entropy films.

17.
Materials (Basel) ; 12(16)2019 Aug 18.
Artículo en Inglés | MEDLINE | ID: mdl-31426589

RESUMEN

This paper provides a new method to compare and then reveal the vacancy sink efficiencies quantitively between different hetero-interfaces with a shared Cu layer in one sample, in contrast to previous studies, which have compared the vacancy sink efficiencies of interfaces in different samples. Cu-Nb-Cu-V nanoscale metallic multilayer composites (NMMCs) containing Cu/V and Cu/Nb interfaces periodically were prepared as research samples and bombarded with helium ions to create vacancies which were filled by helium bubbles. A special Cu layer shared by adjoining Cu/V and Cu/Nb interfaces exists, in which the implanted helium concentration reaches its maximum and remains nearly constant with a well-designed incident energy. The results show that bubble-denuded zones (BDZ) close to interfaces exist, and that the width of the BDZ close to the Cu/V interface is less than that of Cu/Nb interface. This result is explained by one-dimensional diffusion theory, and the ratio of vacancy sink efficiency between Cu/V and Cu/Nb interfaces is calculated. Conclusively, Cu/Nb interfaces are more efficient than Cu/V interfaces in eliminating vacancies induced by radiation.

18.
ACS Appl Mater Interfaces ; 11(23): 20987-20994, 2019 Jun 12.
Artículo en Inglés | MEDLINE | ID: mdl-31117446

RESUMEN

Nonlinear optical (NLO) crystals with incongruent melting of La xY ySc z(BO3)4 ( x + y + z = 4) (LYSB)-type were grown for the first time, to the best of our knowledge, by the Czochralski method. A special thermal assembly was used and the melt composition, growth direction, and the pulling and rotation rates have been optimized. Good optical quality LYSB crystal with a diameter of about 13 mm and a length of 25 mm has been grown from the La0.765Y0.485Sc2.75(BO3)4 starting melt composition, along the c-axis direction, using a slow rotation rate of 8-10 rpm and a high pulling rate of 2 mm/h. The grown crystal has an acentric huntite-type structure (space group R32, Z = 3) with cell dimensions a = 9.8098(4) Å and c = 7.9802(3) Å, and its chemical composition was determined to be La0.78Y0.32Sc2.90(BO3)4. The optical transmission and the refractive indices were determined, and the second harmonic generation (SHG) and sum frequency generation (SFG, ω + 2ω) properties were reported. The laser damage threshold was also determined to be ∼2.0 GW/cm2 at 1064 nm (6 ns pulses). The main nonlinear properties of Czochralski-grown LYSB crystal were found to be similar to those of flux-grown LYSB, and comparable to YAl3(BO3)4 (YAB) nonlinear properties. The big advantage of Czochralski-grown LYSB crystals is that they can be grown with large size and high quality, making them promising candidates for various NLO applications, including frequency conversion of high-average power radiation sources.

19.
Biomed Opt Express ; 9(8): 3923-3936, 2018 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-30338165

RESUMEN

Quantitative second harmonic generation microscopy was used to investigate collagen organization in the fibrillar capsules of human benign and malignant thyroid nodules. We demonstrate that the combination of texture analysis and second harmonic generation images of collagen can be used to differentiate between capsules surrounding the thyroid follicular adenoma and papillary carcinoma nodules. Our findings indicate that second harmonic generation microscopy can provide quantitative information about the collagenous capsule surrounding both the thyroid and thyroid nodules, which may complement traditional histopathological examination.

20.
Nanomedicine ; 14(1): 47-50, 2018 01.
Artículo en Inglés | MEDLINE | ID: mdl-28887212

RESUMEN

We present a novel method for nanoscale reconstruction of complex refractive index by using scattering-type Scanning Near-field Optical Microscopy (s-SNOM). Our method relies on correlating s-SNOM experimental image data with computational data obtained through simulation of the classical oscillating point-dipole model. This results in assigning a certain dielectric function for every pixel of the s-SNOM images, which further results in nanoscale mapping of the refractive index. This method is employed on human erythrocytes to demonstrate the approach in a biologically relevant manner. The presented results advance the current knowledge on the capabilities of s-SNOM to extract quantitative information with nanoscale resolution from optical data sets with biological application.


Asunto(s)
Eritrocitos/citología , Nanotecnología/métodos , Imagen Óptica/métodos , Refractometría , Humanos , Microscopía/métodos , Dispersión de Radiación
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