Sub-pixel electron detection using a convolutional neural network

Ultramicroscopy, vol. 218, pp. 113091

Abstract

Modern direct electron detectors (DEDs) provided a giant leap in the use of cryogenic electron microscopy (cryo-EM) to study the structures of macromolecules and complexes thereof. However, the currently available commercial DEDs, all based on the monolithic active pixel sensor, still require relative long exposure times and their best results have only been obtained at 300 keV. There is a need for pixelated electron counting detectors that can be operated at a broader range of energies, at higher throughput and higher dynamic range. Hybrid Pixel Detectors (HPDs) of the Medipix family were reported to be unsuitable for cryo-EM at energies above 80 keV as those electrons would affect too many pixels. Here we show that the Timepix3, part of the Medipix family, can be used for cryo-EM applications at higher energies. We tested Timepix3 detectors on a 200 keV FEI Tecnai Arctica microscope and a 300 keV FEI Tecnai G2 Polara microscope. A correction method was developed to correct for per-pixel differences in output. Timepix3 data were simulated for individual electron events using the package Geant4Medipix. Global statistical characteristics of the simulated detector response were in good agreement with experimental results. A convolutional neural network (CNN) was trained using the simulated data to predict the incident position of the electron within a pixel cluster. After training, the CNN predicted, on average, 0.50 pixel and 0.68 pixel from the incident electron position for 200 keV and 300 keV electrons respectively. The CNN improved the MTF of experimental data at half Nyquist from 0.39 to 0.70 at 200 keV, and from 0.06 to 0.65 at 300 keV respectively. We illustrate that the useful dose-lifetime of a protein can be measured within a 1 second exposure using Timepix3.

Authors 9

1. J. Paul van Schayck

   Maastricht University

   Affiliation as printed

   Maastricht MultiModal Molecular Imaging Institute (M4I), FHML, Maastricht University, Maastricht, The Netherlands

2. Eric van Genderen

   Paul Scherrer Institute

   Affiliation as printed

   BIO, Laboratory for Nanoscale Biology. Paul Scherrer Institut, Villigen CH-5232, Switzerland

3. E. Maddox Aachen

   Leiden University · Amsterdam Scientific Instruments (Netherlands)

   Affiliation as printed

   Leiden University, Leiden, The Netherlands; Amsterdam Scientific Instruments (ASI), Amsterdam, The Netherlands

   Amsterdam Scientific Instruments (ASI), Amsterdam, The Netherlands

   Leiden University, Leiden, The Netherlands

4. Lucas Roussel

   Maastricht University

   Affiliation as printed

   Maastricht MultiModal Molecular Imaging Institute (M4I), FHML, Maastricht University, Maastricht, The Netherlands

5. Hugo Boulanger

   Maastricht University

   Affiliation as printed

   Maastricht MultiModal Molecular Imaging Institute (M4I), FHML, Maastricht University, Maastricht, The Netherlands

6. E. Fröjdh

   Paul Scherrer Institute

   Affiliation as printed

   PSD Detector Group, Paul Scherrer Institute, Villigen CH-5232, Switzerland

7. Jan Pieter Abrahams Aachen

   Leiden University · University of Basel · Paul Scherrer Institute

   Affiliation as printed

   Leiden University, Leiden, The Netherlands; BIO, Laboratory for Nanoscale Biology. Paul Scherrer Institut, Villigen CH-5232, Switzerland; Center for Cellular Imaging and NanoAnalytics, Biozentrum, University of Basel, Basel CH-4058, Switzerland

   BIO, Laboratory for Nanoscale Biology. Paul Scherrer Institut, Villigen CH-5232, Switzerland

   Center for Cellular Imaging and NanoAnalytics, Biozentrum, University of Basel, Basel CH-4058, Switzerland

   Leiden University, Leiden, The Netherlands

8. Peter J. Peters

   Maastricht University

   Affiliation as printed

   Maastricht MultiModal Molecular Imaging Institute (M4I), FHML, Maastricht University, Maastricht, The Netherlands

9. Raimond B. G. Ravelli corresponding

   Maastricht University

   Affiliation as printed

   Maastricht MultiModal Molecular Imaging Institute (M4I), FHML, Maastricht University, Maastricht, The Netherlands. Electronic address: rbg.ravelli@maastrichtuniversity.nl

   Maastricht MultiModal Molecular Imaging Institute (M4I), FHML, Maastricht University, Maastricht, The Netherlands