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Title | An electron counting algorithm improves imaging of proteins with low-acceleration-voltage cryo-electron microscope. |
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Journal, issue, pages | Commun Biol, Vol. 5, Issue 1, Page 321, Year 2022 |
Publish date | Apr 6, 2022 |
Authors | Dongjie Zhu / Huigang Shi / Chunling Wu / Xinzheng Zhang / |
PubMed Abstract | Relative to the 300-kV accelerating field, electrons accelerated under lower voltages are potentially scattered more strongly. Lowering the accelerate voltage has been suggested to enhance the signal- ...Relative to the 300-kV accelerating field, electrons accelerated under lower voltages are potentially scattered more strongly. Lowering the accelerate voltage has been suggested to enhance the signal-to-noise ratio (SNR) of cryo-electron microscopy (cryo-EM) images of small-molecular-weight proteins (<100 kD). However, the detection efficient of current Direct Detection Devices (DDDs) and temporal coherence of cryo-EM decrease at lower voltage, leading to loss of SNR. Here, we present an electron counting algorithm to improve the detection of low-energy electrons. The counting algorithm increased the SNR of 120-kV and 200-kV cryo-EM image from a Falcon III camera by 8%, 20% at half the Nyquist frequency and 21%, 80% at Nyquist frequency, respectively, resulting in a considerable improvement in resolution of 3D reconstructions. Our results indicate that with further improved temporal coherence and a dedicated designed camera, a 120-kV cryo-electron microscope has potential to match the 300-kV microscope at imaging small proteins. |
External links | Commun Biol / PubMed:35388174 / PubMed Central |
Methods | EM (single particle) |
Resolution | 2.51 - 4.35 Å |
Structure data | EMDB-32600: Structure of Coxsackievirus A10 for critical dose measurement at 120 kV EMDB-32601: Structure of Coxsackievirus A10 for critical dose measurement at 160 kV EMDB-32602: Structure of Coxsackievirus A10 for critical dose measurement at 300 kV EMDB-32603: Structure of Coxsackievirus A10 with Hybrid electron counting at 120 kV EMDB-32604: Structure of Coxsackievirus A10 with MCF electron counting at 120 kV EMDB-32605: Structure of Coxsackievirus A10 with WPF electron counting at 120 kV EMDB-32606: Structure of Coxsackievirus A10 with MCF electron counting and large-sized clusters at 120 kV EMDB-32607: Structure of Coxsackievirus A10 with PVF electron counting and large-sized clusters at 120 kV EMDB-32608: Structure of Coxsackievirus A10 with PCA electron counting and large-sized clusters at 120 kV EMDB-32609: Structure of Coxsackievirus A10 with MCF electron counting at 200 kV EMDB-32610: Structure of apo-ferritin with PCA electron counting at 300 kV EMDB-32611: Structure of Coxsackievirus A10 with Hybrid electron counting at 200 kV EMDB-32612: Structure of apo-ferritin with Hybrid electron counting at 120 kV EMDB-32613: Structure of apo-ferritin with MCF electron counting at 120 kV |
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