AAV1 or AAV5 particles and purified wt AAVR were mixed at a molar ratio of 1:120 (AAV:AAVR) at 4 °C for 6 h. An aliquot of 3 μl of each mixture was loaded onto a glow-discharged, carbon-coated copper grid (GIG, Au 1.2/1.3 200 mesh; Lantuo, China) bearing an ultrathin layer of carbon. The grid was then blotted for 4 s with a blot force of 0 in 100% relative humidity and plunge-frozen in liquid ethane using a Vitrobot Mark IV (FEI, USA). Cryo-EM data were collected with a 200 kV Arctica D683 electron microscope (FEI, USA) and a Falcon II direct electron detector (FEI, USA). A series of micrographs were collected as movies (19 frames, 1.2 s) and recorded with −2.2 to −0.5 μm defocus at a calibrated magnification of ×110,000, resulting in a pixel size of 0.93 Å per pixel. Statistics for data collection and refinement are summarized in Supplementary Table 1 .
Arctica d683 electron microscope
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Arctica D683 is an electron microscope designed for high-resolution imaging of biological samples. It utilizes a direct electron detection camera to capture images with exceptional detail and clarity.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using arctica d683 electron microscope
1
Cryo-EM Analysis of AAV-AAVR Complexes
2
Single-particle cryo-EM of AAV-AAVR complex
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AAV9 or AAVPHP.eB particles and purified wild-type AAVR were mixed at a molar ratio of 1:120 (AAV:AAVR) at 4°C for 1 h. An aliquot of 3 μL of each mixture was loaded onto a glow-discharged, carbon-coated copper grid (GIG, Au 2/1 200 mesh; Lantuo, China) bearing an ultrathin layer of carbon. The grid was then blotted for 4.5 s with a blot force of 0 in 100% relative humidity and plunge frozen in liquid ethane using a Vitrobot Mark IV (FEI, USA). Cryo-EM data were collected with a 200 kV Arctica D683 electron microscope (FEI, USA) and a Falcon II direct electron detector (FEI, USA). A series of micrographs were collected as movies (19 frames, 1.2 s) and recorded with −2.2 to −0.5-μm defocus at a calibrated magnification of 110,000×, resulting in a pixel size of 0.93 Å per pixel. Statistics for data collection and refinement are summarized in Table S1 .
3
Cryo-EM Analysis of AAV-AAVR Interaction
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AAV9 or AAV-PHP.eB particles and purified wt AAVR were mixed at a molar ratio of 1:120 (AAV:AAVR) at 4 °C for 1hr. An aliquot of 3 μl of each mixture was loaded onto a glow-discharged, carbon-coated copper grid (GIG, Au 2/1 200 preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this this version posted January 24, 2022. ; https://doi.org/10.1101/2022.01.23.477411 doi: bioRxiv preprint mesh; Lantuo, China) bearing an ultrathin layer of carbon. The grid was then blotted for 4.5 s with a blot force of 0 in 100% relative humidity and plungefrozen in liquid ethane using a Vitrobot Mark IV (FEI, USA). Cryo-EM data were collected with a 200 kV Arctica D683 electron microscope (FEI, USA) and a Falcon II direct electron detector (FEI, USA). A series of micrographs were collected as movies (19 frames, 1.2 s) and recorded with -2.2 to -0.5 μm defocus at a calibrated magnification of 110,000×, resulting in a pixel size of 0.93 Å per pixel. Statistics for data collection and refinement are summarized in Supplementary Table 1.
The copyright holder for this this version posted January 24, 2022. ; https://doi.org/10.1101/2022.01.23.477411 doi: bioRxiv preprint mesh; Lantuo, China) bearing an ultrathin layer of carbon. The grid was then blotted for 4.5 s with a blot force of 0 in 100% relative humidity and plungefrozen in liquid ethane using a Vitrobot Mark IV (FEI, USA). Cryo-EM data were collected with a 200 kV Arctica D683 electron microscope (FEI, USA) and a Falcon II direct electron detector (FEI, USA). A series of micrographs were collected as movies (19 frames, 1.2 s) and recorded with -2.2 to -0.5 μm defocus at a calibrated magnification of 110,000×, resulting in a pixel size of 0.93 Å per pixel. Statistics for data collection and refinement are summarized in Supplementary Table 1.
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