Practical Course

This course provides participants with an insight into the complexity and principles of biological sample preparation necessary for routine ultrastructural analysis by Transmission or Scanning electron microscopy. An emphasis is given to definition of critical steps in sample preparation workflows 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 cover sample harvesting, chemical fixation, contrasting, resin embedding, ultra-thin sectioning, negative staining, critical point drying, plunge freezing and more. 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).

VENUE:

Imaging methods core facility at BIOCEV, Průmyslová 595, Vestec, Prague-West, 252 50, Czech Republic

DATE:

April 22-25, 2024 (4 days): (day 1: Lectures, day 2-4: Demo and Practical hands-on training)

METHODS & WORKFLOWS covered:

  • 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)
  • TEM/SEM screening workflows for various types of samples
  • Fine structure immuno-gold labeling (Direct immuno-EM)