Program

Monday, March 3, 2025

09:00 – 09:30Registration
09:30 – 09:45Opening of the course
09:45 – 10:00Short introduction of participants
10:00 – 10:45Sample preparation for biological transmission electron microscopy
Andreas Nowak
10:45 – 11:00Expected results from the room temperature methods – some examples
Vlada Filimoněnko
Coffee break
11:15 – 12:00Preparation of samples for TEM in cryogenic conditions
Frédéric Leroux – remotely
12:00 – 12:15Expected results from the cryo methods – some examples
Vlada Filimoněnko
12:15 – 12:45Core facility tour
Dominik Pinkas
Lunch
13:15 – 17:00Practical sessions (3 x 60 min + 10 min coffee break)
Routine sample preparation | Markéta Dalecká
Room temperature sectioning of resin-embedded samples | Andreas Nowak
Plunge-freezing – vitrification in thin layer|Dominik Pinkas

Tuesday, March 4, 2025

09:00 – 09:45Sampe preparation methods for scanning electron microscopy
Markéta Dalecká
09:45 – 10:30Correlative workflows for cryo-electron microscopy: close-to-native imaging
Martin Uher
Coffee break
10:45 – 12:45Practical sessions (1 x 120 min)
High-pressure freezing and freeze-substitution | Dominik Pinkas
Cryo-sectioning – Tokuyasu method for efficient immunolabeling | Andreas Nowak
Frozen-hydrated lamella preparation in cryo-FIB-SEM | Martin Uher
Lunch
13:30 – 17:30Practical sessions – continuation (2 x 120 min, incl. coffee break)
High-pressure freezing and freeze-substitution | Dominik Pinkas
Cryo-sectioning – Tokuyasu method for efficient immunolabeling | Andreas Nowak
Frozen-hydrated lamella preparation in cryo-FIB-SEM | Martin Uher
17:30 – ?Social event – sponsored by Specion

Wednesday, March 5, 2025

09:00 – 09:45Electron microscopy basics
Oldřich Benada
09:45 – 10:30Electron tomography in TEM and STEM – volume information with high resolution
Dominik Pinkas
Coffee break
10:45 – 12:45Practical sessions (1 x 120 min)
Freeze-fracture replica – a tool to study of membranes | Markéta Dalecká
Cryo-fluorescent light microscopy for CLEM | Martin Kopecký, Pavla Molínová
Cryo-electron tomography of FIB-milled lamella | Dominik Pinkas
Lunch
13:30 – 17:30Practical sessions – continuation (2 x 120 min, incl. coffee break)
Freeze-fracture replica – a tool to study of membranes | Markéta Dalecká
Cryo-fluorescent light microscopy for CLEM | Martin Kopecký, Pavla Molínová
Cryo-electron tomography of FIB-milled lamella | Dominik Pinkas

Thursday, March 6, 2025

09:00 – 09:45Principles of scanning transmission electron microscopy (STEM) and EDS elemental analysis
Régis Ravelle Chapuis
09:45 – 10:30EDS data collection, processing, and interpretation
Régis Ravelle Chapuis
Coffee break
10:45 – 12:45Practical sessions (1 x 120 min)
Studying surfaces and volumes using scanning electron microscope | Markéta Dalecká
Observation of prepared samples in routine TEM | Dominik Pinkas
Elemental mapping and analysis by STEM-EDS | Régis Ravelle Chapuis
Lunch
13:30 – 17:30Practical sessions – continuation (2 x 120 min, incl. coffee break)
Studying surfaces and volumes using scanning electron microscope | Markéta Dalecká
Observation of prepared samples in routine TEM | Dominik Pinkas
Elemental mapping and analysis by STEM-EDS | Régis Ravelle Chapuis

Friday, March 7, 2025

09:00 – 09:45Volume EM approaches
Markéta Dalecká
09:45 – 10:30Preparation of digital images for publication
Oldřich Benada
Coffee break
10:45 – 12:45Practical sessions (1 x 120 min)
Volume data acquisition from thick sections – STEM-tomography | Dominik Pinkas
Volume reconstruction and segmentation of 3D data | Markéta Dalecká
Lunch
13:30 – 15:30Practical sessions continuation (1 x 120 min)
Volume data acquisition from thick sections – STEM-tomography | Dominik Pinkas
Volume reconstruction and segmentation of 3D data | Markéta Dalecká
Coffee break
15:40 – 16:10Round table: Application-oriented selection of EM techniques
Vlada Filimoněnko
16:10 – 16:25Evaluation of the course, closing remarks
Vlada Filimoněnko