The established protocol of Bachurski et al. [50 (link)] was applied. Briefly, 5 µL of each sample was loaded onto formvar-coated copper grids (Science Services, Munich, Germany). After 20 min incubation, the grid-bound EVs were fixed with 2% paraformaldehyde for 5 min. Samples were washed with PBS and fixed again with 1% glutaraldehyde for 5 min, washed with Milli-Q water, and incubated with contrast dye (1.5% uranyl acetate) for 4 min. Images were acquired using a Gatan OneView 4 K camera (Gatan, Pleasanton, CA, USA) mounted on a Jem-2100Plus microscope (JEOL) operating at 200 kV.
Formvar coated copper grids
Manufactured by Science Services
Sourced in Germany
Formvar-coated copper grids are a type of laboratory equipment used for sample preparation in electron microscopy. They provide a stable and uniform support for thin samples, allowing for high-resolution imaging and analysis.
Automatically generated - may contain errors
6 protocols using formvar coated copper grids
1
Transmission Electron Microscopy of EVs
2
Transmission Electron Microscopy of Extracellular Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TEM was conducted based on the protocol previously described by Bachurski et al. [49 (link)]. Briefly, after loading 5 µL of an EV sample onto formvar-coated copper grids (Science Services, Munich, Germany), the EVs were fixed with 2% paraformaldehyde for 5 min, washed with PBS, fixed again for 5 min with 1% glutaraldehyde, washed with ddH2O, and incubated with contrast dye (1.5% uranyl acetate) for 4 min. Images were captured with a Gatan OneView 4K camera (Gatan, Pleasanton, CA, USA) on a Jem-2100Plus microscope (JEOL) operating at 200 kV.
3
Transmission Electron Microscopy of lEVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For transmission electron microscopic imaging, lEVs were harvested from one 10-cm dish (60 cm2 growth area), as described above. After isolation, the lEVs were resuspended in 10 µl PBS with Protease Inhibitor Cocktail (Roche, Basel, Switzerland, Cat# 4,693,132,001). 5 µl of the sample were loaded on Formvar-coated copper grids (Science Services, München, Germany) and fixed twice with 2% paraformaldehyde for 5 min and subsequently for 5 min with 1% glutaraldehyde. After extensive washing with distilled water, the sample was contrasted with 1.5% uranyl acetate for 4 min. Image acquisition was performed with a Jem-2100Plus (Jeol), which was operated at 200 kV, together with a Gatan OneView 4 K camera.
4
Transmission Electron Microscopy of Extracellular Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formvar-coated copper grids (Science Services, München) were loaded with 5 µl of diluted sample (1:20) containing 0.17 µg (Serum 10k xg), 0.24 µg (L-540 10k xg), 0.35 µg (L-540 100k xg) or 0.78 µg protein (Serum 100k xg) as determined by BCA protein assay. The grids and samples were incubated for 20 min before being fixed with 2% paraformaldehyde for 5 min. Samples were washed with PBS and fixed again with 1% glutaraldehyde for 5 min, washed with Milli-Q water and contrasted for 4 min with 1.5% uranyl acetate. Images were acquired using a Gatan OneView 4K camera mounted on a Jem-2100Plus (Jeol) operating at 200kV. Samples were analysed with ImageJ software (NIH).
5
Electron Microscopic Imaging of Plasma lEVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Electron microscopic imaging was performed as previously reported [36 (link)]. In brief, lEVs isolated from 1 ml plasma were finally resuspended in 10 µL PBS with Protease Inhibitor Cocktail (Roche, Cat# 4693132001). Prior to loading, the lEVs were diluted further 1:250 in PBS and 5 µl of the suspension were loaded on Formvar-coated copper grids (Science Services, München). After incubation for 20 min at RT, the sample was then fixed for 5 min with 2% paraformaldehyde. Subsequently, the EVs were washed with PBS and fixed again for 5 min with 1% glutaraldehyde. Finally, the sample was contrast stained with 1.5% uranyl acetate for 4 min. Image acquisition was performed with a Jem-2100Plus (Jeol) operating at 200 kV using a Gatan OneView 4 K camera.
6
Transmission Electron Microscopy Imaging of Extracellular Vesicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An established protocol was applied. 16 Briefly, 5 μL EV sample was loaded onto formvar-coated copper grids (Science Services, Munich, Germany) and incubated for 20 minutes. Then, the EVs were fixed with 2% paraformaldehyde for 5 minutes, washed with PBS, fixed again for 5 minutes with 1% glutaraldehyde, washed with ddH 2 O and incubated with contrast dye (1.5% uranyl acetate) for 4 minutes.
Images were taken with a Gatan OneView 4 K camera (Gatan, Pleasanton, California) mounted on a Jem-2100Plus microscope (JEOL) operating at 200 kV.
Images were taken with a Gatan OneView 4 K camera (Gatan, Pleasanton, California) mounted on a Jem-2100Plus microscope (JEOL) operating at 200 kV.
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
Revolutionizing how scientists
search and build protocols!