Download - Transmission Electron Microscopy
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Transmission Electron Microscopy
David Stokes2DX Workshop
University of Washington8/15-8/19/2011
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Kinds of Electron Microscopes
TEM transmissive resolution determined by optics analogous to bright field light microscope detector is film or CCD
SEM reflective, surface imaging resolution determined by spot size x-ray microanalysis or EELS possible
STEM transmissive, small spot with scan coils resolution determined by spot size analogous to confocal detector is PMT x-ray microanalysis or EELS possible
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SEM vs. TEM
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TEM sectioned cell
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TEM metal shadowed molecules
250 nm
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TEM of frozen, isolated macromolecules
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TEM of 2D membrane protein crystal
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TEM images are 2D projections
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Source of electrons
Tungsten: - outer electrons of W are made free by heating the metal. - W has excellent yield when heated just below its melting temp of 3653K. - Heat to 2600K for good yield without melting and evaporating tip (saturation point) - 100 hrs with W wire. - Make filament (cathode) at neg potential (e.g. -100 kV). - As electrons boil off the W, they are repelled by the cathode. - Potential difference relative to the anode provides the accelerating voltage - Make Weihnolt several hundred volts more negative electrons pass through the aperture in the Weihnolt cap. bias controls output of electrons and shapes field - Anode is grounded so electrons are accelerated towards it. - Electrostatic field causes crossover at the anode - this is effective source size
LaB6 - has lower work function: amount of energy necessary to free electrons. - 1000K with higher electron yield and longer life. - get small beam crossover size with high flux (10x emission of W)
Field emission - single crystal of W oriented relative its xtal lattice - no heating - series of high voltage anodes draw electrons out of lattice - electrostatic lens of anodes -> 10 nm source size - need very high vacuum -> contamination - Zirconium to reduce work function, heating to reduce contamination
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from Reimer
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Properties of Electrons
c
a
c
a
ac
c
a
eUedsedsEedsFWP )(
U=accelerating voltagecharge on electron is -1.602x10-19 Coulombs, when accelerated through 1V potential difference has kinetic energy of 1.602x10 -19 Nm (joule)define this kinetic energy as 1 eV.
With accelerating voltage of 200kV – electrons have energy of 200keV (= 3.2x10 -14 joules)compare to C-C bond of 5.8x10-19 joules (3.6 eV) or lattice binding energy of 1-2 eV
Energy of electrons is work W required to move electron from anode to cathode against force F=-eE:
Speed of electrons at 100kV: v=1.64x108m/sec more than 1/2 speed of light Have to consider relativity1MeV = 2Eo m=3m0
Wavelengths of electrons ()0.0349 Å at 120kV, 0.025 Å at 200 kV, 0.01969 Å at 300 kV
compare with wavelength of light: 2000 Åor X-rays:1.5 Å for Cu K1 Å synchrotron radiation
diffraction limit forresolution ofany optical system:D = /2
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Force on an electron
electrostatic: EqF
0
electromagnetic: BvqF
0
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Typical electron lensesTypical electron lenses
single lens (e.g. condensor)single lens (e.g. condensor) split lens (e.g. objective)split lens (e.g. objective)
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Magnetic lenses like Glass lenses Magnetic lenses like Glass lenses are governed by Newton’s lens equationare governed by Newton’s lens equation
and
http://members.shaw.ca/quadibloc/science/opt05.htm
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Focal length of Electromagnetic lens
electromagnetic lens: can change focal length by changing current
glass lens: change focus by moving specimen up and down change magnification by switching lenses
)/( 2iUKf
f = focal length K = lens const, U = voltage, i = current
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Lens Strength affects focal lengthLens Strength affects focal length
A stronger lens demagnifies the image!!!
)/( 2iUKf f = focal length K = lens const, U = voltage, i = current
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Trajectory of electrons through a homogeneous B fieldBveF
sinevBF
where is angle between v and B
Generally, path is spiral, with v|| and v components
So electrons passing through given point P will intersect again at point P'
Energy of electron conserved – no change in |v|If force is to v: electron moves in a circleIf force is || to v, path unchanged.
PP' = v||Tc = vTccos,
where Tc is time to complete circle, which is independent of
P'
P
If many angles focused in same plane, get image formed
For small angles of , cos ~1 and get image formed at P'. Larger angles of reduces the distance to P' (cos = (1 - 2/2 + . . .) and thus gives rise to spherical aberration
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Real lens is non-homogeneous field - so-called bell-shaped field
Aberrations: astigmatism, coma, spherical, field curvature, distortion (pincushion/barrel)
chromatic and Contrast Transfer Function
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TEM lens configuration
Condenser Lenses:Demagnifies sourceDefine “spot size”
Objective lens:Defines image focusHas fixed magnification (20-50x)
Intermediate lens:Controls whether Image or diffraction patternIs recorded
Projector lenses:Controls image magnificationorDiffraction “camera length”
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Condenser lenses
C1 lens produces image of sourceC2 lens demagnifies onto sample
C2 aperture reduces beam currentand makes beam more parallel
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C1 lens controls spot size
Strong C1 lens produces small probe size and weak beam
Weak C1 lens produces larger probe size and bright beam
This is known as “spot size”
C1 crossover produces image of source andMag = u/v for C2 lens
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Beam convergence
UpperObjectivelens
Increasing strength of C2 lens
~parallel convergent divergent parallel
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magnification of objective: 20-50xangle for 2.5 Å resolution at 200kv: = /d = 0.025/2.5 = 10 mrad = 0.5°
most of mag range achievedwith projection lensesMuch smaller angles andaberrations therefore lessimportant
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Back focal plane of objective lens corresponds to diff pattern
Selected area aperture is at first image plane
Intermediate lens selects either BFP or 1st image plane as its object
Diff pattern or 2nd image serves as object for projector lens
Diffraction vs. Imagingcontrolled by
Intermediate lens
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Coma
For high resolution images, you want the apparent source aligned with the real source on the optical axis and the beam to run parallel to this axis. This is coma-free alignment.
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Deflectors
electrostatic electromagnetic
Deflectors produced by transverse fields: E field (parallel plate capacitor) B field (electromagnetic lens)
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deflector coils generally come in pairs
pivot points aligned pivot points misaligned
gun tilt coils
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condensorlens
specimen
source
Apertures
condensor
aperture
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A typical scattering eventA typical scattering event
( , , ) ( , )ikr
ikzi s
er e f
r
i s
Proportionality constant for scattering:is scattering cross-section for one atom (cm2/atom)
elastic, inelastic, absorption, fission
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Electron Scattering
Elastic Scattering: Rutherford scattering
+ + + + + + + +
R
zb
F
F
2
2
221
44 R
Ze
R
qqF
electrostaticforce:
Z = atomic number,but, actually need to consider screening of electrons on nuclear charge
N.B. Elastically scattered electrons have same wavelength as incident beam and are most effective for generating phase contrast image because they interfere with the unscattered beam
42 2 2 3
0
1 1( )tot
Z RZ
a
Solving Scrodinger’s equation for the potential and making approximations for screening of nuclear charge by electron cloud:
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Inelastic Scattering
1. Oscillations of molecular bonds and phonon excitations: 20 meV – 1 eV too small to generally observe given ~1 eV spread of beam energy2. Excitation of outer electrons and valence/conduction band electrons (metals): 1-50 eV3. Ionization of inner electrons (K,L,M shells) to unoccupied shells: 10 – 100s eV
Energy transferred from incident electrons to the specimen
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Why do we hate inelastic collisions?Why do we hate inelastic collisions?
Inelastic electrons are focused at “undesirable” placesAnd therefore produce a blurred image due toChromatic aberration
Inelastic scattering produces chemical changes radiation damage
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differential scattering vs. 25 keV electrons with Ar gas
scattered outside 2.4 A aperture
inelastics within aperture
elastics within aperture
unscattered
total scattering cross-sectiondifferential cross-section
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The inelastic scattering “blurs” the low spatial frequenciesThe inelastic scattering “blurs” the low spatial frequencies
electrons X-rays
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But they can be filtered out (after they damaged the specimen)But they can be filtered out (after they damaged the specimen)
-filter +filter