Course description:

This course provides an insight into the complexity and principles of sample preparation necessary for ultrastructural analysis by Transmission and Scanning electron microscopy. An emphasis is given to definition of critical steps in sample preparation workflows (from harvesting and fixation to contrasting, immunolabelling, resin embedding, ultrathin sectioning etc.) and their optimization to suit specific type of samples (from nanoparticles to cells, tissues and bio-composites) and to specific aims of the analysis. The practical part of the course will focus primarily on methods suitable for routine quality control and quantitative analysis of particulate and suspension samples including nanoparticles, biopolymers, viruses & bacteria, liposomes, exosomes etc. The EM-SPS course is organized by the Imaging Methods Core Facility at BIOCEV as an annual course supported by the National Infrastructure for Biological and Medical Imaging (Czech-BioImaging, Ministry of Education, Youth and Sports – Large Research Infrastructure, LM2023050).

• Day 1: Lectures, day
• Days 2-4: Demo and hands-on training

METHODS & WORKFLOWS (covered in lectures/demo/hands-on)

• Sampling and sample harvesting
• Negative staining and plunge freezing (for nanoparticles, protein complexes, liposomes)
• Chemical fixation, contrasting, dehydration and resin embedding (for cells and tissues)
• Semithin sectioning and section staining (for LM)
• Ultrathin sectioning and section contrasting (for TEM)
• Sample drying, mounting and sputter coating (for SEM)
• Fine structure immuno-gold labelling (Direct immuno-EM)
• TEM/SEM screening workflows for various types of samples

Registration and more information: