Instruct.SI CryoEM Advanced Workshop 2025, Single Particle Analysis
We are pleased to report the successful completion of the Instruct.SI CryoEM Advanced Workshop 2025, Single Particle Analysis, held from 11 to 14 November 2025 at the National Institute of Chemistry in Ljubljana. This hands-on workshop brought together a small but highly engaged group of six participants, creating an intensive learning environment focused on practical skills throughout the entire cryo-EM workflow. Participants presented their research projects as flash presentations, leading to a fruitful exchange of ideas among peers from different fields.
A key feature of this year’s workshop was the opportunity for participants to bring their own research samples. Four attendees took advantage of this, and all participants prepared multiple cryoEM grids during the practical sessions. Under close mentorship from several tutors, they successfully vitrified their samples, gained confidence in handling the Vitrobot IV, and developed a strong understanding of best practices for grid preparation. The small-group format allowed for individualised guidance and ensured that everyone produced usable grids and learned from real experimental outcomes. Beyond sample preparation, the workshop also emphasised microscope operation, screening strategies, and data collection. Participants worked with the 200 kV Glacios cryoTEM, learning step by step from how to load grids to setting up data collection.
The final part of the programme focused on data processing and 3D reconstruction, giving participants a complete overview of the single particle analysis pipeline – from motion correction and CTF estimation to particle picking, 2D classification, and initial 3D refinement. Guided sessions supported by five tutors enabled each participant to follow the process on a real dataset and understand how individual steps influence the final result.
Overall, the workshop offered a concentrated and highly practical experience, allowing participants not only to observe but also to actively perform each stage of the workflow. The positive feedback we received underscores the value of small, skill-focused workshops within the Instruct.SI community. Building on this success, we aim to continue offering advanced, practice-oriented training opportunities to support researchers in Slovenia as they further develop expertise in cryo-electron microscopy.
More information: Instruct.SI CryoEM Advanced Workshop 2025, Single Particle Analysis - Instruct.SI