EMAN2 Introductory Tutorials

Please note that we are continuously developing and updating the tutorials. Most tutorials include the necessary data. If you have a request for a particular tutorial please feel free to email sludtke@bcm.edu .

Video tutorials and discussions

• EMAN2 Video Tutorials - These are a valuable supplement to the tutorials here, but are NOT a replacement

Standard Tutorials

Tutorial Data Downloads

The actual tutorials are in the sections below, this section contains the various test data sets.

Individual tutorial data downloads are available below, but for convenience, we have combined the data for the GUI introduction, single particle analysis tutorial, single particle 2-D dynamics tutorial and small tomography tutorial into a single download:

• http://blake.bcm.edu/dl/eman2_tutorials_all.zip

Alternatively, these individual data downloads cover the following standard workshop tutorial sequence:

• eman2_samples.zip

• Small Tomography Tutorial (for low end machines/laptops)

• Large Tomography Tutorial (for standard workstations)

• Single Particle Tutorial Data

• Practice data for 2-D variability analysis

Short tutorials and info sheets

• EMAN2/Jupyter - Using EMAN2 widgets from JupyterLab notebooks! (NEW - 2022)

• EMAN2/NegativeStain - special issues that impact processing negative stain single particle images

• Information about images

• Filtering and basic image processing

• CTF Simulator

Single particle analysis (SPA) - 2D particles

• Quick Introduction to the EMAN2 GUI (Start here if you have never used EMAN2!)

  • eman2_samples.zip

• Single particle analysis tutorial

  • Tutorial Data Download

• Particle picking tutorial (basic tutorial only)

• tutorial for the Neural network particle picker (requires CUDA for good performance)

• 2-D variability analysis tutorial

  • Practice data for 2-D variability analysis

• These haven't been updated in a while, please ask if you have problems

  • EMAN1 to EMAN2 Transition Guide - Also with useful quickstart guides for users of other software

  • Tilt Validation tutorial

  • Random Conical Tilt Tutorial

Tomography - tilt series through subtomogram averaging

• Complete workflow for tomography and subtomogram averaging (includes both small version (laptop ~12 Å) and large version (subnanometer on workstation))

  • Tomography and Subtomogram Averaging Tutorial

  • Small Data Download

  • Large Data Download

• Automated Cellular Tomogram Annotation using Neural Networks

  • Tomogram Annotation

• More advanced topics for subtomogram averaging

  • Advanced SPT methods - Latest (2023) workflow for in situ SPT.

  • Working with pseudosymmetric particles - Improving resolution and exploring variability in symmetric particles

  • Miscellaneous tools - Various SPT tools that may not be included in the main workflows.

Gaussian Mixture Model - for SPA and SPT data

• Heterogeneity analysis - Strongly suggest reading the paper (PMC8363932). While there have been some very significant improvements, it lays a foundation. This project has also bifurcated, as explained in the first tutorial below.

  • GMM GUI Tutorial - supports both SPA and SPT projects - suggest starting here for most users

  • Original GMM tutorial accompanying the 2021 paper

• Orientation refinement - Improve resolution for flexible structures. For detailed methods, see the 2024 paper.

  • GMM-based orientation refinement for SPA

  • A brief tutorial on GMM based orientation refinement for SPT

• Atomic model refinement - both single model and continuous movement.

Exchanging Data Between EMAN2 and Other Software (a bit outdated, untested with newer Relion versions)

• 2D Class Averaging with Relion

• 3D Refinement with Relion

• 3D Refinement with FreAlign

Archive of Workshop Material (Lectures, specific tutorials, etc.

• 2019 May UTMB Workshop (full site with talks)

  • Latest EMAN2.3 single particle tutorial PDF

  • Tutorial Data Download

  • Practice data for 2-D variability analysis

• 2019 January Tutorial for EMAN2.22

  • EMAN2-22Tutorial.pdf (version 1)

    • Tutorial Data Download

• 2018 May Tutorial for EMAN2.21a

  • EMAN2-21Tutorial.pdf (version 1)

    • Tutorial Data Download

• 2017 February Tutorial for EMAN2.2

  • EMAN2-2Tutorial.pdf

    • Tutorial Data Download

• 2016 Summer Tutorial for EMAN2.12 June 2016 snapshots

  • EMAN2-13_pre_Tutorial.pdf

    • Tutorial Data Download

  • EMAN2-13_pre_Heterogeneity.pdf

  • see also: http://www.sciencedirect.com/science/article/pii/S0076687916300362

• 2015 Fall Tutorial for EMAN2.12

  • EMAN2-12_Tutorial.pdf

  • Required data - Bgal.zip

  • EMAN2-12_Tutorial2_heterogeneity.pdf

  • Required data - ribosome.zip

• 2015 Workshop Tutorials and Demo data - This includes updated tutorials on both high resolution single particle analysis and 2D/3D heterogeneity analysis along with sample data, freshly updated for the EMAN2.12 release. The workshop also included a number of other related topics also available on that page.

• Single particle tomography - Tutorial for deprecated Single particle tomography (SPT) workflow.

• Tutorial from the 2014 Workshops in Shanghai, Brisbane and online - Based on EMAN2.1 beta4. Full single particle reconstruction tutorial

• Tutorial from the 2013 Workshop in Beijing (Data on same page) - Based on EMAN2.1alpha. Full single particle reconstruction tutorial

• Tutorial from the 2012 Workshop in Singapore (Data on same page) - Based on EMAN2.06. Full single particle reconstruction tutorial

• Tutorial from the 2011 EMAN2 workshop (Data on same page) - Based on EMAN2.01. Full single particle reconstruction tutorial

• Single particle reconstruction tutorial from 2008 Workshop ( - Full tutorial on single particle reconstruction using the workflow (uses Workshop demo data DVD)