Perforated carbon film-covered microscopical 200 mesh grids (R1/4 batch of Quantifoil, MicroTools GmbH, Jena, Germany) were cleaned with chloroform and hydrophilized by 60 s glow discharging at 10 mA in a Safematic CCU-010 device (Safematic GmbH, Zizers, Switzerland). Subsequently, 4 L aliquots of the sample solution were applied to the grids. The samples were vitrified by automatic blotting and plunge freezing with an FEI Vitrobot Mark IV (Thermo Fisher Scientific Inc., Waltham, MA, USA) using liquid ethane as a cryogen. The vitrified specimens were transferred to the autoloader of an FEI TALOS ARCTICA electron microscope (Thermo Fisher Scientific Inc., Waltham, MA, USA). This microscope is equipped with a high-brightness field-emission gun (XFEG) operated at an acceleration voltage of 200 kV. Micrographs were acquired on an FEI Falcon 3 direct electron detector (Thermo Fisher Scientific Inc., Waltham, MA, USA) using a 100 μm objective aperture. Graphical analysis of the images was performed using Image J v1.53k.
Fei falcon 3 direct electron detector
Manufactured by Thermo Fisher Scientific
Sourced in United States
The FEI Falcon 3 is a direct electron detector designed for use in transmission electron microscopy (TEM). It captures high-resolution images by directly detecting the electrons transmitted through the sample, without the need for scintillators or optical components. The Falcon 3 provides improved signal-to-noise ratio and spatial resolution compared to conventional charge-coupled device (CCD) or complementary metal-oxide-semiconductor (CMOS) detectors.
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Lab products found in correlation
4 protocols using fei falcon 3 direct electron detector
1
Cryo-EM Sample Preparation Protocol
2
Cryogenic Electron Microscopy Sample Preparation
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Perforated carbon film-covered microscopical 200 mesh grids (R1/4 batch of Quantifoil, MicroTools GmbH, Jena, Germany) were cleaned with chloroform and hydrophilized by 60 s glow discharging at 10 mA in a Safematic CCU-010 device (safematic GmbH, Zizers, Switzerland). Subsequently, 4 μL aliquots of the sample solution were applied to the grids. The samples were vitrified by automatic blotting and plunge freezing with a FEI Vitrobot Mark IV (Thermo Fisher Scientific Inc., Waltham, MA, USA) using liquid ethane as cryogen.
The vitrified specimens were transferred to the autoloader of a FEI TALOS ARCTICA electron microscope (Thermo Fisher Scientific Inc., Waltham, MA, USA). This microscope is equipped with a high-brightness field-emission gun (XFEG) operated at an acceleration voltage of 200 kV. Micrographs were acquired on a FEI Falcon 3 direct electron detector (Thermo Fisher Scientific Inc., Waltham, MA, USA) using a 100 μm objective aperture.
The vitrified specimens were transferred to the autoloader of a FEI TALOS ARCTICA electron microscope (Thermo Fisher Scientific Inc., Waltham, MA, USA). This microscope is equipped with a high-brightness field-emission gun (XFEG) operated at an acceleration voltage of 200 kV. Micrographs were acquired on a FEI Falcon 3 direct electron detector (Thermo Fisher Scientific Inc., Waltham, MA, USA) using a 100 μm objective aperture.
3
Cryo-Electron Tomography of Vitrified Specimens
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Vitrified specimens (see above) intended for cryo‐ET were transferred to the autoloader of an FEI TALOS ARCTICA electron microscope (Thermo Fisher Scientific Inc., Waltham, MA, USA). This microscope is equipped with a high‐brightness field‐emission gun (XFEG) operated at an acceleration voltage of 200 kV. Micrographs were acquired on an FEI Falcon 3 direct electron detector (Thermo Fisher Scientific Inc., Waltham, MA, USA) at a nominal magnification of 28 000×, corresponding to a calibrated pixel size of 3.64 Å per pixel.
Tomography series were recorded in the context of FEI Tomography Software V 4.3.1. The 4096×4096 pixel images were recorded in the tilt angle range of ±65° in 2° increments, with a total electron dose of 180 e Å−2. The 3D volume reconstructions were calculated with the help of INSPECT3D Software V4.4 (Thermo Fisher Scientific Inc., Waltham, MA, USA) and visualised with Imod V4.9.10.27
Tomography series were recorded in the context of FEI Tomography Software V 4.3.1. The 4096×4096 pixel images were recorded in the tilt angle range of ±65° in 2° increments, with a total electron dose of 180 e Å−2. The 3D volume reconstructions were calculated with the help of INSPECT3D Software V4.4 (Thermo Fisher Scientific Inc., Waltham, MA, USA) and visualised with Imod V4.9.10.
4
Qβ Phage Capsid-Influenza Virus Binding Control
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To control the binding of the
Qβ phage capsids to influenza virus by cryoTEM, Qβ[Sia1(98%),TAMRA(2%)]
(9a ) (concentration 200 nmol/L) was incubated with 15
μL of IAV X31 (1.2 mg/mL total protein) with gentle agitation
for 30 min at room temperature. A perforated (1 μm hole diameter)
carbon film-covered microscopical 200 mesh grid (R1/4 batch of Quantifoil,
MicroTools GmbH, Jena, Germany) was cleaned with chloroform and hydrophilized
by 60 s glow discharging before a 5 μL aliquot of the sample
solution was applied to the grid. The sample was vitrified by automatic
blotting and plunge freezing with a FEI Vitrobot Mark IV (ThermoFisher
Scientific Inc., Waltham, MA) using liquid ethane as the cryogen.
The vitrified specimen was transferred to the autoloader of a FEI
TALOS ARCTICA electron microscope (ThermoFisher Scientific Inc.) operated
at an acceleration voltage of 200 kV. Micrographs were acquired on
a FEI Falcon 3 direct electron detector (ThermoFisher Scientific Inc.)
using a 100 μm objective aperture and a nominal magnification
of 28,000× corresponding to a calibrated pixel size of 3.69 Å/pixel.
Qβ phage capsids to influenza virus by cryoTEM, Qβ[Sia1(98%),TAMRA(2%)]
(
μL of IAV X31 (1.2 mg/mL total protein) with gentle agitation
for 30 min at room temperature. A perforated (1 μm hole diameter)
carbon film-covered microscopical 200 mesh grid (R1/4 batch of Quantifoil,
MicroTools GmbH, Jena, Germany) was cleaned with chloroform and hydrophilized
by 60 s glow discharging before a 5 μL aliquot of the sample
solution was applied to the grid. The sample was vitrified by automatic
blotting and plunge freezing with a FEI Vitrobot Mark IV (ThermoFisher
Scientific Inc., Waltham, MA) using liquid ethane as the cryogen.
The vitrified specimen was transferred to the autoloader of a FEI
TALOS ARCTICA electron microscope (ThermoFisher Scientific Inc.) operated
at an acceleration voltage of 200 kV. Micrographs were acquired on
a FEI Falcon 3 direct electron detector (ThermoFisher Scientific Inc.)
using a 100 μm objective aperture and a nominal magnification
of 28,000× corresponding to a calibrated pixel size of 3.69 Å/pixel.
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