Samples were fixed in 2% w/v glutaraldehyde + 0.5% w/v paraformaldehyde (v/v) in a 50 mM sodium cacodylate buffer, pH 7.2 at room temperature and under vacuum. After 4 hours, the fixative solution was refreshed and samples were kept at 4°C for 26 days. Samples were rinsed twice in 50 mM sodium cacodylate buffer (pH 7.2) and postfixed in 2% w/v osmium tetroxide in water for 1.5 hours at room temperature. Samples were rinsed 3 times in demineralized water and dehydrated using a graded water/ethanol series (10, 20, 30, 40, 50, 60, 70, 80, 90, 96% (v/v)). Samples were first incubated in propylene oxide (Electron Microscopy Sciences, Hatfield PA, USA) twice for 1 hour, then in a propylene oxide /Epon series over several days at 4°C, positioned in their silicone embedding molds and polymerized for 48 hours at 60°C. Thin sections were cut using a Reichert Ultracut E (Leica Microsystems) and contrasted using Uranyless and lead citrate (Delta Microscopies, France). Samples were observed using a Hitachi HT7700 instrument.
Ht7700 instrument
Manufactured by Hitachi
Sourced in Japan
The HT7700 is a high-resolution transmission electron microscope (TEM) designed and manufactured by Hitachi. The core function of the HT7700 is to provide advanced imaging and analysis capabilities for materials science research and development.
Automatically generated - may contain errors
Lab products found in correlation
Propylene oxide, by Electron Microscopy Sciences (1 mentions)
Reichert ultracut e, by Leica Microsystems (1 mentions)
S 3400n microscope, by Hitachi (1 mentions)
Nanobrook 90plus zeta, by Brookhaven Instruments (1 mentions)
Dpx 400 mhz spectrometer, by Bruker (1 mentions)
Glucose oxidase gox, by Merck Group (1 mentions)
Rf 5301pc fluorescence spectrometer, by Shimadzu (1 mentions)
Delsatm nano system, by Beckman Coulter (1 mentions)
500 mhz spectrometer, by Bruker (1 mentions)
Infinite m1000 pro, by Tecan (1 mentions)
5 protocols using ht7700 instrument
1
Comprehensive Electron Microscopy Sample Preparation
2
Ultrastructural Analysis of Spinal Cord
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All electron microscopy specimens were fixed with 2.5 % glutaraldehyde. An NT3 antibody (Proteintech 18084-1-AP, 1:100) was used for immunoelectron microscopy (IEM) analysis. The sampling location of transmission electron microscope (TEM) was the central region of the SCI area. The images of scanning electron microscope (SEM) and IEM were performed using a Hitachi S-3400 N microscope (Tokyo, Japan). The TEM images were acquired using a Hitachi HT7700 instrument (Tokyo, Japan).
3
Synthesis and Characterization of Functional Materials
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All reagents were purchased commercially and used without further purification unless otherwise noted. All reactions were performed in a normal air atmosphere unless otherwise stated. Glucose oxidase (GOx, 100 unit/mg protein) was purchased from Sigma-Aldrich. Column chromatography was performed with silica gel (200–300 mesh) produced by Qingdao Marine Chemical Factory, Qingdao (China). NMR spectra were recorded on a Bruker DPX 400 MHz spectrometer using the internal standard tetramethylsilane (TMS). HRMS data were recorded on a JMS-SX102A (FAB) or via LC/MSD TOF. Dynamic light scattering (DLS) measurements were taken using a NanoBrook 90Plus Zeta (Brookhaven Instruments Corporation, New York, NY, USA) with a 200 mW polarized laser source (λ = 514 nm). Zeta potential measurements were recorded at 25 °C using a NanoBrook 90Plus Zeta (Brookhaven Instruments Corporation, New York, NY, USA). Transmission electron microscope (TEM) imaging was performed using a HITACHI HT-7700 instrument. The UV–Vis absorption spectra were recorded on a spectrometer (UV1800PC, Jinghua, Shanghai, China). The EPR (electron paramagnetic resonance) spectra were recorded on JON Bruker BioSpin GmbH. The excitation and emission spectra were measured on a SHIMADZU RF-5301PC fluorescence spectrometer.
4
Characterization of Optimized SSM-SNEDDS Droplets
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The droplets of the optimized SSM–SNEDDS formulation were examined through TEM. Specifically, SSM–SNEDDS was diluted 400 times with Mini-Q water (0.5%, w/v) and dropped on a 300-mesh copper grid coated with carbon. After negative staining with 2% (w/v) phosphotungstic acid (PTA) solution, excess PTA was removed using Mini-Q water. The copper grid was then placed in a vacuum drying oven for 2 days. The dried grid was observed through TEM using an HT7700 instrument (Hitachi, Tokyo, Japan) at a voltage of 100 kV. The ImageJ (National Institutes of Health, Bethesda, MD, USA) was used to confirm the droplet size.
5
Characterization of Nanoparticle Samples
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NMR spectra were recorded on a Bruker 500 MHz Spectrometer (Bruker Corporation, Karlsruhe, Germany), with working frequencies of 500 MHz for 1H and 125 MHz for 13C. The residual signals from DMSO-d6 (1H: δ 2.50 ppm; 13C: δ 39.52 ppm) or CDCl3 (1H: δ 7.26 ppm; 13C: δ 77.16 ppm) were used as internal standards. Negative-stained transmission electron microscope (TEM) images were taken on an HT7700 instrument (Hitachi Ltd., Tokyo, Japan, 80 kV). The samples for negative-stained TEM were prepared by dropping a droplet of the sample solution onto a TEM grid (copper grid, 300 mesh, coated with carbon film). The ζ potentials and dynamic light scattering (DLS) measurements of the nanoparticles at different fabrication stages were measured using a DelsaTM Nano system (Beckman Coulter, Brea, CA, USA). Cell culture was carried out in an incubator with a humidified atmosphere of 5% CO2 at 37°C. UV-Vis spectra were recorded with a Shimadzu 1750 UV-visible spectrophotometer (Kyoto, Japan) at 298 K. The DOX absorbance and the MTT data were obtained from a microplate reader (Tecan Infinite M1000 Pro, Männedorf Switzerland), with a λ range from 230 nm to 1,000 nm. The surface area was measured by nitrogen physisorption (Quantachrome, Autosorb-iQ) based on the Brunauer–Emmet–Teller (BET) method (ASAP 2020, Micromeritics Inc, GA, USA).
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