Advanced EM - 3D imaging
This three-day workshop is intended for students/postdocs/staff who want to learn about TEM principles and some of the advanced TEM methods. The course will focus on electron tomography, cryo-single particle analysis (including the whole workflow of sample preparation, screening, data acquisition and processing), and negative staining method. The last day is dedicated to the introduction of the concepts of volume SEM methods including SBF-SEM, array tomography and consecutive image analysis. The course is limited to about 15 participants per each day.


Event Timing: Jun 11th-13th, 2024
Event Address: LEM, Biology Centre CAS, Branisovska 31
Contact us at holland@paru.cas.cz
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Day 1 - TEM
09:30 Welcome and workshop overview
10:00 Basics of TEM–Theory: basic principles, TEM construction, resolution, alignment, imaging & Electron tomography (T. Bílý)
11:30 Specimen preparation for TEM/tomography, examples of applications (M. Vancová)

12:30 Lunch

13:30 Practical session - basic operation TEM
Group 1: JEOL 1400 flash equipped with EMSIS XAROSA (T. Bílý)
Group 2: JEOL 2100F equipped with GATAN K2 direct camera (Z. Gardian)

15:30 Practical session - Electron tomography (T. Bílý)

18:00 Dinner

Day 2 - SPA
09:00 Introduction to Negative Stain/ Cryo-EM single particle analysis (Z. Gardian, E. Ďurinová)
11:30 How to prepare samples for SPA (L. Bumba)

12:30 Lunch

13:30 Practical session
Group 1: Negative staining TEM (T. Bílý, E. Ďurinová)
Group 2: Cryo-fixation and cryo-EM visualization of proteins, workflow(Z. Gardian)

15:30 Practical session – Single-particle image analysis

Day 3 - SBF & AT
09:00 Serial Block Face SEM and Array tomography - what we can visualize with these Volume SEM techniques (J. Týč)
10:45 Image processing and analysis of Volume EM data (F. Kitzberger)

12:00 Lunch

13:30 Practical session 1
Group 1: SBF-SEM, Apreo Volume Scope (J. Týč)
Group 2: Image processing (F. Kitzberger)

15:30 Practical session 2
Group 1: SBF-SEM, Apreo Volume Scope (J. Týč)

Group 2: Image processing (F. Kitzberger)

Morning lectures and afternoon practice also include coffee breaks.
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