Electron Microscopy

About us
Electron Microscopy Laboratory

The Electron Microscopy laboratory, based in the Department of Life Sciences at Siena University, has a strong history in the high resolution microscopic analysis of cell motility complexes, invertebrate ultrastructure and many other biological subjects. Rapid freezing and cryofracturing protocols have been employed for several years as a valid complement to standard procedures for both Transmission and Scanning Electron Microscopy studies. Our researches, using principally QF-DE preparations and image analysis of metal replicas, allowed significant advances in the knowledge of axoneme structure, comparative spermatology and cell functional morphology, as documented in the literature. In a major technical upgrading cofinanced by Novartis Vaccines and UNISI, we have recently set up a laboratory for cryo-electron microscopy. This includes a Philips CM 200 transmission electron microscope, with field emission gun, wide angle compustage, cryo holder for the observation of unfixed frozen hydrated samples and a TVIPS 2Kx2K cold CCD camera with EM-Menu4: a dedicated software for automatic image collection and electron tomography. The laboratory is also equipped with a Reichert ethane-plunging device for preparation of vitrified biological samples. These new additions, coupled with our already proven abilities in the preparation and analysis of QF-DE metal replicas of axonemes and other material, establishes our laboratory as one of the most modern and productive in Italy, and one competitive with most of the world’s structural biology EM laboratories.

Services

Electron Microscopy facility provides the following services: 

  • Sample processing, embedding in plastic resin
  • Ultramicrotomy of plastic sections
  • Negative staining
  • Immuno-electron microscopy
  • Sputter coating
  • Assisting in transmission electron microscopy and digital imaging
  • Electron tomography: tilt series acquisition, image alignment and reconstruction for plastic embedded samples
  • Consultation and assistance in design of EM experiments

 

Cryo EM facility service

This facility is a "user facility" and currently does not provide cryo EM service. Any parties interested in having samples evaluated for suitability for cyro EM work should contact EM facility director, Dr. Pietro Lupetti

 

Staff

P. Lupetti (Lab. Director)

E. Paccagnini (EM protocols, imaging and image processing)

A. Gradi (EM equipments maintenance)

D. Mercati (Low temperature EM protocols)

L. Masi (EM sample processing and imaging)

Electron Microscopes
Philips CM200

Philips CM200

The Philips CM200-FEG TEM is a very versatile microscope. A point resolution of 0.19 nm is well dedicated to high-resolution TEM imaging as well as conventional morphology characterization. 

 

Main features:

  • Field Emission Gun
  • Cryo stage 
  • Acceleration voltage 200 kV
  • Spherical aberration coefficient Cs 1.35mm
  • Chromatic aberration coefficient Cc 1.2 mm
  • TVIPS CCD Camera TemCam-F224HD Slow Scan CCD Camera
  • Software TVIPS EMMENU4 and EMTool
  • Super-twin objective lens
  • Single tilt holder
  • Gatan Cryo holder with cryo work station
  • Gatan 55 cryopumping unit
FEI Tecnai G2 Spirit

FEI Tecnai G2 Spirit

High contrast, high resolution TEM for 20 –120 kV operation. Suitable for 2D & 3D imaging of thin sectioned cells, cell organelles and soft matter.  Also fitted for Electron tomography and CryoEM of hydrated samples

 

Main features:

  • Resolution: 0.34nm lattice
  • Accelerating voltage: 20 to 120kV
  • Magnification: 22x to 340,000x
  • Tile angle: +/- 80°
  • Olympus Morada side-mount 2Kx4K CCD camera
  • High level of automation: Auto-Gun and automatic tuning
  • Smart Tracking Position System for sample navigation
  • Sample observation at liquid nitrogen temperature
  • Technology for sharp imaging of thicker samples
  • I-TEM software

 

Philips CM10

Philips CM10

The Philips CM-10 TEM is a stable instrument suited to both undergraduate teaching and research. This TEM possesses a goniometer specimen stage, and is  also capable of low dose operation.  The microscope is simple to operate and generates high quality, high resolution images at conventional voltages.

 

Main features

  • Accelerating voltage: 40-100kV
  • Resolution (objective lens): 0.5 nm/5.0 ångströms (point), 0.34 nm/3.4 ångströms (line)
  • Magnification: 20- 450,000X
  • Plate film camera for image acquisition
  • Low dose, bright & dark field operation modes
Philips EM400T

Philips EM400T with EDAX DX4 Energy Dispersive Spectrometer

The Philips EM400 is a stable and  reliable high contrast analogic TEM suited for imaging of biological samples. EDAX spectrometer allows elemental analyses on thin sections and properly processed TEM specimens.

 

Main Features

  • Accelerating voltage: 40-120 Kv
  • Twin lenses
  • Goniometric stage
  • Rotary sample holder
  • EDAX DX4 Energy Dispersive Spectrometer

 

 

 

Philips XL20

Philips XL20

The Philips XL 20 is a conventional SEM with mouse-driven operation in a Windows NT environment. This system provides both secondary electron and backscattered electron imaging

Main Features

  • Resolution: 4nm
  • Magnification: max. 50,000x
  • Tungsten Filament
  • Stage movement 20 x 20 mm
  • Solid state backscattered electron imaging
  • Secondary electron imaging
Sample Processing Equipments

Freeze Fracture & Freeze Etch Systems:

Balzer’s BAF 400

Balzer’s BAF 301

Ultrarapid Freezing Devices:

MedVac Cryopress

Reichert Plunge freezer

Balzer’s FSU  Freeze Substitution Unit

Ultramicrotomes:

Reichert Ultracut

Reichert Ultracut II E

LKB Nova

SEM Processing:

Balzer’s Med 010  Mini Deposition System

Balzer’s CpD 010 Critical Point Dryer