Service

Sample preparation for transmission electron microscopy

  • Sample preparation for cryo-immobilization (High-pressure freezing, Plunge freezing), or chemical fixation
  • Sectioning: ultrathin, semi-thin, serial sections
  • Positive contrasting (pre- or post-embedding) and negative staining
  • Immuno-labelling/staining (pre-embedding or labeling on plastic sections)
  • Grid preparation

Sample preparation for scanning electron microscopy

  • Chemical fixation
  • ritical point drying
  • Freeze drying
  • Cryo-immobilization (High-pressure freezing, Plunge freezing)
  • Cryo, high-vacuum transfer
  • Conductive coating (Sputter and Evaporation)

 

In our EM facility there are established protocols that maximize the preservation of morphology at the EM level for each of the major model system used (cultured cells, plant and animal tissue, materials). 

 

Facility

  • Provision of EM-specified chemicals and supplies for users
  • Support and training in the methods listed in “Service” above
  • Access to the equipment of the facility
  • Training and support in ultramicrotome and electron microscope usage
  • Support and assistance in cell morphology, ultrastructure and data interpretation

 

Imaging

Transmission electron microscope Phillips CM10

This TEM can operate with different accelerating voltages (40-100 kV in 20 kV steps) and achieves the resolution up to 0.5 nm thus being suitable for a variety of biological as well as other types of samples. Microprocessor-controlled operation unit allows easy and stable usage of the microscope. High-contrast objective lens of this instrument together with bottom-mounted dedicated high-resolution EM camera TEMCam F-416 from TVIPS provide excellent images in every-day use. Original image acquisition software from TVIPS allows for an easy operation.

 

 

 

Transmission electron microscope Phillips 410

With 0.4 nm resolution at 100 kV accelerating voltage combined with 5 Mpix side-mounted CMOS camera Phillips 410 TEM is perfectly suited for routine examination of variety of samples. Additionally there is a possibility to use a bottom-mounted plate’s camera with high-resolution Imaging Plates (Ditabis).

 

 

 

 

Scanning electron microscope Hitachi S5000

High-resolution In-Lens Cold-Field Emission SEM achieves the resolution up to 0.6 nm (at accelerating voltage 30kV). Apart from SE- and BSE-detectors it is also equipped with STEM-module. The STEM-detector allows acquisition of high-resolution transmission images from not contrasted samples along with the mass determination. For the examination of biological samples at low temperature Oxford Cryoholder CT3500 can be used. Freeze-drying system together with cryo-vacuum-transfer unit were custom developed in our facility to extend the examination capabilities of frozen biological samples. The active picture acquisition and analysis system for scanning electron microscopes (DISS5; point electronic) allows simultaneous acquisition of signals from all detectors as well as on-line analysis.

 

Scanning electron microscope Hitachi S800

High-resolution scanning electron microscope equipped with Field Emission Gun achieves the resolution up to 2 nm at an accelerating voltage of 30 kV. Big specimen chamber can accommodate samples up to 5 cm in diameter and equipped with SE- and BSE-detectors. The digital scan- and image-acquisition unit DISS4 (point electronic) allows a digital control and simultaneous acquisition from both SE- and BSE-detectors.

 

Scanning electron microscope Hitachi S450

Universal scanning electron microscope with resolution of 6 nm and big 10 cm specimen chamber can provide a quick overview of a SEM sample. Due to the ease of use it also can be used for training in SEM. It is equipped with digital camera to achieve fast SE-image acquisition.

 

 

 

Scanning electron microscope CamScan CS2

Universal scanning electron microscope with accelerating voltages 0.5-40 kV achieving resolution up to 4 nm on the specimens up to 10 cm in diameter. Together with SE- and BSE-detectors our CS2 is also equipped with EDX-detection system for the microanalysis. The energy-dispersive X-ray spectroscopy allows detection of elements distribution in the specimen. The control is performed digitally through the active picture acquisition and analysis system for scanning electron microscopes (DISS4; point electronic) which permits the simultaneous acquisition of signals from several detectors.

 

Atomic force microscope JPK NanoWizard II

NanoWizard® II provides the highest resolution and stability and appeared to be the leading solution for the applications which would require the integration of modern optical methods with highest efficiency of AFM. The area of use includes mapping of living cells, experiments on single molecule or nano-particle level in biophysics, pharmacology and cellular biology. It utilizes also the patented DirectOverlay™-function which allows the direct combining of AFM- and optical microscopy images distortion free. (For more information please follow the link: http://www.jpk.com/nanowizard-r-ii-overview.351.en.html)

 

Sample preparation equipment

There is a variety of different instruments used for the sample preparation either for SEM or TEM. This includes:

  •        Fresh and fixed tissue preparation: vibrating blade microtome Leica VT1000 S
  •        Room-temperature ultramicrotomes (Reichert, Reichert Cryo, Leica Ultracut EM UC7)
  •        High-pressure-freezing machine HPM-100 with optical stimulation (Leica)
  •        Freeze-substitution machine AFS-2 (Leica)
  •        Plunge freezer
  •        Sputter coater
  •        Thermal and electron beam evaporators
  •        High vacuum cryo shuttle
  •        Critical-point dryer
  •        Freeze-drying machines