The largest database of trusted experimental protocols

Jem 1011 transmission electron microscope

Sourced in United States, Morocco

The JEM 1011 is a transmission electron microscope (TEM) designed and manufactured by JEOL. It is capable of producing high-resolution images of samples at the nanoscale level. The TEM functions by transmitting a beam of electrons through a thin specimen, allowing for the observation and analysis of the internal structure and composition of materials.

Automatically generated - may contain errors

10 protocols using jem 1011 transmission electron microscope

1

Ultrastructural Analysis of Peri-Infarct Cortex

Check if the same lab product or an alternative is used in the 5 most similar protocols
TEM was performed as desribed previously (Begum et al., 2018 (link)). Briefly, mice were transcardially perfused with PBS followed by fixation with 2.5% glutaraldehyde for 24 h. After fixation, the brain was sectioned into 1 mm thick slices and post fixed in 2.5% glutaraldehyde in PBS. Under a dissection microscope, tissue punches were taken to capture the IL and CL peri-infarct areas in the cortex. Tissues were washed three times in PBS then, post-fixed in 1% osmium tetroxide with 1% potassium ferricyanide for 1 h. Following three additional PBS washes, the pellet was dehydrated through a graded series of 30-100% ethanol, 100% propylene oxide and then infiltrated in 1:1 mixture of propylene oxide: Polybed 812 epoxy resin for 1 h. After several changes of 100% resin over 24 hrs, pellet was embedded in a final change of resin, cured at 37°C overnight, followed by additional hardening at 65°C for two more days. Ultrathin (70 nm) sections were collected on 200 mesh copper grids, stained with 2% uranyl acetate in 50% methanol for 10 minutes, followed by 1% lead citrate for 7 min. Sections were imaged using a JEOL JEM 1011 transmission electron microscope (Peabody, MA) at 80 kV fitted with a side mount AMT 2k digital camera (Advanced Microscopy Techniques, Danvers, MA).
+ Open protocol
+ Expand
2

Transmission Electron Microscopy Sample Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell monolayers were fixed for 2 hours in 2% glutaraldehyde in 0.1 mol/L sodium cacodylate buffer, pH 7.4 (Electron Microscopy Sciences, Hatfield, PA), on Transwell membrane supports and then rinsed in 0.1 mol/L sodium cacodylate buffer. Specimens were postfixed with 1% osmium tetroxide for 1 hour at room temperature, rinsed again in 0.1 mol/L sodium cacodylate buffer, dehydrated through a graded series of 100% ethanol concentrations, dehydrated briefly in 100% propylene oxide, and pre-infiltrated overnight in a 1:1 mix of eponate resin (Ted Pella, Inc, Redding, CA) and propylene oxide. The following day, specimens were infiltrated in 100% eponate resin, embedded in flat molds with 100% eponate, and allowed to polymerize overnight at 60°C. Thin (70-nm) sections were cut using a Leica (Leica Biosystems, Wetzlar, Germany) EM UC7 ultramicrotome, collected onto formvar-coated grids, stained with uranyl acetate and lead citrate, and examined on a JEOL (Peabody, MA) JEM 1011 transmission electron microscope at 80 kV. Images were collected using an Advanced Microscopy Techniques digital imaging system (Advanced Microscopy Techniques, Woburn, MA).
+ Open protocol
+ Expand
3

Immunoelectron Microscopy of Tau Aggregates

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunoelectron microscopy was performed in the Microscopy Core of the Center for Systems Biology/Program in Membrane Biology (MGH, Boston, USA). The sarkosyl-insoluble fractions were resuspended in PBS, placed on formvar-carbon coated Ni grids and allowed to adsorb for 10 minutes. They were placed on drops of tau46 antibody solution (1:25, Cell Signaling Technology) for 1 hour at room temp, then rinsed on drops of PBS and placed on drops of goat-anti-mouse 10 nm gold (Ted Pella, Redding, CA, USA) for 1 hour. They were rinsed on drops of distilled water and stained for 1 minute on drops of 2% phosphotungstic acid (Electron Microscopy Sciences, Hatfield, PA, USA). Grids were examined in a JEOL JEM 1011 transmission electron microscope at 80 kV. Images were collected using an AMT digital imaging system (Advanced Microscopy Techniques, Danvers, MA, USA).
+ Open protocol
+ Expand
4

Transmission Electron Microscopy (TEM) Tissue Fixation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissue specimens were fixed in 2% paraformaldehyde and 2.5% glutaraldehyde in 0.1 M sodium phosphate buffer (pH 7.4, Electron Microscopy Sciences, Hatfield, PA), overnight at 4 °C, then rinsed several times in 0.1 M cacodylate buffer. Specimens were post-fixed in 1.0% osmium tetroxide in cacodylate buffer for one hour at room temperature, rinsed several times in buffer, dehydrated through a graded series of ethanols to 100%, dehydrated briefly in 100% propylene oxide, and pre-infiltrated in a 1:1 mix of Eponate resin (Ted Pella, Redding, CA) and propylene oxide overnight on a gentle rotator. The following day, specimens were infiltrated with fresh Eponate resin for several hours, embedded in flat molds with fresh Eponate and allowed to polymerize overnight at 60 °C. Thin (70 nm) sections were cut using a Leica EM UC7 ultramicrotome, collected onto formvar-coated grids, stained with uranyl acetate and Reynold’s lead citrate and examined in a JEOL JEM 1011 transmission electron microscope at 80 kV. Images were collected using an AMT digital imaging system (Advanced Microscopy Techniques, Danvers, MA).
+ Open protocol
+ Expand
5

Transmission Electron Microscopy of Liver Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cultured cells or 1 mm3 cubes of the liver tissue were fixed overnight in 2.5% glutaraldehyde in PBS at 4°C, washed, and post-fixed in aqueous 1% OsO4, 1% K3Fe(CN)6. Following 3 PBS washes, the samples were dehydrated through a graded series of 30–100% ethanol, 100% propylene oxide then infiltrated in 1:1 mixture of propylene oxide:Polybed 812 epoxy resin (Polysciences, Warrington, PA) overnight at room temperature. After several changes of 100% resin over 24 h, the samples was embedded in molds, cured at 37°C overnight, followed by additional hardening at 65°C for 2 days. Ultrathin (70 nm) sections were collected on 200 mesh copper grids, stained with 2% uranyl acetate in 50% methanol (10 min), followed by 1% lead citrate (7 min). Sections were imaged using a JEOL JEM 1011 transmission electron microscope (Peabody, MA) at 80 kV fitted with a side mount AMT 2 k digital camera (Advanced Microscopy Techniques, Danvers, MA).
+ Open protocol
+ Expand
6

Transmission Electron Microscopy of Exosomes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Approximately 5 μL of exosome sample was applied to the surface of Electron Microscopy Sciences formvar-coated, carbon-stabilized 3 mm copper grids. Each sample was left on a grid for 5 minutes before the excess liquid was wicked off using the edge of a piece of filter paper. Next, 5 μL of Electron Microscopy Sciences 2% aqueous uranyl acetate stain was applied to the grid for 1 minute as a negative stain, excess solution was wicked off, and the samples were allowed to air-dry before observation. Imaging was done using a JEOL JEM-1011 transmission electron microscope and images were collected using an Advanced Microscopy Techniques XR50S-A digital camera.
+ Open protocol
+ Expand
7

Ultrastructural Analysis of CSF Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols
Following a centrifugation at 14,000 rpm for 20 minutes, pellets obtained from CSF samples were fixed in 2.0% glutaraldehyde in 0.1 M sodium cacodylate buffer, pH 7.4 (Electron Microscopy Sciences) for one hour at room temperature on a rocker. They were rinsed in cacodylate buffer, gently scraped and pelleted and post-fixed in 1.0% osmium tetroxide in cacodylate buffer for one hour on ice. They were rinsed in buffer and stabilized with a small amount of 2% agarose in PBS to hold them together. They were then dehydrated through a graded series of ethanol to 100%, followed by propylene oxide, 100%. They were infiltrated with Epon resin (Ted Pella) in a 1:1 solution of Epon:propylene oxide overnight on a rocker at room temperature. The following day they were placed in fresh Epon for several hours and then embedded in Epon overnight at 60 °C. Thin sections were cut on a Leica EM UC7 ultramicrotome, collected on formvar-coated grids, stained with uranyl acetate and lead citrate and examined in a JEOL JEM 1011 transmission electron microscope at 80 kV. Images were collected using an AMT digital imaging system (Advanced Microscopy Techniques).
+ Open protocol
+ Expand
8

Cryosectioning and Dual Immunolabeling of PC12 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
PC12 cells were washed and fixed in a solution containing 4 % paraformaldehyde and 0.2 % glutaraldehyde in 1X PBS. Following 5 washes, cells were pelleted, resuspended in warm 2 % agarose, cut into small blocks and incubated with 2.3 M sucrose at 4 °C for overnight. Ultrathin cryosections were generated on a Leica EM FCS at −80 °C and collected on the formvar-carbon coated nickel grids. For double immunolabeling, grids were first blocked on drops of 1 % BSA and 5 % goat serum and then incubated with mouse anti-Syt-1 antibody for 1 h followed by anti-mouse secondary antibody coupled with 10 nm gold particle for 1 h. After rinsing, grids were incubated again with rabbit anti-APP antibody for 1 h followed by anti-rabbit secondary antibody coupled with 15 nm gold particles. Grids were washed, stained on drops of Tylose and Uranyl acetate and then allowed to dry. The grids were examined at 80 kV in a JEOL JEM 1011 transmission electron microscope and the images were acquired using an AMT digital imaging system (Advanced Microscopy Techniques, Danvers, MA).
+ Open protocol
+ Expand
9

Astrocyte Ultrastructural Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Rat cortical astrocytes or pellets from astrocyte-conditioned media were
fixed in 2.0% glutaraldehyde in 0.1 M sodium cacodylate buffer, pH 7.4
(Electron Microscopy Sciences, Hatfield, PA) for one hour at room temperature on
a rocker. They were rinsed in cacodylate buffer, gently scraped and pelleted and
post-fixed in 1.0% osmium tetroxide in cacodylate buffer for one hour on
ice. They were rinsed in buffer and stabilized with a small amount of 2%
agarose in PBS to hold them together. They were then dehydrated through a graded
series of ethanol to 100%, followed by propylene oxide, 100%.
They were infiltrated with Epon resin (Ted Pella, Redding, CA) in a 1:1 solution
of Epon:propylene oxide overnight on a rocker at room temperature. The following
day they were placed in fresh Epon for several hours and then embedded in Epon
overnight at 60 C. Thin sections were cut on a Leica EM UC7 ultramicrotome,
collected on formvar-coated grids, stained with uranyl acetate and lead citrate
and examined in a JEOL JEM 1011 transmission electron microscope at 80 kV.
Images were collected using an AMT digital imaging system (Advanced Microscopy
Techniques, Danvers, MA). These methods are similar to previous descriptions of
extracellular particle and mitochondria detection in astrocyte cultures
12 (link).
+ Open protocol
+ Expand
10

Astrocyte Ultrastructural Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Rat cortical astrocytes or pellets from astrocyte-conditioned media were
fixed in 2.0% glutaraldehyde in 0.1 M sodium cacodylate buffer, pH 7.4
(Electron Microscopy Sciences, Hatfield, PA) for one hour at room temperature on
a rocker. They were rinsed in cacodylate buffer, gently scraped and pelleted and
post-fixed in 1.0% osmium tetroxide in cacodylate buffer for one hour on
ice. They were rinsed in buffer and stabilized with a small amount of 2%
agarose in PBS to hold them together. They were then dehydrated through a graded
series of ethanol to 100%, followed by propylene oxide, 100%.
They were infiltrated with Epon resin (Ted Pella, Redding, CA) in a 1:1 solution
of Epon:propylene oxide overnight on a rocker at room temperature. The following
day they were placed in fresh Epon for several hours and then embedded in Epon
overnight at 60 C. Thin sections were cut on a Leica EM UC7 ultramicrotome,
collected on formvar-coated grids, stained with uranyl acetate and lead citrate
and examined in a JEOL JEM 1011 transmission electron microscope at 80 kV.
Images were collected using an AMT digital imaging system (Advanced Microscopy
Techniques, Danvers, MA). These methods are similar to previous descriptions of
extracellular particle and mitochondria detection in astrocyte cultures
12 (link).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!