Program
Monday, March 3, 2025
09:00 – 09:30 | Registration |
09:30 – 09:45 | Opening of the course |
09:45 – 10:00 | Short introduction of participants |
10:00 – 10:45 | Sample preparation for biological transmission electron microscopy Andreas Nowak |
10:45 – 11:00 | Expected results from the room temperature methods – some examples Vlada Filimoněnko |
Coffee break | |
11:15 – 12:00 | Preparation of samples for TEM in cryogenic conditions Frédéric Leroux – remotely |
12:00 – 12:15 | Expected results from the cryo methods – some examples Vlada Filimoněnko |
12:15 – 12:45 | Core facility tour Dominik Pinkas |
Lunch | |
13:15 – 17:00 | Practical 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 – 09:45 | Sampe preparation methods for scanning electron microscopy Markéta Dalecká |
09:45 – 10:30 | Correlative workflows for cryo-electron microscopy: close-to-native imaging Martin Uher |
Coffee break | |
10:45 – 12:45 | Practical 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 – 17:30 | Practical 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 – 09:45 | Electron microscopy basics Oldřich Benada |
09:45 – 10:30 | Electron tomography in TEM and STEM – volume information with high resolution Dominik Pinkas |
Coffee break | |
10:45 – 12:45 | Practical 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 – 17:30 | Practical 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 – 09:45 | Principles of scanning transmission electron microscopy (STEM) and EDS elemental analysis Régis Ravelle Chapuis |
09:45 – 10:30 | EDS data collection, processing, and interpretation Régis Ravelle Chapuis |
Coffee break | |
10:45 – 12:45 | Practical 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 – 17:30 | Practical 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 – 09:45 | Volume EM approaches Markéta Dalecká |
09:45 – 10:30 | Preparation of digital images for publication Oldřich Benada |
Coffee break | |
10:45 – 12:45 | Practical 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 – 15:30 | Practical 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 – 16:10 | Round table: Application-oriented selection of EM techniques Vlada Filimoněnko |
16:10 – 16:25 | Evaluation of the course, closing remarks Vlada Filimoněnko |