Mice myocardium was fixed in electron microscope fixative fluid (Wuhan Servicebio Biotechnology Co. Ltd) at 24 h after CLP or sham operation, then staying overnight under 4°C. Then after sequential steps of washing, fixation with osmic acid, washing, dehydration with acetone, resin embedding, baking, slicing by Leica EMUC7 ultramicrotome and staining with uranyl acetate and lead citrate. Images were taken under 1400plus JEOL TEM.
1400plus tem
Manufactured by JEOL
Sourced in Germany
The 1400Plus TEM is a transmission electron microscope (TEM) manufactured by JEOL. It is designed to provide high-resolution imaging and analysis of samples at the nanoscale level. The core function of the 1400Plus TEM is to generate and transmit an electron beam through a thin specimen, creating a magnified image that can be observed and analyzed.
Automatically generated - may contain errors
Lab products found in correlation
Cm 10 tem, by JEOL (2 mentions)
Lsm 510, by Zeiss (2 mentions)
Lsm 880, by Zeiss (2 mentions)
Em uc7 ultramicrotome, by Leica (1 mentions)
Multimode 5, by Bruker (1 mentions)
Zetasizer nano s, by Malvern Panalytical (1 mentions)
Avance 3, by Bruker (1 mentions)
Anti iba1 antibody, by Fujifilm (1 mentions)
Aclar film, by Electron Microscopy Sciences (1 mentions)
Ultracut uc6, by Leica (1 mentions)
Chloroauric acid trihydrate, by Thermo Fisher Scientific (1 mentions)
Uv 1601 uv vis spectrometer, by Shimadzu (1 mentions)
8453 uv visdiode array spectrophotometer, by Agilent Technologies (1 mentions)
11 protocols using 1400plus tem
1
Ultrastructural Analysis of Sepsis Mouse Myocardium
2
Characterization of Dendritic Nanostructures
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DLS was done on a Malvern Zetasizer
Nano S equipped with a laser operating at 633 nm. The sample grids
used for electron microscopy were purchased from Electron Microscopy
Sciences (EMS, Hatfield, PA, USA). The grids were treated with a plasma
cleaner setup (20 s at 10–1 Torr) to make them hydrophilic.
For TEM measurements, samples were prepared on hydrophilic 400 mesh
carbon-coated copper grids, while 200 mesh carbon-coated copper grids
were used for the TEM tilt series. The TEM tilt series were acquired
with SerialEM, and IMOD was used for 3D reconstruction of the dendroids
from the tilt series. For cryo-TEM measurements, 400 mesh holey carbon
grids were used. For cryo-TEM, after blotting, the samples were plunged
into liquid ethane at about liquid nitrogen temperature by using a
Vitrobot system (FEI Company). Samples for (cryo-)TEM were imaged
with a 1400Plus JEOL TEM operating at 120 kV. The contrast of (cryo-)TEM
pictures was adjusted by using ImageJ-win64. Atomic force microscopy
images were recorded on a Bruker Multimode 5 using contact mode. NMR
spectra in D2O were obtained on a Bruker Avance III spectrometer
operating at 500 MHz for 1H, equipped with a 5 mm TXI probe.
Fluorescence emission spectra were acquired on a Cary Eclipse spectrophotometer.
Nano S equipped with a laser operating at 633 nm. The sample grids
used for electron microscopy were purchased from Electron Microscopy
Sciences (EMS, Hatfield, PA, USA). The grids were treated with a plasma
cleaner setup (20 s at 10–1 Torr) to make them hydrophilic.
For TEM measurements, samples were prepared on hydrophilic 400 mesh
carbon-coated copper grids, while 200 mesh carbon-coated copper grids
were used for the TEM tilt series. The TEM tilt series were acquired
with SerialEM, and IMOD was used for 3D reconstruction of the dendroids
from the tilt series. For cryo-TEM measurements, 400 mesh holey carbon
grids were used. For cryo-TEM, after blotting, the samples were plunged
into liquid ethane at about liquid nitrogen temperature by using a
Vitrobot system (FEI Company). Samples for (cryo-)TEM were imaged
with a 1400Plus JEOL TEM operating at 120 kV. The contrast of (cryo-)TEM
pictures was adjusted by using ImageJ-win64. Atomic force microscopy
images were recorded on a Bruker Multimode 5 using contact mode. NMR
spectra in D2O were obtained on a Bruker Avance III spectrometer
operating at 500 MHz for 1H, equipped with a 5 mm TXI probe.
Fluorescence emission spectra were acquired on a Cary Eclipse spectrophotometer.
3
Ultrastructural analysis of IBA1 in brain
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Postmortem brain tissue fixed in 4% PFA underwent two different routes for further processing. For ultrastructural analysis, tissue was post-fixed in Itho-buffer containing 3% glutaraldehyde (GA). Embedding in resin and sectioning was performed as described before [1 (link)]. For immuno-gold labeling, tissue samples were rehydrated, frozen and cryo-sectioned to 50 µm. These 50 µm sections were then incubated with an anti-IBA1 antibody (Wako, Online Resource Table 4) and a gold-labeled secondary antibody. After washing the section to remove unbound secondary antibodies, silver enhancement was performed to increase signal intensity in transmission electron microscopic (TEM) analysis. Sections were then fixed in Itho-buffer (with 3% GA), embedded and sectioned as described above. All ultrathin sections were transferred into a 1400Plus TEM (JEOL) operating at 120 kV.
4
Negative Staining of Liposomes for TEM
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Negative staining room temperature transmission electron microscopy (TEM) was performed as previously described [64 (link)]. Briefly, 5 µL of liposome suspension was added to previously negative glow discharged holy carbon grids (Science Service, Munich, Germany) and incubated for 10 s. After removing the excess sample with filter paper, liposomes were stained twice with 5 µL of a half-saturated uranyl acetate solution and air-dried. Samples were then transferred into a JEOL 1400Plus TEM, and images were obtained at 100 kV acceleration voltage on a TVIPS F416 4 kx4 k camera (Tietz Video and Image Processing Systems, Munich, Germany).
5
Histological and Ultrastructural Analysis of Lysosomal Disorders
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For histological analysis, 3μm thick sections were stained with basic fuchsine to evaluate the presence of intracellular glycolipid as dark blue material in contrast with reddish cytoplasm and nucleus. As indicated by the present guidelines, “disorders of the lysosomal type require electron microscopy for morphological diagnosis on tissue biopsy” (15 (link)). For this reason, samples were fixed in 2% glutaraldehyde in 0.1 M phosphate buffer, at pH 7.3, post-fixed in osmium tetroxide 1,33% in the same buffer and processed following a standard schedule for embedding in Epon resin. Ultrathin sections were obtained by a diamond knife and stained with uranyl acetate and lead hydroxide. A Philips CM-10 TEM or Jeol 1400-plus TEM were used for observation and photographic analysis.
6
Tungsten-Incorporated pDNA Nanoparticle Visualization
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Polyplex solutions were imaged using a negative staining method previously developed in our lab to visualize the pDNA in this work. Tungsten-incorporated pDNA was prepared for use in the nanoparticles. Briefly, 10 mL of sodium tungstate aqueous solution (0.15 mol/L) was added dropwise to 6 mL of HCl aqueous solution (0.8 mol/L) at room temperature. The precipitate was collected and washed to obtain the activated tungstic acid. The desired quantity of activated tungstic acid was added to an aqueous DNA solution with a W/P molar ratio of 3. The W-incorporated pDNA solution was purified by centrifugation and stored at −80 °C. One drop of aqueous polyplex solution (1.0 mg/mL) was added to a carbon-coated copper grid. The grid was dried under ambient conditions. A JEOL 1400Plus TEM was used in this study to obtain TEM images.
7
Electron Microscopy Tomography Protocols
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Electron microscopy sections were tilted and imaged within a JEOL 1400Plus TEM from ±60° over two perpendicular axes and images collect using SerialEM software (developed at University of Colorado, Boulder). Dual-axis tomograms were generated from the tilt series using IMOD (developed at the University of Colorado, Boulder)37 (link). Subtomographic averaging was performed using the IMOD package, PEET (Particle Estimation for Electron Tomography). Slices from the tomograms were viewed in IMOD and the averaged structures surface rendered in Chimera (UCSF).
8
Electron Microscopy Sample Preparation
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Cells grown on coverslips made of Aclar film (Electron Microscopy Sciences) were fixed in 4% paraformaldehyde and 2.5% glutaraldehyde with 0.1% tannic acid in 0.1 M sodium cacodylate buffer at room temperature for 30 min, postfixed in 1% OsO4 in sodium cacodylate buffer for 30 min on ice, dehydrated in graded series of ethanol, infiltrated with EPON812 resin (Electron Microcopy Sciences), and then embedded in the resin. Serial sections (∼90-nm thickness) were cut on a microtome (Ultracut UC6; Leica) and stained with 1% uranyl acetate as well as 1% lead citrate. Samples were examined on JEOL transmission electron microscope. Tomography data was collected using a JEOL 1400 Plus TEM operated at 120 kV. Double tilt series were recorded from –60° to 60° with 1° increments using SerialEM software (Mastronarde, 2005 (link)). Tomograms were reconstructed using IMOD programs (Kremer et al., 1996 (link)).
9
Synthesis and Characterization of AuNP-PVA Composites
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Chloroauric acid trihydrate and trisodium citrate dihydrate were obtained from Alfa Aesar and used without further purification. The PVA 3 mm filament (Ultimaker) was obtained from Makerpoint. A Shimadzu UV1601 UV–vis spectrometer was used for the UV–vis study. The samples were imaged in a JEOL 1400Plus TEM operating at 120 kV. The mean aspect ratio of the AuNPs was calculated by measuring approximately 800 particles from 24 micrographs. A TE instruments DMA Q800 instrument was used for the mechanical analysis of the plastics using the stress/strain program with a force ramp rate of 1 N/min at isothermal (25 °C) temperature. Four samples of the pure PVA and four samples of the AuNP–PVA materials were tested, where each sample was also flipped upside down and the stress/strain was recorded again. The average of the measurements was used in Figure 2c with the standard deviation shown as the error bar.
10
Ultrastructural Analysis of the C. elegans Nerve Ring
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unc-42(e270) was fixed as previously described (White et al., 1986 (link)). These fixed worms were then cut into 50 nm sections using RMC Powertome XL and collected onto grids. The nerve ring region of unc-42(e270) was then imaged either manually with a Phillips CM10 TEM or automatically with a JEOL 1400Plus TEM and the SerialEM software. Sections were then aligned and montaged, all of the axons in the nerve ring were serially traced, and synapses were annotated using the TrakEM2 software (Cardona et al., 2012 (link)). The region imaged, reconstructed, and annotated was ~15 μm in length and included 309 serial sections. Neurons were identified by characteristic synaptic and/or morphological features together with relative cell body position (Supplementary file 6 ). Synapse counts and axon adjacency counts were then extracted using scripts kindly provided by Christopher Brittin (Brittin et al., 2018 (link)). To compare to the unc-42(e270) synapse and axon adjacency counts to the previously described wild type (N2U) (Cook et al., 2019 (link)), the sections were aligned from the beginning of the RMEV neuron nucleus, a neuron that is easily identifiable based on morphology and position, to the anterior end of the nerve ring.
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