Electromagnetic Lens. Pole Pieces of iron Concentrate lines of Magnetic force
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1 Electromagnetic Lens Pole Pieces of iron Concentrate lines of Magnetic force
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4 Lens Defects Since the focal length f of a lens is dependent on the strength of the lens, if follows that different wavelengths will be focused to different positions. Chromatic aberration of a lens is seen as fringes around the image due to a zone of focus.
5 Lens Defects In light optics chromatic aberration can be corrected by combining a converging lens with a diverging lens. This is known as a doublet lens
6 The simplest way to correct for chromatic aberration is to use illumination of a single wavelength! This is accomplished in an EM by having a very stable acceleration voltage. If the e velocity is stable the illumination source is monochromatic
7 Lens Defects The fact that wavelengths enter and leave the lens field at different angles results in a defect known as spherical aberration. The result is similar to that of chromatic aberration in that wavelengths are brought to different focal points
8 Spherical aberrations are worst at the periphery of a lens so again a small opening aperture that cuts off the most offensive part of the lens is the best way to reduce the effects of spherical aberration
9 Diffraction Diffraction occurs when a wavefront encounters an edge of an object. This results in the establishment of new wavefronts
10 Diffraction When this occurs at the edges of an aperture the diffracted waves tend to spread out the focus rather than concentrate them. This results in a decrease in resolution, the effect becoming more pronounced with ever smaller apertures.
11 Apertures Advantages Increase contrast by blocking scattered electrons Decrease effects of chromatic and spherical aberration by cutting off edges of a lens Disadvantages Decrease resolution due to effects of diffraction Decrease resolution by reducing half angle of illumination Decrease illumination by blocking scattered electrons
12 If a lens is not completely symmetrical objects will be focussed to different focal planes resulting in an astigmatic image
13 The result is a distorted image. This can best be prevented by having as near to perfect a lens as possible but other defects such as dirt on an aperture etc. can cause an astigmatism
14 Astigmatism in light optics is corrected by making a lens with a corresponding defect to correct for the defect in another lens In EM it is corrected using a stigmator Which is a ring of electromagnets positioned around the beam to push and pull the beam to make it more perfectly circular
15 Interazioni tra elettroni e materia
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17 Electron-specimen Interactions Primary electrons X-rays Secondary (. s.e ) Electrons Cathode Luminescence Specimen E Backscattered (. b.s.e ) Electrons Auger-electrons Absorbed Electrons Transmitted electrons
18 Two Types of Electron Microscopes Scanning Electron Microscope ( SEM ) Secondary Electrons Back-scattered Electrons ( X-rays ) Transmission Electron Microscope ( TEM ) Transmitted Electrons ( X-rays ) Primary electrons X-rays Cathode Luminescence Secondary Electrons (. s.e ) Backscattered Electrons (. b.s.e ) Auger-electrons Specimen E Absorbed Electrons Transmitted electrons
19 The size and shape of the region of primary excitation can be estimated by carrying out simulations that use Monte Carlo calculations and take into account the composition of the specimen
20 An interaction volume can also be used to predict the types of signals that will be produced and the depth from which they can escape. Monte Carlo simulations of electron trajectories are based on 1) the energy of the primary beam electron, 2) the likelihood of an interaction, 3) the change in direction and energy of the electron, 4) the mean free path of the electron and 5) a random factor for any given interaction.
21 Effects of Accelerating Voltage Z = Atomic Weight E = Energy of primary beam
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25 The angle at which the beam strikes the specimen and the distance from the surface are important factors in how much of signal escapes from the specimen.
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27 The probability of an elastic vs. an inelastic collision is based primarily on the atomic weight of the specimen.
28 Interactions of electrons with atoms Elastic scattering No energy is deposited, wavelength electron unaffected Inelastic scattering Energy is deposited, inducing damage in the sample, wavelength electron increases
29 Comparison of elastic and inelastic interactions with carbon of X-rays, neutrons and electrons Atomic cross-section for carbon in biological specimens (barns = cm 2 ) Electrons interact far stronger with matter than other elementary particles, therefore electrons can image very thin objects better than other particles Electrons deposit far less energy in a biological sample, compared to X-rays, therefore electrons are less damaging Wavelength (Å)
30 Comparing scattering of electrons & X-rays Inelastic / elastic scattering events Energy deposited per inelastic event Energy deposited relative to electrons per elastic event Scattered photons per scattered electron Electrons (200 kev) 3 10 X-rays (1.5 Å) 20 ev 8 kev Current resolution 6-8 Å < 1 Å From Henderson (1995) Quart. Rev. Biophys. 28, 171
31 Optics of diffraction and imaging detector lens object diffraction object diffraction focus image Diffraction pattern
32 Apertures Advantages Increase contrast by blocking scattered electrons Decrease effects of chromatic and spherical aberration by cutting off edges of a lens Disadvantages Decrease resolution due to effects of diffraction Decrease resolution by reducing half angle of illumination Decrease illumination by blocking scattered electrons
33 Phase contrast in the TEM Contrast can arise from constructive and destructive interference of electron waves.
34 Phase contrast in the TEM
35 Contrast in electron microscopy: bright field detector Strong scatterer lens Defining apenture
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