The morphology of polymersomes was visualized by cryoTEM. The samples were prepared using the freeze-plunging method. Briefly, a small droplet of polymersome suspension (ca. 6 μl) was placed on a holey carbon film supported on a TEM copper grid (Electron Microscopy Sciences, Hatfield, PA), which was glow-charged in air before use. The excess solution was removed by blotting with a slip of filter paper (Whatman, Pittsburgh, PA) to form a thin liquid film on the grid. The specimen was then rapidly frozen by plunging into liquid ethane and stored in liquid nitrogen until it was transferred to a cryo-holder (Gatan 626) for examination via a Tecnai G2 F20 electron microscope (FEI Co., Hillsboro, OR). The TEM was operated at an acceleration voltage of 200 keV and images were recorded under a low-dose condition at a magnification of 25,000 or 50,000 by a CCD camera system (4k Eagle Camera, FEI). During the sample examination, the temperature of cryoholder was maintained at −179°C to prevent sublimation of vitreous water.
Tem copper grid
Manufactured by Electron Microscopy Sciences
Sourced in United States
The TEM copper grid is a specimen holder used in transmission electron microscopy (TEM) to support and secure thin samples for high-resolution imaging. Constructed from copper, it provides a stable platform for the specimen during the imaging process.
Automatically generated - may contain errors
Lab products found in correlation
Filter paper, by Cytiva (1 mentions)
Tecnai g2 f20 electron, by Thermo Fisher Scientific (1 mentions)
4k eagle camera, by Thermo Fisher Scientific (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Jem 2100f, by JEOL (1 mentions)
Eagle bottom mount camera, by Thermo Fisher Scientific (1 mentions)
Tecnai 12 twin transmission electron microscope, by Thermo Fisher Scientific (1 mentions)
4 protocols using tem copper grid
1
Cryogenic TEM Visualization of Polymersomes
2
Cryogenic TEM Imaging of Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cryogenic TEM imaging was
performed on the
FEI Tecnai 12 TWIN Transmission Electron Microscope, operating at
100 kV. Gels were immediately diluted five times with water to reduce
their viscosity and 6 μL of sample solution was placed on a
holey carbon film supported on a TEM copper grid (Electron Microscopy
Services, Hatfield, PA). All the TEM grids used for cryo-TEM imaging
were treated with plasma air to render the lacey carbon film hydrophilic.
A thin film of the sample solution was produced using the Vitrobot
with a controlled humidity chamber (FEI). After loading of the sample
solution, the lacey carbon grid was blotted using preset parameters
and plunged instantly into a liquid ethane reservoir precooled by
liquid nitrogen. The vitrified samples were then transferred to a
cryo-holder and cryo-transfer stage, which was cooled by liquid nitrogen.
To prevent sublimation of vitreous water, the cryo-holder temperature
was maintained below −170 °C during the imaging process.
All images were recorded by a SIS Megaview III wide-angle CCD camera.
performed on the
FEI Tecnai 12 TWIN Transmission Electron Microscope, operating at
100 kV. Gels were immediately diluted five times with water to reduce
their viscosity and 6 μL of sample solution was placed on a
holey carbon film supported on a TEM copper grid (Electron Microscopy
Services, Hatfield, PA). All the TEM grids used for cryo-TEM imaging
were treated with plasma air to render the lacey carbon film hydrophilic.
A thin film of the sample solution was produced using the Vitrobot
with a controlled humidity chamber (FEI). After loading of the sample
solution, the lacey carbon grid was blotted using preset parameters
and plunged instantly into a liquid ethane reservoir precooled by
liquid nitrogen. The vitrified samples were then transferred to a
cryo-holder and cryo-transfer stage, which was cooled by liquid nitrogen.
To prevent sublimation of vitreous water, the cryo-holder temperature
was maintained below −170 °C during the imaging process.
All images were recorded by a SIS Megaview III wide-angle CCD camera.
3
Characterization of Magnetic Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The particles were collected from solution using a permanent magnet and the supernatant was removed. Afterwards, particles were repeatedly washed with water and separated using a permanent magnet until the supernatant remained clear. The remaining particles were freeze-dried overnight. The pure particles were resuspended in water at a concentration of 0.1 mg/mL and were subsequently transferred onto a transmission election microscopy (TEM) copper grid (Electron Microscopy Science, USA) by repeatedly dipping the grid in the previously prepared solution. TEM images were taken using a JEM-2100F (JEOL, Japan) and particle sizes were analyzed using Image J (National Institute of Health, USA). For all conditions, 100 particles were measured to determine the resulting size distribution. Zeta potentials of the particles suspended in DI water were determined using a Zetasizer Nano ZS (Malvern Panalytical, United Kingdom).
4
Cryo-TEM Sample Preparation Protocol
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!