Transmission and Scanning Electron Microscopy Services

Sample Preparation, Imaging, and Consultation

Our experienced Electron Microscopy team provides investigators with state-of-the-art technologies and expertise. We offer consultation, full service specimen preparation, and training in transmission and scanning electron microscopy, or TEM and SEM. We encourage an initial intake meeting and ongoing open communication with our staff to ensure your EM imaging meets your project goals. We also welcome all users to train with us to use the microscopes independently. 

 

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 TEM  |  SEM

Schedule With Us

To schedule TEM and SEM services with the Electron Microscopy shared resource, or to get more information about how we can work with you, contact our electron microscopy team:

Electron Microscopy Contact

Transmission Electron Microscopy (TEM)

We provide users with high-resolution, high-magnification TEM images of cellular and sub-cellular structures from fixed, embedded material. Fine external details can also be quickly viewed by TEM using negative staining techniques. Individual investigators may conduct imaging on the microscope, or our staff can provide imaging services.

Examples of specimens imaged by TEM include: 

  • C. elegans in serial section
  • Tissue punches
  • Whole cell suspension (pelleted) 
  • Embedded, sectioned bacteria  
  • Whole bacteria (negative stain) 
  • Vesicle preps (negative stain) 

 

Image (Right)

  1. A cross-section image of the ventral nerve cord of the nematode Caenorhabditis elegans.
  2. Yeast cells prepared for structural analysis by high pressure freezing followed by freeze substitution.
  3. Endocytic protein endophilin induces the formation of membrane tubules on liposomes. 
  4. Cross section of fixed and embedded Drosophila sperm. 
  5. A Bacteroides fragilis cell undergoing phage infection. 
  6. Human foreskin fibroblast (HFF) cells infected with herpes simplex virus type 1 (HSV-1). Virus capsids accumulate in the nuclear compartment before budding from the nuclear membrane as they egress out of the cell.. 

All scale bars, 500 nm.

Multiple photos
Left to Right: 1) Yan Liu, Bai Lab. 2) Sample from Daniel Barrero, Biggins lab. Image from Steve MacFarlane. 3) Sample and image from Yu Wang, Bai Lab. 4) Sample and image from Courtney Schroeder, Malik Lab. 5) Sample and image from Norma Morella, Dey Lab. 6) Sample and image from Hannah Lewis, Avgousti Lab.
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TEM Services

We offer the following TEM imaging services:

  • Imaging with our 120 kV transmission electron microscope 
  • TEM tomography for 3D structural reconstruction

We offer the following specimen processing services for TEM:

  • Conventional specimen processing and embedding 
  • High-pressure freeze and automatic freeze substitution specimen processing 
  • Microwave specimen processing  
  • Semithin (0.2 to 2 µm) and ultrathin sectioning (30 to 150 nm)
  • Serial ultrathin sectioning 
  • Negative staining  
  • Vibrating blade microtomy for fresh or fixed tissue 
  • Multiple thin film coating, rotary shadowing and glow discharge 
H. pylori
H. pylori is able to live in the acidic environment of the stomach and duodenum and can cause disease there. Image by the Salama Lab, Fred Hutch
TEM Equipment

Our ThermoFisher Talos L120C transmission electron microscope has the following features:

  • 120 kV operation 
  • 4k x 4k ThermoFisher Scientific Ceta CMOS high-resolution 16M camera 
  • Cryo and low-dose capable 
  • Magnification range: 25x to 650kx 
  • Point resolution: ≤ 0.204 

Our Leica EMUC7 ultramicrotomes are capable of ultrathin sectioning (30 to 150 nm) and semithin sectioning (0.2 to 2 µm).

Our Leica VT1200S vibratome is a fully automated vibrating blade microtome for sectioning fresh or fixed tissue with the following features:

  • Maintains cell morphology, enzyme activity and cell activity under physiological conditions
  • Includes the Vibrocheck blade deflection measurement device
  • Section thickness from 1 to 999 µm

Our Leica EMPact2 high-pressure freezer vitrifies samples up to 200 µm in thickness. Other features include the following:

  • Rapid transfer system
  • Automatic freeze substitution specimen processing with the Leica AFS2

Our Denton benchtop turbo coating system has the following capabilities:

  • Full-range, low-voltage (resistive) thermal evaporation  
  • Multiple thin film coating and rotary shadowing 
  • AC glow discharge for plasma cleaning 

Our Beckman Airfuge offers small-volume, air-driven centrifugation.

Talos L120C
Talos L120C
TEM Training

We provide one-on-one training for users interested in learning to operate the Talos L120C TEM independently. We review room temperature sample loading, focusing and image acquisition using manual acquisition TIA software from Thermo Fisher. If a more automated workflow is desired, facility staff can work with users to develop a semi-automated data collection protocol using SerialEM. Users can schedule training on our iLab site as assisted use time on the TEM. We grant users who have been cleared as independent by EM shared resource staff 24/7 access to our microscope. Users must be trained and cleared to use automation software in addition to base training on the TEM.

Steve MacFarlane helps Yan Liu embed high pressure frozen C. elegans in resin for sectioning
Steve MacFarlane helps Yan Liu embed high pressure frozen C. elegans in resin for sectioning

Scanning Electron Microscopy (SEM)

Our SEM service offers a high-performance microscope for fast characterization and 3D imaging of fine surface structures on both small and large samples. We offer training and full service imaging for internal and external SEM customers. SEM operation may be conducted by individual investigators or provided by our staff. 

Examples of specimens imaged by SEM include:

  • Whole cells in culture
  • Drosophila (adults and embryos)
  • Whole cells and bacteria deposited on coverslips
  • Artificial constructs for cell culture

 

Image (Right)

  1. Surface of a fruit fly embryo showing excess highly folded membrane that will open out to allow rapid embryo growth.
  2. Engineered thick, vascularized 3D construct in collagen hydrogel. Sample from Nicole Zeinstra, Zheng Lab (UW).
  3. BCBL-1 cells in culture. 
  4. Drosophila eye at 2200x. 
A human breast cancer cell in diatomaceous earth at 4000x magnification
Left to Right: 1) Sample and image from Mitsutoshi Nakamura, Parkhurst Lab. 2) Image from Steve MacFarlane. 3) Sample and image from the Rose Lab (SCRI). 4) Sample and image from Steve MacFarlane. All scale bars, 5 microns.
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SEM Services

We offer the following SEM imaging services:

  • Imaging with variable accelerating voltages under high- or low-vacuum conditions
  • SEM of whole cells or bulk specimens (e.g., Drosophila embryos)

We offer specimen processing for SEM, including:

  • Specimen processing and dehydration using HMDS or critical point drying  
  • Sputter coating samples
Human embryonic kidney cells
A human embryonic kidney (large cell), infected with Kaposi's sarcoma-associated herpesvirus, was induced and overlaid with Bjab cells (small cells). Image by the Rose Lab, Seattle Children's Research Institute
SEM Equipment

Our JEOL JSM 6610LV scanning electron microscope has the following features:

  • 0.3 to 30 kV operation
  • Low-vacuum operation option for nonconductive specimens
  • Large specimen chamber and automated specimen stage
  • Sensitive backscattered electron detector
  • Resolution in high-voltage mode: 3.0nm (30 kV)
  • Resolution in low-voltage mode: 4.0nm (30 kV)
  • Magnification range: 5x to 300,000x

Our other SEM equipment includes:

  • Tousimis critical point dryer with fully automated operation
  • Denton Desk IV sputter coater with variable thickness coating and a gold-palladium target
JEOL JSM 6610LV scanning electron microscope
JEOL JSM 6610LV scanning electron microscope Photo by Robert Hood / Fred Hutch News Service
SEM Training

We provide one-on-one training for users interested in learning to operate the JSM6610LV SEM independently. We review sample handling and loading, stage orientation, focusing and image acquisition using the JEOL software. Users can schedule training on our iLab site as assisted use time on the SEM. We grant users who have been cleared as independent by EM shared resource staff 24/7 access to our microscope. 

Steve MacFarlane collecting SEM images on the JSM6610LV
Steve MacFarlane collecting SEM images on the JSM6610LV Photo by Robert Hood / Fred Hutch News Service
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Questions about our TEM and SEM services or how to schedule with us?