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Electron Microscopy Sciences

EMS Academy Applications

arrow13Cryosectioning/Immunogold Workshop

Five days of hands-on training for students, researchers, and microscopists who want to learn the most up to date theory and practice in cryosectioning and immunogold labeling.

Examples of the endless possibilities when doing Immunogold Staining

Immunogold Silver Staining   Pre-embedding Immunogold Labeling
Immunogold Silver Staining of E-cadherin
on a paraffin section of human skin.
Courtesy of R. Moella, Dept. of Exp. Path.,
EUR, The Netherlands
- Mouse monoclonal anti E-cadherin
- GAM lgG Ultra-Small
- Aurion R-Gent SE-LM
  Pre-embedding Immunogold Labeling
of Huntingtin Interacting Protein 3
in Mouse Brain using Aurion
GAR Fab-US and Aurion SEEM.
Courtesy of Ms Hong Yi,
Emory University,
Atlanta, GA
Immunogold Silver Staining   Immunogold Silver Staining
Immunogold Silver Staining of alpha-amylase on Lowicryl HM20 section of rat pancreas.
- Goat-anti-Rabbit, 15nm
- Ultra-small R-Gent (Dansher Method)

Scope of Class

The objective of the course is to provide researchers with the opportunity to learn the theory and practice of the use of cryosectioning with diamond knives, and immunogold labeling. Participants will process their own samples under the expert guidance of our tutors, who are experts in cryosectioning and immunogold silver staining techniques.

During the workshop, attendees will receive theory, including but not limited to, cryosectioning, immuno detection, in situ hybridization, silver enhancement, as well as background issues. There will be time for practice as well. Attendees will be able to work with their own specimens, as well as ones we will have prepared. A full review of sectioning, incubation methods, testing of antigenicity and reactivity, complete principles of immunogold labeling, as well as preparation of conjugates for EM, as well as LM, will be covered.

Facility

The EMS Microscopy Academy
Located in Hatfield, Pennsylvania, the Academy provides electron microscopy classes, workshops and training sessions for all fields of microscopy, including materials science and biological science.

Format

Lecture, demonstration and hands-on practice, as well as round table discussion. Participants may bring their own samples to work on during lab time.

Note: Specimens need to be "ready to use", e.g., sections of chemically fixed cells on coverslips. For each combination of primary antibody and gold conjugate that you want to test during the workshop, you need at least 3, preferably 4, specimens. This enables you to test different dilutions of the primary antibody and to check the immunogold/silver reagents (negative control).

For pre-embedment immuno labeling also include specimens that will be immuno labeled for evaluation on the light microscope. These specimens are used to check if the experiment is successful. During the workshop there is no time; there no options to further process pre-embedment specimens for EM evaluation. Plastic embedment and sectioning needs to be done at your own facility.

Main Curriculum

  • Introduction in cryosectioning
  • Theory and full review of Diamond knives
  • Sectioning and practice with section pick-up, etc.
  • The properties of gold particles and their protein conjugates
  • Theory underlying immunogold labeling protocols
  • Silver enhancement of gold particles
  • Immunogold labeling on a variety of sample preparations of OLM
  • Immunogold labeling for EM:
    • Pre-embedment immunogold labeling using ultra small gold conjugates and silver enhancement
    • Post-embedment immunogold labeling on plastic and ToKuyasu cryo sections using conventional colloidal conjugates and ultra small gold conjugates
    • Manual and automated immunogold labeling
  • Pre/Post-embedment double immunogold labeling
  • Background minimization in immunogold labeling
  • Troubleshooting

Instruments Available

Hitachi TEM Inverted Light microscope Transmitted Light Microscope
Orbital Shakers Magnetic Stirrers PH Meter

Faculty

Peter van de Plas
Peter has a background in histology and immunocytochemistry and joined Aurion in 1991. During the pioneering phase of Aurion in the early nineties, he worked closely with Dr. Leunissen in founding a firm basis for Aurion. He contributed not only to the development of product applications, but also in designing the Aurion Immunogold Silver Staining workshop. He has been invited to many international microscopy conferences and workshops, and is especially experienced in providing hands-on training. Former workshop attendees and customers appreciate Peter for his technical support on, and his thorough knowledge of, the Immunogold silver staining techniques. The Aurion Immunogold workshops in Europe, Asia, Australia, and the US have all been fully attended and very well received.

Helmut Gnaegi
Helmut's background is in engineering and he is one of the founders of Diatome, the leading supplier of diamond knives and related accessories, such as ionizers and manipulators, etc. He is also one of the leading instructors for ultramicrotomy courses (Biological and Materials) around the world.

Michael Kostrna
Michael was the program director of the Electron Microscopy Technician program at Madison Area Technical College and has more than 35 years in EM technical education and research experience. He has been training EM students for 30 years and has developed curricula and lab exercises for TEM, SEM, OLM, lab safety, introductory and advanced biological EM, EM, maintenance, and x-Ray microanalysis. He has worked with companies such as SC Johnson Polymer, Dow Chemicals, Io Genetics, Virent Technologies, ABS Global, NanoOnocology, and Microscopy Inovations, and in the process gained insight to the various applications of EM.

View our schedule to see if this course is currently offered.

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