The morphology of niosome was visualized by atomic force microscopy (AFM; Alpha300 RA; WITec, Ulm, Germany) and transmission electron microscopy (TEM; JEOL 2100F; Tokyo, Japan). The sizes of the NPs were measured by dynamic light scattering (DLS). Zeta potential was measured to evaluate the surface charge on the particles. Samples were diluted in double-distilled water for both DLS and zeta sizer and were analyzed in Malvern Zetasizer ZS 90 (Malvern Instruments, Malvern, UK). For TEM analysis, the samples were diluted in double-distilled water, loaded on copper grid, and kept under vacuum for drying. After drying, the samples were analyzed under JEOL 2100F. For the AFM analysis, the samples were diluted in double-distilled water, and 5 µL of the sample was loaded onto a mica sheet and dried. The samples were analyzed in WITEC ALPHA 300RA. The resonant frequency of the tip was 80 kHz with a spring constant of 40 N/m, and imaging was performed in air by tapping mode. The analysis of topography data was performed by Project 4 software.
Alpha300ra
Manufactured by WITec
Sourced in Germany
The Alpha300RA is a research-grade confocal Raman microscope system designed for advanced materials analysis. It features high-resolution imaging, precise spectroscopic measurements, and a flexible modular design to accommodate a variety of sample types and research applications.
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Lab products found in correlation
Uhts 300, by WITec (11 mentions)
Control four, by WITec (5 mentions)
Opus 7, by Bruker (4 mentions)
Du401a bv, by WITec (3 mentions)
Oil immersion objective, by Zeiss (3 mentions)
Project five plus, by WITec (3 mentions)
Project four 4, by WITec (3 mentions)
D8 advance, by Bruker (2 mentions)
Vertex 70, by Bruker (2 mentions)
Project four, by WITec (2 mentions)
K alpha, by Thermo Fisher Scientific (2 mentions)
Sigma 300, by Zeiss (2 mentions)
Igor pro 7, by Wavemetrics (1 mentions)
Projectfive 5.1 plus, by WITec (1 mentions)
Escalab xi, by Thermo Fisher Scientific (1 mentions)
4200a scs, by Tektronix (1 mentions)
S 4800, by Hitachi (1 mentions)
Fluorescence spectrometer, by Shimadzu (1 mentions)
Model jem 2100, by JEOL (1 mentions)
Nicolet 370 spectrophotometer, by Thermo Fisher Scientific (1 mentions)
69 protocols using alpha300ra
1
Niosome Morphology Analysis by Advanced Microscopy
2
Confocal Raman Mapping of Calcifications
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A confocal Raman microscope (alpha300RA, WITec, Ulm, Germany) with a frequency-doubled Nd:YAG laser (532 nm) excitation source was used (maximum power: 75 mW). The charge-coupled device detector (DU401A-BV, Andor, UK) was placed behind the spectrometer (UHTS 300, WITec, Ulm, Germany) with a grating of 600 g/mm (Blaze wavelength = 500 nm). Samples were placed on a multi-axis piezo scanner and a motorized large-area stage for sample positioning and imaged using a water immersion 60× objective [Nikon, numerical aperture (NA) = 1.0]. The lateral resolution was 0.61 λ/NA = ~325 nm. The typical mapping scanning step size used was 1 μm (though ranged from 0.5 to 2 μm) with a typical integration time of 0.23 s per step (though going as high as 4 s per step in rare cases).
Mapping data from 117 regions (from 96 individual calcifications and 5 particle-containing regions) containing Raman-detected calcifications with pathological continuity were analyzed. All Raman data were analyzed using WITec Project Plus version 4.1 and, where indicated, Igor Pro 7 (WaveMetrics Inc., Lake Oswego, OR, USA). Raman data processing is elaborated in Supplementary Text.
Mapping data from 117 regions (from 96 individual calcifications and 5 particle-containing regions) containing Raman-detected calcifications with pathological continuity were analyzed. All Raman data were analyzed using WITec Project Plus version 4.1 and, where indicated, Igor Pro 7 (WaveMetrics Inc., Lake Oswego, OR, USA). Raman data processing is elaborated in Supplementary Text.
3
Confocal Raman Microscopy of Pharmaceutical Samples
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Confocal Raman microscopy was performed using WITec Alpha 300-RA confocal Raman microscopy equipment (WITec, Ulm, Germany). The excitation laser wavelength corresponded to λ = 532 nm and the nominal power was adjusted to 45 mW to avoid sample decomposition. Raman spectra of the pure components were obtained by averaging a set of 100 spectra with an integration time of 0.512 s for each spectrum. Two-dimensional confocal Raman microscopy images of the PM and TA-NC samples were collected at random locations of 150 × 150 µm areas with 50 × 50 point grids defining the bitmap image (2500 pixels). Individual Raman spectra were collected for each pixel in the selected areas with an integration time of 0.112 s. The spectrometer operating with 600 lines/mm grating allowed us to obtain spectra with a resolution of ~4 cm−1 in the range of 70–4000 cm−1. All images were collected at an optical resolution limit of ~300 nm. True component analysis was used to map the spatial distribution of mTA and P188 in the PM or TA-NC samples using ProjectFive 5.1 Plus software (WITec, Ulm, Germany).
4
Characterization of Hexagonal Boron Nitride
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The morphology of h-BN was characterized by using a scanning electron microscope (SEM, Hitachi S-4800). The monolayer thickness and vertical conduction of h-BN films were measured by a CAFM system (Bruker Nano GmbH, Bruker NW4). For the antioxidant test, the bare Cu foil and Cu foil covered with fully coalesced h-BN monolayer were placed on a heating plate, then the samples were heated in air at 200 oC for 10 min. Light transmission and absorption spectra were collected by a UV-Vis-NIR spectrophotometer (Agilent Technologies Cary 50000). Raman spectra were collected by a Raman microscope (WITec alpha 300RA) with a 488 nm laser. The X-ray photoelectron spectroscopy (XPS, PHI Quantera) and the Auger electron spectroscopy (AES, PHI660 system) were conducted to confirm the composition and impurity doping in the h-BN monolayer. UPS (Thermo Fischer, ESCALAB Xi+) was employed to determine the electronic work function of h-BN films. The transmission electron microscopy (TEM) investigation was carried out using a FEI Talos F200s field-emission electron microscope. The electrical measurement of I–V curve was carried out on a probe station with a Keithley 2450 system. The capacitance-voltage measurement was conducted in a semiconductor parameter analyzer (Tektronix, 4200A-SCS).
5
Multimodal Characterization of Nanomaterials
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Raman measurements were recorded at a wavelength of 514.5 nm using confocal Raman microscope by Renishaw (In Via Raman). TEM analyses of the samples were done using a JEOL JEM-2100 model. The fluorescent study was carried out with an RF-5301 PC, Shimadzu fluorescence spectrometer. Functional groups were qualitatively identified by a Fourier transform infrared spectrometer (Thermo Nicolet 370 spectrophotometer). Atomic force microscope (Witec Alpha 300RA) was used for the height profile analysis. The UV/Vis spectrometer (Ocean optics JAZ series) was used for acquiring absorption spectra. The CHNS analysis was carried out using Elementar Vario EL111 elemental analyzer.
6
Fabrication of Molecular Gate Films
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pNaSS ‘molecular-gate’ films were spin-coated from 2 to 4 wt% solutions in deionised water with Triton X-100 surfactant added at ~1 wt% relative to the amount of solvent. pNaSS solutions were filtered (polyethersulfone (PES), 0.45 μm pore size) prior to use. For all functional small-molecular donor layers, spin-coating of the respective solutions was carried out by dynamic deposition—i.e., onto the substrate at the target spin speed. Laser patterning was carried out using a WITec Alpha 300RA instrument using its stepper-motor-driven stage for scanning the sample in the laser focal plane. Continuous-wave (CW) laser excitation was used throughout.
7
Confocal Raman Mapping of PVA Hydrogels
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Each sample was hydrated for more than 2 hours and then pressed between a glass slide and a coverslip to ensure a flat surface. The coverslip was then sealed at the edges with nail polish to prevent the hydrogel from drying. A confocal Raman microscope (alpha300 RA, WITec, Germany) with 20× objective (Zeiss, Germany) was used. An Nd:YAG laser (532 nm) was used as the excitation source with the maximum power of 75 mW. Data were collected using a charge-coupled device detector (DU401A-BV, Andor, UK) behind a grating spectrometer (600 g/mm; UHTS 300, WITec, Germany). A 20-μm-resolution Raman map of 4 × 3–mm scan area was acquired with an accumulation time of 1 s per point. Each point was prebleached for 400 ms to decrease the effect of fluorescence. Cosmic ray removal and background subtraction were performed to clean the spectra. The intensity of O–H bond within the PVA and water was calculated by integrating the spectra in the range of 2800 to 3000 cm−1 and 3075 to 3625 cm−1, respectively. The ratio of PVA and water was then calculated and plotted as a heatmap shown in Fig. 5A .
8
Characterization of Shell Nacre Powder
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The particle size and morphology of shell nacre powder were studied by SEM. Shell nacre powder was mixed with potassium bromide at a ratio of 1:400, and the FTIR spectrum was recorded from 400 to 3750 cm−1 at a resolution of 4 cm−1 (FTIR Carry 600, Agilent technologies, Bangalore, India). The Raman spectrum of shell nacre powder was recorded using confocal Raman microscopy (alpha 300RA, Witec, Ulm, Germany) with a 532 nm laser. The XRD spectrum was measured (Bruker, D8 Advance, Germany) in the 2θ range of 10–70° with CuKά radiation in the incremental step of 5°, and peaks were identified using the JCPDS (Joint Committee on Powder Diffraction Standards) database. TGA analysis of shell nacre powder was carried out using SDT-2960, TA Instruments, heating the powder from room temperature to 1000 °C (heating rate of 10 °C/min) in a nitrogen atmosphere. Trace elements such as Cu, Fe, Mg, Mn, Zn, Cd, Pb, Hg, and Se were estimated using optical emission spectroscopy with inductively coupled plasma (OES-ICP) (PerkinElmer, Hopkinton, MA, USA).
9
Raman and IR Analysis of Microsections
10
Raman Imaging and Scanning Force Microscopy Analysis
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A combined Raman Imaging/Scanning Force Microscope System (WITec alpha 300 RA+, Germany) with WiTec Control FIVE 5.3 software was used for RAMAN measurements. Laser is equipped with a UHTS 300 spectrometer combined with a back-illuminated Andor Newton 970 electron multiplying charge-coupled device camera [resolution: ca. 300 to 400 nm (lateral) and 900 nm (z) with 100× objective].
The measurements were carried out at an excitation wavelength of λ = 532 nm and a laser power of 1 mW with 50 accumulations with an integration time of 0.5 s pixel−1. The samples were stacked on a glass slide. After adjusting the focus on the nonwovens at ×100 magnification, the Raman spectrum was recorded. A cosmic ray removal and a baseline correction were performed on all spectra. The peaks were then fitted with a Gaussian function using the built in routine of Origin 2016.
The measurements were carried out at an excitation wavelength of λ = 532 nm and a laser power of 1 mW with 50 accumulations with an integration time of 0.5 s pixel−1. The samples were stacked on a glass slide. After adjusting the focus on the nonwovens at ×100 magnification, the Raman spectrum was recorded. A cosmic ray removal and a baseline correction were performed on all spectra. The peaks were then fitted with a Gaussian function using the built in routine of Origin 2016.
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