Ultramicrotomy for room temperature and cryo applications
Dr. Nalan Liv
Assistant Professor of Cell Biology University Medical Center Utrecht
Jan de Weert
Product Sales Specialist BeNeLUX Advanced Workflow Specialist EMEA Leica Microsystems
Ultramicrotomy is a proven and universally accepted technique for sample preparation for TEM, SEM, AFM, etc. It permits the fine internal structures of samples to be visualized and analyzed at nanometer-scale resolution by producing ultrathin sample sections in a fast and clean manner. A key advantage of ultramicrotomy is the size and homogeneity of the electron-transparent area within the sections and the speed at which the sections are produced.
Cryoultramicrotomy allows the sectioning of vitrified biological samples, which are preserved at the atomic level and represent the actual structure at the moment of freezing. Ultrathin cryo-sections, e.g., Tokuyasu sections, provide excellent preservation of ultrastructure and antigenicity for follow-up immuno-labeling, as well as facilitating Correlative Light and Electron Microscopy (CLEM).
In this online workshop, you will discover:
- Good practice guidelines for ultrathin sectioning for TEM/SEM and ultrathin cryosectioning of vitrified and Tokuyasu frozen samples
- How to overcome some of the challenges you may face when sample sectioning
- Why pre-preparation is crucial for maximal section quality
- How to optimize grid and section manipulation
- Why and when you should use ultrathin cryosections
- An overview of the workflow for EM sample preparation for cryosectioning
- Details on the application of immunogold labeling on thawed ultrathin cryosections (Tokuyasu method) for ultrastructural localization of proteins
- How to implement high-content on-section CLEM on thawed ultrathin cryosections