The extracted chromatin samples with a concentration of 50 ng/μl were used for negative stained electron tomography data collection. 10 μl of chromatin sample was mixed with 10 μl of 16 nm gold nanoparticle solution. An aliquot of 4 μl of the mixture was adsorbed onto the glow-discharged 200 mesh carbon continuous grids (Ted Pella, INC, CA, USA) for 1 min, manually blotted by putting a piece of #1 filter paper on the edge of the grid, and stained with 4 μl of uranyl acetate (UA, 2%) for 10s. After removing the UA, the grids were briefly washed with 4 μl water and further air dried. The grids were transferred into an FEI T20 electron microscope (FEI, Eindhoven) equipped with a LaB6 filament and operated at 200 kV. Negative stained micrographs of chromatin samples were obtained using a Gatan camera at different magnification according to the size of different chromatin fibers.
Fei t20 electron microscope
Manufactured by Thermo Fisher Scientific
The FEI T20 is a transmission electron microscope designed for high-resolution imaging and analysis of materials at the nanoscale. It features a 200 kV accelerating voltage, advanced optics, and a high-performance digital camera system for capturing detailed micrographs.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using fei t20 electron microscope
1
Negative Stained Electron Tomography of Chromatin
2
Structural analysis of GPC2-CT3 Fab complex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The CT3 antigen-binding fragment (Fab) was prepared using a Fab preparation kit (Thermo Scientific). GPC2 protein was mixed with CT3 Fab at 1:1 molar ratio in PBS. A 3 μl aliquot containing ∼0.01 mg/ml of the sample was applied for 20 s onto a carbon-coated 200 Cu mesh grid (Electron Microscopy Sciences, Protochips, Inc.) that had been glow discharged at 30 mA for 30 s (Pelco easiGlow, Ted Pella, Inc.), then negatively stained with 0.7% (w/v) uranyl formate for 40 s. Data was collected using a Tecnai FEI T20 electron microscope operating at 200 kV, with an electron dose of ∼40 e-/Å2 and a magnification of 100,000 x that resulted in a pixel size of 2.19 Å at the specimen plane. Images were acquired with an Eagle 2kx2k CCD camera (FEI) using a nominal defocus of 1100 nm and the SerialEM software.41 (link)
3
Chromatin Visualization via Gold Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isolated native chromatin (∼50 ng μL−1) was briefly incubated with gold nanoparticles before preparing positive staining electron microscopy grids. An aliquot of the mixture was adsorbed onto glow-discharged 200 mesh continuous carbon coated TEM grids (Ted Pella, INC, CA, USA) for 1 min, manually blotted by placing a piece of filter paper on the edge of the grid, and stained with 4 μL of uranyl acetate (UA, 2%) for 10 s. After removing the UA, the grids were briefly washed and air dried. The grids were transferred into a FEI T20 electron microscope (FEI, Eindhoven) equipped with LaB6 filament and operated at 200 kV. Positively stained micrographs of chromatin samples were obtained using a Gatan CCD camera at magnifications in accordance to the size of different chromatin fibers.
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!