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.
ON THIS PAGE
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:
Theo Humphreys
Director, Electron Microscopy
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)
- A cross-section image of the ventral nerve cord of the nematode Caenorhabditis elegans.
- Yeast cells prepared for structural analysis by high pressure freezing followed by freeze substitution.
- Endocytic protein endophilin induces the formation of membrane tubules on liposomes.
- Cross section of fixed and embedded Drosophila sperm.
- A Bacteroides fragilis cell undergoing phage infection.
- 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.
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
- 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
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 EM ICE high-pressure freezer vitrifies samples up to 200 µm in thickness. Other features include the following:
- Rapid transfer system
- Light and electrical stimulation options
- 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 Cressington Carbon Coater has the following capabilities:
- Voltage controlled rod source gives multiple evaporation capability.
- Automatic evaporation control gives ease of use in a busy environment.
- Thickness monitor can be fitted to give reproducible results.
Our Pelco EasiGlow glow discharger has the following capabilities:
- Short cycle times
- Programmable glow discharge settings
- Support for negative and positive modes
Our Beckman Airfuge offers small-volume, air-driven centrifugation.
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.
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)
- Surface of a fruit fly embryo showing excess highly folded membrane that will open out to allow rapid embryo growth.
- Engineered thick, vascularized 3D construct in collagen hydrogel. Sample from Nicole Zeinstra, Zheng Lab (UW).
- BCBL-1 cells in culture.
- Drosophila eye at 2200x.
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
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
- 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
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.