Transmission Electron Microscopy (TEM)

Process At A Glance

  • Consult with staff about experiment
  • Get fixative solution from EM
  • Fix sample and bring to lab
  • Submit form for service
  • EM does specimen preparation
  • Scientist schedules microscope time
  • Microscopy-visualization

Location: Thomas Building, DE-780
Contact phone: (206) 667-4289
Contact fax: (206) 667-1166
Contact e-mail: emsr@fredhutch.org

Transmission electron microscopy (TEM) provides information on internal structures in thin-sections (70-100nm) from your specimens. Cellular and sub-cellular structures can be viewed in great detail. TEM is also useful for viewing suspensions of microscopic particles, such as bacteria or virus using negative staining techniques. Very small amounts of sample are necessary for this technique.

TEM services provided by the EM facility include:

  • Conventional specimen fixation, dehydration & embedding
  • Specimen sectioning
  • 1 micron epoxy survey sections
  • Ultra thin sectioning (70-100nm) on ultra-microtomes (using Leica EMUC7 and UCT Ultra-microtomes), and Vibrating blade microtomy with Leica VT1200S
  • Immunoelectron microscopy
  • Cryotechniques
  • High Pressure freezing
  • Freeze substitution
  • Cryo-ultramicrotomy (Leica EM FC6 cryochamber)
  • Negative Staining (Beckman airfuge)
  • Viral & Particle Quantification
  • Platinum shadowing and carbon coating (Denton desktop vacuum evaporator)
  • Microwave techniques (Ted Pella Research Microwave System)
  • Microscopy using JEOL 1400 transmission electron microscope

Microscope operation on the transmission electron microscope (JEOL JEM 1400) may be conducted by individuals (either alone or with assistance), or may be provided by EM staff. Sign-up for microscope time or viewing time with resource staff using iLab.
 

Training

Training will be provided by the EM staff on the operation of the microscope. All self users will be required to demonstrate competency on the instrument to be considered qualified to use the instrument alone. For more in depth classes in various areas of electron microscopy please see training. Links to many EM training resources are provided in the EM Resources and Training LibGuide.

Sample preparation

Every sample is different. Please consult with the EM Staff before a project is started. You can save time and money by providing references or reprints relating to EM projects similar to the one being proposed.

The EM staff provides fixative (1/2 Karnovsky's) and will handle the specimen preparation after fixation has taken place. Samples for electron microscopy should not normally be prepared with fixatives used for light microscopy, such as formalin. See our overview of a conventional specimen protocol for TEM.
 

Obtaining your Data

Images resulting from microscopy can be saved in your fred account, on the server maintained by Center's IT department.

If you do not have a ‘fred’ account, please submit the computing account application form.

Policies

Note: specimens which come to the lab after noon on Thursday are held over in fixative or an appropriate buffer until the following Monday for processing. Investigators can expect a turn-around time of two weeks depending on workload and demands in the EM Resource.

Located in the South Lake Union neighborhood of Seattle, the facility serves Fred Hutchinson, Cancer Consortium, and all other Seattle area researchers.