We are excited to announce the availability of an Ultrastructure Laboratory housing a JEOL 1400 Transmission Electron Microscope. The laboratory is fully equipped for the preparation, sectioning, staining, and imaging of TEM specimens.

Our highly trained TEM Technologist has over 40 years experience and can provide you with expert guidance on planning experiments incorporating ultrastructural methods.

KEY CONSIDERATIONS for Transmission Electron Microscopy (TEM)

• Integrate TEM in Research Planning: Do not treat TEM as an afterthought in your research plans! A mistake that is often made is not considering the need or importance of ultrastructural imaging during the course of a study. Because of the nature of high resolution ultrastructural imaging, proper fixation of tissues or cells is the most important step in the process and should be considered early on.
• Consultation with TEM Staff is Key to Ensuring a Successful Outcome: Schedule an appointment with TEM staff to discuss your project's goals and scope. This consultation is free and can help prevent costly backtracking later.  It is especially important to reach out prior to the start of any experiment if you anticipate that your research plan will incorporate a TEM-based approach.
• Consider Time Consumption: Understand that TEM sample processing is labor-intensive and differs significantly from routine light-level histology. The average length of time for specimen processing, embedding, and sectioning a set of 10 samples (from one project) is approximately 60 hours. Plan your timeline accordingly. This is a good reason to first consult with the TEM Core Facility Staff--- an error or oversight on your part at the earliest step in the process can lead to spending a substantial amount of time and money to only come up with no meaningful results.
• Proper Fixation is Crucial: High-quality TEM imaging starts with proper fixation. A key issue affecting the quality of TEM imaging is poor fixation. It is important to choose the correct fixative/buffer combination and fixation method (e.g. immersion, perfusion). For TEM work, fixation has to be accomplished as rapidly as possible.
  • The TEM facility can provide protocols to properly make the fixative/buffer combination for your study or supply the necessary materials for you to make your own fixative/buffer (at a cost for materials). If you choose to make your own fixative/buffer, we ask that you purchase and use the materials/reagents listed in our protocol(s).
     

    The electron micrographs above serve to demonstrate the difference between tissues that were fixed properly (A.) and tissues that were not (B.). The tissues were fixed by immersion fixation in Karnovsky’s fixative (formaldehyde/glutaraldehyde) and processed for TEM using the same embedding protocol. The key difference between the two micrographs is the lack of fine detail.

• Avoid Artifact Introduction: Care must be taken to avoid introducing artifacts into your sample. Common artifacts include:
  • Crystals from inadequate solution preparation
  • Diatoms from contaminated water
  • Bacterial contamination in cell culture
• Orientation Matters:  Understanding the proper orientation for imaging your specimen is important, especially for tissues having a high degree of heterogeneity in structure.
• Consider Block Face Size: The routine size for a TEM block face is approximately 1 x 1 mm. Although it is possible to section a block face larger than 1 x 1 mm, keep in mind that as the size of the block face increases, the likelihood of obtaining consistent 80nm thick sections decreases.

How do you access our services?

All TEM and tissue preparation services are available on a fee-for-service basis, accessible to investigators across campus.

How do you ensure a successful project?

To ensure a successful project using the JEOL 1400 Transmission Electron Microscope (TEM), it is crucial to engage with TEM Core Facility staff early in your research planning. 

• Consultation is free and highly recommended. By taking this step early in the process, you can save time and expense by avoiding the need to re-develop a sample set.  It is particularly important that you reach out to TEM Core Facility staff prior to any cell or tissue harvest if you anticipate that your studies might need an approach incorporating TEM.