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License: GPL v3 Downloads Documentation Status Last Commit

easymode

pretrained neural networks for cellular cryoET

Easymode is a collection of general pretrained neural networks for cellular cryo-electron tomography (cryoET). It provides single command line interface functions to handle feature detection: segmentation of cellular features, coordinate extraction, and data preprocessing. Inspired by and based upon MemBrain-seg.

Documentation | Model library | Tutorials

installation

conda create -n easymode python=3.10 cudatoolkit=11.2 cudnn=8.1 git -c conda-forge
conda activate easymode
pip install tensorflow==2.11.0 protobuf==3.20.3
pip install git+https://github.com/mgflast/easymode.git

For full installation instructions including CUDA setup, Ais, and Pom, see the installation guide.

quick start

# List available models
easymode list

# Segment features
easymode segment ribosome microtubule --data warp_tiltseries/reconstruction

# Extract particle coordinates
easymode pick ribosome --data segmented/ --output coordinates/ribosome --size 2000000 --spacing 300

# Reconstruct tomograms (requires WarpTools and AreTomo3)
easymode reconstruct --frames frames/ --mdocs mdocs/ --apix 1.56 --dose 4.6

# Denoise tomograms
easymode denoise --method n2n --mode direct --data warp_tiltseries/reconstruction --output denoised

Pretrained networks are hosted on HuggingFace and downloaded automatically on first use. See the user guide for detailed usage.

training data

All networks were trained on a dataset of over 2000 tilt series from 50+ sources, covering many prokaryotic, archaeal, and eukaryotic species, different sample preparation techniques, hardware configurations, pixel sizes, electron doses, and defocus values. Tomograms were reconstructed in multiple flavours using WarpTools, AreTomo3, cryoCARE, and DeepDeWedge at 10.0 A voxel size.

Not all training data sources are currently listed; a number of contributors prefer to remain anonymous so long as their work is not published. These contributions are marked with *.

ID Contributor / source Sample type N (annotated) Pixel size (A)
001_HELA Mart Last milled H. sapiens (HeLa) 60 (59) 1.51
002_U2OS Mart Last milled H. sapiens (U2OS) 40 (26) 2.15
003_HSPERM Tom Dendooven, Alia dos Santos, Matteo Allegretti milled H. sapiens (spermatozoa) 56 (41) 1.50
004_* * * 23 (20) 1.68
005_FIBRO Tom Hale milled H. sapiens (fibroblasts) 52 (47) 1.33
006_* * * 20 (16) 1.69
007_APOF EMPIAR-10491 purified apoferritin 37 (18) 0.79
008_HIV EMPIAR-10164 purified HIV particles 10 (4) 0.68
009_SCEREV Sebastian Tacke, Elisa Lisicki,
Tatjana Taubitz, Stefan Raunser
milled (hpf, pfib) S. cerevisiae 64 (51) 1.56
010_RIBO EMPIAR-11111 purified E. coli 70S ribosomes 25 (19) 1.07
011_CHLO EMPIAR-12612 milled S. oleracea chloroplasts 23 (18) 3.52
012_CHLAMY EMPIAR-11830 milled C. reinhardtii 52 (50) 1.96
013_DIAT EMPIAR-11747 milled T. pseudonana 7 (1) 1.07
014_CILIA EMPIAR-11078 milled C. reinhardtii ciliary base 23 (19) 3.42
015_MMVOLTA CDPDS-10452 whole M. mycoides cells 15 (15) 1.53
016_PHANTOM CDPDS-10440, CDPDS-10445 E. coli lysate with added proteins 19 (17) 1.53
017_MYCP EMPIAR-10499 whole M. pneunomiae cells 65 (27) 1.70
018_ECM EMPIAR-11897 lift-out H. sapiens (extracellular matrix) 39 (24) 2.14
019_ECOLI EMPIAR-12413 milled E. coli 44 (19) 1.90
020_* * * 30 (25) 2.13
021_* * * 8 (7) 3.02
022_SCOV EMPIAR-10493 purified SARS-CoV-2 virions 20 (12) 1.53
023_SPORE EMPIAR-12176 milled E. intestinalis 24 (11) 2.06
024_* * * 17 (6) 1.96
025_RPE EMPIAR-10989 cellular periphery H. sapiens (RPE1) 3 (3) 3.45
026_EHV EMPIAR-11896 Emiliania huxleyi virus 201 40 (10) 2.08
027_NUCFT Forson Gao milled S. cerevisiae nuclei 21 (15) 1.51
028_ROOF CDPDS-10434 cellular periphery H. sapiens (HEK293) 20 (19) 2.17
029_TKIV EMPIAR-11058 milled T. kivui 17 (7) 3.52
030_LDN Mart Last cellular periphery H. sapiens (U2OS) 26 (7) 2.74
031_MITO Mart Last milled H. sapiens (HeLa, mitochondria) 63 (59) 1.34
032_* * * 40 (24) 1.63
033_NPC EMPIAR-11830 (same source as 012_CHLAMY) milled C. reinhardtii (nuclear envelope) 36 (36) 1.96
034_DICTYO EMPIAR-11845 milled D. discoideum 152 (68) 2.18
035_GEM EMPIAR-11561 milled H. sapiens (HeLa, mitochondria) 15 (14) 3.43
036_MACRO EMPIAR-12457 milled H. sapiens (macrophages) 39 (21) 2.41
037_MESWT EMPIAR-12460 milled M. musculus (embryonic stem cell) 159 (26) 2.68
038_POMBE EMPIAR-10988 milled S. pombe 9 (6) 3.37
039_JUMBO EMPIAR-11198 milled E. amylovora + RAY phage 32 (4) 4.27
040_SLO CDPDS-10004 milled (hpf, pfib) C. elegans 100 (24) 1.50
041_RPEM Cong Yu milled H. sapiens (RPE1) 17 (7) 1.57
042_NPCSC EMPIAR-10466 milled S. cerevisiae 177 (0) 3.45
043_DICTY2 EMPIAR-11899 (to be included after validation) milled D. discoideum 0 (0) 1.22
044_JURKAT Mart Last milled H. sapiens (Jurkat) 177 (0) 1.97
045_NPHL * * 231 (0) 1.56
046_ROOF2 CDPDS-10431 cellular periphery H. sapiens (HEK293) 87 (0) 2.17
047_ECPP7 CDPDS-10455 E. coli + PP7 virus-like particles 10 (0) 1.50
048_ELSO CDPDS-10444 human endo-/lysosomes 10 (0) 1.54
049_CHR * * 46 1.97
050_MHSP EMPIAR-13145 milled H. sapiens (HeLa) 239 2.31
051_ASSC * milled S. cerevisiae 82 2.41

EMPIAR: EM Public Image Archive | CDPDS: CryoET Data Portal

references

  1. Tegunov et al., 'Multi-particle cryo-EM refinement with M visualizes ribosome-antibiotic complex at 3.5 A in cells', Nature Methods (2021): https://doi.org/10.1038/s41592-020-01054-7
  2. Tegunov & Cramer, 'Real-time cryo-electron microscopy data preprocessing with Warp', Nature Methods (2019): https://doi.org/10.1038/s41592-019-0580-y
  3. Peck et al., 'AreTomoLive: Automated reconstruction of comprehensively-corrected and denoised cryo-electron tomograms in real-time and at high throughput', bioRxiv (2025): https://doi.org/10.1101/2025.03.11.642690
  4. Lamm et al., 'MemBrain v2: an end-to-end tool for the analysis of membranes in cryo-electron tomography', bioRxiv (2024): https://doi.org/10.1101/2024.01.05.574336
  5. Burt et al., 'An image processing pipeline for electron cryo-tomography in RELION-5', FEBS Open Bio (2024): https://doi.org/10.1002/2211-5463.13873
  6. Buchholz et al., 'Content-aware image restoration for electron microscopy', IEEE (2019): https://doi.org/10.1109/ISBI.2019.8759519
  7. Wiedemann & Heckel, 'A deep learning method for simultaneous denoising and missing wedge reconstruction in cryogenic electron tomography', Nature Communications (2024): https://doi.org/10.1038/s41467-024-51438-y
  8. Last et al., 'Streamlining segmentation of cryo-electron tomography datasets with Ais', eLife (2024): https://doi.org/10.7554/eLife.98552.3

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