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Service Centre

Electron Microscopy

Instituto Gulbenkian de Ciência

Service Centre category(ies)

Microscopy

Description of the Service Centre

The Electron Microscopy Facility is part of the shared facilities at the Instituto Gulbenkian de Ciência, Oeiras, Portugal. The facility provides centralized, high-quality room temperature transmission electron microscopy techniques and resources for all researchers at the Instituto Gulbenkian de Ciência and within the Portuguese Research Community. Capabilities include conventional and cryo sample preservation, room temperature and cryo-sectioning and room temperature TEM.

Service Centre web address

Instituto Gulbenkian de Ciência, Rua da Quinta Grande, 6, 2780-156 Oeiras, Portugal

Contact person's name

Dra Erin Tranfield

Contact person's e-mail

etranfield@igc.gulbenkian.pt

Contact person's telephone

+351 21 446 4691

Last edited on

2015-05-08 14:54:55

Services

Service

Transmission Electron Microscopy Sample Processing and Imaging

Available for external use?

Yes

Comments

We provide full or partial service for sample processing using conventional sample processing protocols. Protocols are selected based upon the sample and the research questions and will vary between projects.

Service

Training in Transmission Electron Microscopy Techniques

Available for external use?

Yes

Comments

We offer training for new users on all aspects of electron microscopy, from sample processing, to ultra-thin sectioning, to imaging.

Equipments

Equipment

Leica UC7 / FC7 Ultramicrotome

Service type: self-service or full-service?

Full-service

Comments

The Leica UC7 / FC7 Ultramicrotome was installed in the EM Facility in February 2015. The instrument is in an isolated room, free from air currents and constant interruptions. This instrument is used for sample trimming, and cutting of ultra-thin sections (50-90 nm), and semi-thin sections (300 – 700 nm) at both room and cryo temperatures. A section thickness of 60 – 120 nm is optimal for TEM imaging on the Hitachi H-7650 located in the electron microscopy facility. The ultramicrotome room is equipped with an oxygen sensor as a safety measure for handling liquid nitrogen during cryo-sectioning. The instrument is equipped with a cryo-sphere and a micromanipulator system to improve the quality and easy of cryo-sectioning. Cryo-sectioning has priority on this instrument, however it is also available for use on room temperature samples.

Equipment

Q150T ES Carbon Coater

Service type: self-service or full-service?

Full-service

Comments

The Q150T ES Carbon Coater provides three functions in just one physical machine: • Carbon/Metal: this feature provides the possibility of creating carbon films by carbon filaments. This is convenient for Electron Microscopy due to films’ electrical conductivity and mechanical stability. Carbon coating is used frequently to sections under the electron beam thereby reducing section damage and image drift. Carbon coating is also used to prepare grids for negative staining and to prepare landmarks on surfaces used for correlative light and electron microscopy; • Glow Discharge: with this option you can turn hydrophobic surfaces (created during carbon coating) to hydrophilic surfaces. This is used frequently in the preparation of samples for correlative light and electron microscopy as well as for grid preparation for negative staining; • Sputter Coating: allows you to create a thin film, electrically conductive, on the surface of your sample. This is useful when you’re preparing your specimens for Scanning Electron Microscopy (SEM). The Carbon Coater needs special training that can be provided by one of the technicians in the IGC Electron Microscopy Facility so that users are able to mount the interchangeable inserts.

Equipment

PELCO Biowave Sample Processing System with SteadyTemp Pro

Service type: self-service or full-service?

Full-service

Comments

PELCO Biowave is a microwave processing system that allows the complete control of power delivered to the specimen. The unit has built-in extraction, vacuum and a water circulation system. It is controlled by a touchscreen with the most common protocols saved within the memory of the microwave. The SteadyTemp Pro captures and removes the heat ensuring that the samples are not damaged by increasing temperatures. Microwave processing greatly increases the speed of fixative penetration into the sample thereby reducing the time needed for conventional electron microscopy processing from several days to several hours.

Equipment

High Pressure Freezing Machine

Service type: self-service or full-service?

Full-service

Comments

The Wohlwend HPF Compact 02 allows freezing cell cultures, tissue specimens and emulsions for Transmission Electron Microscopy or Scanning Electron Microscopy. With this equipment is it possible to cryo-immobilize the sample in vitreous ice to preserve the sample in a few miliseconds. This helps to reduce alterations to sample ultrastructure, and improving the fidelity of sample preservation. At the Instituto Gulbenkian de Ciência Electron Microscopy Facility there is currently no way to perform cryo-electron microscopy thus the Wohlwend HPF is best used in combination with the Leica EM AFS2 to prepare fixed, resin embedded samples for correlative light and electron microscopy, immunogold labeling, electron tomography or conventional electron microscopy.

Equipment

Hitachi H-7650 TEM

Service type: self-service or full-service?

Full-service

Comments

The Hitachi H-7650 is a 120kv Transmission Electron Microscope (TEM) that was installed at the Instituto Gulbenkian de Ciência in 2010. The Hitachi H-7650 was especially designed for biological TEM processed specimens and is capable of magnifications as high as 600,000x and is equipped with an AMT digital camera for data acquisition. It has a dedicated room, with appropriate conditions, like stabilized temperature. This TEM provides essential features like auto-focus and auto-tilt (+/-60°) that allow you to observe 3D structures of your sample by TEM tomography. Being such a sensitive piece of equipment, the TEM needs special training that can be provided by one of the technicians in the IGC Electron Microscopy Facility.

Equipment

Automatic Freeze Substitution Unit

Service type: self-service or full-service?

Full-service

Comments

The Leica EM AFS2 is dedicated to freeze substitution and low temperature embedding and is suitable for light and electron microscopy. The Leica EM AFS2 is located in a dedicated floor mounted fumehood close to the High Pressure Freezer. With features like stereomicroscope, fume exhaust system, fast LED UV polymerization, mouse-controlled colour screen, mobile with clamping wheels and intuitive programming with USB port it is possible to freeze substitution your high pressure frozen specimens and to embed them. Freeze substitution protocols for correlative light and electron microscopy and immunogold labeling are being implemented into the electron microscopy facility.

Equipment

Various support equipment

Service type: self-service or full-service?

Full-service

Comments

The facility has many small pieces of support equipment that are used during the proce

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