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en:aree:microscopia:elettronica:start [2018/09/05 15:26] Gianluca Frustagli |
en:aree:microscopia:elettronica:start [2020/02/06 10:32] (current) Gianluca Frustagli |
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| **Microscopy Area** | **Microscopy Area** | ||
| - | ====== Electron Microscopy ====== | + | ====== Electron Microscopy Unit ====== |
| - | ===== Instrumentation ===== | + | <WRAP tabs> |
| + | * [[start|Lab resources]] | ||
| + | * [[servizi|Servicies]] | ||
| + | * [[tecniche|Techniques]] | ||
| + | * [[richiesta|Service request]] | ||
| + | * [[chi_siamo|Contacts]] | ||
| + | </WRAP> | ||
| - | {{:aree:microscopia:elettronica:tem.jpeg?200 |}} The <wrap em>Electron Transmission Microscopy (TEM)</wrap> allows to obtain, from a sufficiently thin sample (<100nm) characterized by an appropriate electron contrast, high resolution images produced by high energy electrons (up to 100 KeV) transmitted on a fluorescent screen. | + | ==== Lab resources ==== |
| - | The TEM service provides for the use of: | + | \\ |
| - | * a **Philips EM 208S (FEI)** transmission electron microscope with tungsten source and magnification up to 200K, equipped with **MegaViewII (Olympus Soft Imaging Solutions)** camera for image acquisition; | + | <WRAP justify> |
| - | * a **LEICA UC6** ultramicrotome for sectioning samples included in epoxy and acrylic resin. | + | {{:aree:microscopia:elettronica:tem.jpeg?200 |}} <wrap em>Transmission Electron Microscopy (TEM)</wrap> allows to obtain high resolution images by high energy transmitted electrons from an ultrathin (<100nm) and adequately contrasted sample. |
| - | \\ \\ \\ \\ | + | TEM service is equipped with: |
| - | {{ :aree:microscopia:elettronica:sem.jpeg?300|}} <wrap em>Scanning Electron Microscopy (SEM)</wrap> allows to obtain three-dimensional high-resolution images from a suitable sample. The principle on which this type of microscope is based is to send a beam of primary electrons on a sample made conductive and to collect the image of the investigated surface. The image obtained is provided with a depth of field such as to allow the detailed observation of the morphological characteristics of the analyzed sample. | + | * Transmission Electron Microscope **Philips EM208S (FEI)** with a tungsten filament as electron source and x200K maximum magnification; |
| + | * image acquisition system **MegaView III** (Olympus Soft Imaging Solutions); | ||
| + | * **Ultramicrotome LEICA UC6** for sectioning of samples embedded in epoxy and acrylic resins. | ||
| - | The SEM service is equipped with the following instrumentation: | + | \\ \\ \\ \\ \\ \\ |
| - | * a high-resolution scanning electron microscope **INSPECT F (FEI)** with field emission source and magnification power up to 200K, for ultrastructural analysis of biological systems such as bacteria, parasites, cell systems, tissues; | + | <wrap em>Field Emission Scanning Electron Microscopy (FE-SEM)</wrap> allows to obtain high resolution three dimensional images from an adequately treated sample. A very high collimated electron beam interacts with a conductive sample scanning an area from which it induces the emission of secondary electrons. \\ These are collected to produce a such high focal depth image to permit detailed analysis of the morphological features of the region of interest. |
| - | * the tools needed to prepare the samples: Critical Point Drying, Sputtering, Carbon Evaporator. | + | |
| \\ | \\ | ||
| - | <WRAP box> | + | {{:aree:microscopia:elettronica:sem.jpeg?0x210|INSPECT F (FEI)}} |
| - | ==== History ==== | + | {{ :aree:microscopia:elettronica:sem-gemini.jpeg?0x210|GeminiSEM 450 (ZEISS)}} |
| - | //The electron microscopy service is offered thanks to a shared management agreement between the <wrap em>FAST</wrap> and the <wrap em>FARVA</wrap>, <wrap em>MEGE</wrap> and <wrap em>TISP</wrap> centers. In all these structures there are in fact human resources and knowledge coming from the historical area of electron microscopy. The agreement safeguards a cultural heritage that has been present in the Institute for years, allows the advancement of the electron microscopy sector and facilitates the optimization of resources.// | + | \\ |
| - | </WRAP> | + | |
| - | ===== Services ===== | + | SEM service is equipped with: |
| - | The Electron Microscopy unit offers the following services: | + | * An high resolution field emission scanning electron microscope **INSPECT F (FEI)** with a resolving power of 1.2nm in SE and 2.5nm in BSE at 30kV. This allows us to examine in addition to the surface details of biological systems such as bacteria, parasites, cells and cell growth patterns, cell-pathogens interactions, tissues; |
| + | * An high resolution field emission scanning electron microscope **GeminiSEM 450 (ZEISS)**, with Beam Booster technology and a resolving power of 0.5 nm in SE at 15 kV, enabling high resolution imaging from 0.02 a 30 kV. SEM is equipped with: InLens SE and Energy selected Backscatter detectors; VPSE and nanoVP detectors; annular STEM detector; Array Tomography (ATLAS 5). \\ The high Gemini technology allows high resolution analysis even of uncoated biological samples without charging effects. \\ The system is equipped with software for Correlative Light and Electron Microscopy (CLEM) which couples the GeminiSEM 450 and the confocal microscope LSM980. | ||
| + | * Instruments for the sample preparation: Critical Point Drying, Sputtering, Vacuum Evaporator. | ||
| - | * technical advice for the evaluation of the most suitable electron microscopy method to be used according to the proposed subject; | + | </WRAP> |
| - | * sample preparation and observation: the electron microscopy unit manages all the sample preparation steps based on the most suitable choice of the observation method; | + | |
| - | * interpretation of the results: the digital images obtained will be interpreted, in collaboration with the interested researchers, in order to obtain as much information as possible; | + | |
| - | * image processing in tables. | + | |
| - | + | ||
| - | ===== Possible applications ===== | + | |
| - | + | ||
| - | * Electron microscopy in negative contrast of freshly observable samples, such as some types of biological macromolecules, exosomes and purified cell organelles, viruses, bacteria. =>//[[.:galleria:01|gallery...]]// | + | |
| - | * Traditional transmission electron microscopy for the ultrastructural analysis of biological samples included in epoxy resin. =>//[[.:galleria:02|gallery...]]// | + | |
| - | * Scanning electron microscopy for ultrastructural analysis of the surface morphology of biological samples appropriately rendered conductive. =>//[[.:galleria:03|gallery...]]// | + | |
| - | * Immunomicroscopy on fresh preparations visualized at the TEM with negative/positive contrast, for the localization of surface antigens through the use of secondary antibodies labeled with colloidal gold particles. =>//[[.:galleria:04|gallery...]]// | + | |
| - | * Electron microscopy in pre- and post-embedding on ultra-thin sections, for the localization of both intracellular and superficial molecules, through the use of secondary antibodies labeled with colloidal gold particles and visualized at TEM. =>//[[.:galleria:05|gallery...]]// | + | |
| - | * Immunomicroscopy on whole cells appropriately treated for the localization and distribution of surface antigens through the use of secondary antibodies labeled with colloidal gold particles and detected at SEM by means of backscattered electron signals (Z contrast). =>//[[.:galleria:06|gallery...]]// | + | |
| \\ | \\ | ||
| - | === To ask for the service: === | + | <WRAP box> |
| + | ==== History ==== | ||
| - | - preliminary contact the Electron Microscopy unit staff; | + | //The electron microscopy service is offered thanks to a shared management agreement between the <wrap em>FAST</wrap> and the <wrap em>FARVA</wrap>, <wrap em>MEGE</wrap> and <wrap em>TISP</wrap> centers. In all these structures there are in fact human resources and knowledge coming from the historical area of electron microscopy. The agreement safeguards a cultural heritage that has been present in the Institute for years, allows the advancement of the electron microscopy sector and facilitates the optimization of resources.// |
| - | - fill out the online form '**[[richiesta|Service Request]]**' briefly illustrating the scientific problem, type and number of samples etc.; | + | </WRAP> |
| - | - plan a meeting with the unit staff to discuss methods, costs, timing etc. | + | |
| - | + | ||
| - | \\ | + | |
| - | + | ||
| - | > **Contacts:** | + | |
| - | > [[lucia.bertuccini@iss.it|Lucia Bertuccini]] | + | |
| - | > [[francesca.iosi@iss.it|Francesca Iosi]] | + | |
| - | > ☎ +39 06 4990 2092 | + | |
| - | > Where we are: building 1, floor C, room 67 | + | |
| - | > FIXME **To be revised**\\ //(remove this paragraph once finished)// | + | /* > FIXME **To be revised.**\\ //(remove this paragraph once finished)// */ |
