20.109(F13): Mod 3 Day 3 TEM: Difference between revisions

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=<center>TEM</center>=
=<center>TEM</center>=
==Introduction==
==Today's Goal==
The Transmission Electron Microscope (TEM) achieves its remarkable resolution by “illuminating” samples using an electron beam in a vacuum rather than using a conventional light source in air. Since the electron beam passes through the sample that is being examined, the sample must be sufficiently thin and sufficiently sturdy to be hit by electrons in a vacuum. It’s important to remember that many biological materials are damaged or destroyed by the incoming electrons and that the TEM can image only the species that survive this harsh treatment. The denser parts of the sample will absorb or scatter some of the electron beam, and it’s the scattered electrons or those that pass through the sample that are focused using an electromagnetic lens. This “electron shadow” then strikes a fluorescent screen, giving rise to an image that varies in darkness according to the sample's density. For samples that are amenable to TEM, this form of examination can allow observation of angstrom-sized objects and of cellular details down to near atomic levels.[[Image:Macintosh HD-Users-nkuldell-Desktop-TEMgrid.jpg|thumb|left|Copper TEM grid with carbon mesh, photographed at 60X magnification]]
Today we will be imaging our nanocomposites via TEM to confirm that our fabrication last week was successful.


Samples were applied last time to a wafer-thin "grid" and today they will be loaded into the TEM and placed under vacuum. The grid can be made of many kinds of materials. All have lines of a conductive metal, in our case copper, that disperse the electron beam and thereby help keep the sample from being blown to bits by the energy in the beam. A carbon mesh is strung between the metal lines. Once a sample has been applied to the grid, it's only the portions that come to rest on the carbon mesh can be visualized, along with any imperfections in the carbon mesh itself.
==Information gained via TEM==
The Transmission Electron Microscope (TEM) enables 0.14 nm resolution by utilization of an electron beam over a light source found in optical microscopes.  At this resolution, one can distinguish individual planes of atoms making TEM an excellent materials characterization instrument.  As it is possible to visualize single atoms, one can use TEM to determine a slew of parameters which describe the structure of the material, such as crystal plane spacing. Additionally, TEMs can be equipped to perform energy-dispersive X-ray spectroscopy analysis (EDX/EDS/XEDS) or electron energy-loss spectroscopy analysis (EELS).  The former, EDX, enables identification of the atomic composition of heavier elements in material. EDX has limited detection of elements below Na on the periodic table and no detection of elements below C on the periodic table. The latter, EELS, is the most sensitive to lighter elements, typically carbon to the 3d transition metals.  EELS can reveal atomic composition, electronic properties of the valence and conduction band of a material, surface properties, and chemical bonding.    


Although we will be using TEM for materials characterization, it should also be noted that TEM is used for the analysis of biological materials as well.  Since the electron beam passes through the sample that is being examined, the sample must be sufficiently thin and sufficiently sturdy to be hit by electrons in a vacuum.  Rather than imaging single atoms, biological samples display contrast of components via different densities.  Areas of higher density show up as a darker color than areas of low density, creating a “shadow ima
==TEM function==
The principle behind transmission electron microscopy is similar to the principles behind an optical microscope.  Instead of light and glass lenses, however, the TEM utilizes an electron beam focused via electromagnetic lenses.  Since the TEM utilizes an electron beam over visible light, the microscope must also function under a vacuum.  This allows the electrons to remain traveling in a straight line as they have no air particles to collide with. 
Specifically, at the top of the column of a TEM, a high voltage electron beam is emitted by a cathode and focused via condenser lens.  Then the beam is transmitted through the specimen.  This image is then magnified through a series of electromagnetic lenses until it is recorded by a fluorescent screen or camera.
==Sample preparation, using the TEM==
We will be imaging our phage nanocomposites via TEM.  To do this, we will first place samples on a TEM grid.  TEM grids can be made of many kinds of materials.  Each TEM grid design, however, includes lines of a conductive metal which disperses the electron beam and thereby prevents destruction of the sample from the harsh beam.  Our TEM grids are composed of a copper mesh with carbon mesh is strung between the grids. Once a sample has been applied to our TEM grids, we will only visualize the portions of the sample contacting the carbon mesh, along with any imperfections in the carbon mesh itself.


==Protocols==
==Protocols==

Revision as of 09:00, 6 November 2013


20.109(F13): Laboratory Fundamentals of Biological Engineering

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TEM

Today's Goal

Today we will be imaging our nanocomposites via TEM to confirm that our fabrication last week was successful.

Information gained via TEM

The Transmission Electron Microscope (TEM) enables 0.14 nm resolution by utilization of an electron beam over a light source found in optical microscopes. At this resolution, one can distinguish individual planes of atoms making TEM an excellent materials characterization instrument. As it is possible to visualize single atoms, one can use TEM to determine a slew of parameters which describe the structure of the material, such as crystal plane spacing. Additionally, TEMs can be equipped to perform energy-dispersive X-ray spectroscopy analysis (EDX/EDS/XEDS) or electron energy-loss spectroscopy analysis (EELS). The former, EDX, enables identification of the atomic composition of heavier elements in material. EDX has limited detection of elements below Na on the periodic table and no detection of elements below C on the periodic table. The latter, EELS, is the most sensitive to lighter elements, typically carbon to the 3d transition metals. EELS can reveal atomic composition, electronic properties of the valence and conduction band of a material, surface properties, and chemical bonding.

Although we will be using TEM for materials characterization, it should also be noted that TEM is used for the analysis of biological materials as well. Since the electron beam passes through the sample that is being examined, the sample must be sufficiently thin and sufficiently sturdy to be hit by electrons in a vacuum. Rather than imaging single atoms, biological samples display contrast of components via different densities. Areas of higher density show up as a darker color than areas of low density, creating a “shadow ima

TEM function

The principle behind transmission electron microscopy is similar to the principles behind an optical microscope. Instead of light and glass lenses, however, the TEM utilizes an electron beam focused via electromagnetic lenses. Since the TEM utilizes an electron beam over visible light, the microscope must also function under a vacuum. This allows the electrons to remain traveling in a straight line as they have no air particles to collide with.

Specifically, at the top of the column of a TEM, a high voltage electron beam is emitted by a cathode and focused via condenser lens. Then the beam is transmitted through the specimen. This image is then magnified through a series of electromagnetic lenses until it is recorded by a fluorescent screen or camera.

Sample preparation, using the TEM

We will be imaging our phage nanocomposites via TEM. To do this, we will first place samples on a TEM grid. TEM grids can be made of many kinds of materials. Each TEM grid design, however, includes lines of a conductive metal which disperses the electron beam and thereby prevents destruction of the sample from the harsh beam. Our TEM grids are composed of a copper mesh with carbon mesh is strung between the grids. Once a sample has been applied to our TEM grids, we will only visualize the portions of the sample contacting the carbon mesh, along with any imperfections in the carbon mesh itself.

Protocols

We're fortunate to have an expert from Angie Belcher's lab who will run the TEM grids you prepared. We've reserved the 2010FEG TEM, which is located in the basement of Bldg 24. Since the room that houses the instrument is small, we'll head over in our "supergroups," starting at about 1:30. The grids will take less than an hour to visualize and when you're not at the TEM, you should be working with your lab partner on your research proposal.
Here are some examples of the kinds of images you'll get at TEM today:

DONE!

For next time

REQUIRED ASSIGNMENT

Your team's research pre-proposal is due in lecture, M3D4. Please print out a copy of your wiki page as well as be prepared to show the page to the class and talk about it briefly. This is a great opportunity to get feedback and help with your ideas.

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