Semi-Automated Data Collection Software for Single Particle Work

Graphical user interface of EM-SPC: Definition of exposure positions (red borders) and focus positions (blue borders) with a single mouse click. Graphical user interface of EM-SPC: Definition of exposure positions (red borders) and focus positions (blue borders) with a single mouse click.
EM-SPC is designed to allow a quick and flexible determination of areas of interests followed by an automated acquisition of spotscan images under low-dose conditions.

ribosomeFor single particle reconstruction the acquisition of numerous (> 10 000) high quality cryo-electron micrographs under low-dose conditions is required, to determine the 3D structure of macromolecular complexes with high resolution. However, conventional manual data collection of large data sets is very time consuming and inefficient. On the other hand fully automated data collection can produce a vast amount of data which needs enormous time of post-processing.
EM-SPC increases the number of “good” particles per session by manually preselecting suitable areas on the grid followed by an automated acquisition & reduces post-processing time providing data for 3D reconstructions with sub nanometer resolution within a few days. EM-SPC is installed on a wide range of TEMs, e.g. FEI Tecnai12/20/30, Polara, Titan Krios, Hitachi H-9500, JEOL JEM-1230/1400/2010/2100/2200/3100/3200FSC, Philips CM100/120/200/300 & Zeiss 912/922, Libra120/200, Kronos.

  • Semi-automated acquisition of single particles under low-dose conditions
  • Acquisition of a tiled image of the whole specimen (grid map)
  • Selection modes for data collection and focussing: Manual for non-regular grids, AutoCenter with reference hole, AutoPick with reference hole and given variation limits
  • Automated acquisition of previous defined positions including eucentricity check, position and beam centering, focussing and drift check
  • Acquisition modes: Single Image, Focal/Triple Pair, Dose Series, Tiled Images for tilted specimens including dynamic focussing
  • Supports several structures for acquisition (rectangular, hexagonal and triangular)
  • Works on regular and non-regular support films (e.g. Quantifoil, Lacey film)
  • Typical acquisition rate is 5,000 cryo-images per 24 hours (drift rate dependent)
  • Requires computerized stage control and TEM remote control