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Bright-field transmission electron microscopy selects the central transmitted beam; deflection into excluded electron diffraction beams can make a region dark.^[ref]

Diffraction contrast is image-intensity variation caused by spatially varying electron diffraction conditions. JEOL documents a silicon semiconductor-interconnect specimen at $200\mathrm{kV}$: its differently oriented crystals are imaged in bright-field transmission electron microscopy and same-area dark-field transmission electron microscopy, alongside an electron diffraction pattern. Selecting the marked diffracted beam makes the corresponding crystal bright after it was dark. The preparation route, thickness and exposure are not supplied. The changed objective aperture selection provides a comparison within one specimen; the readout supports orientation-dependent contrast rather than a map of surface height or chemical composition.^[ref]

This comparison is useful precisely because the material has not been replaced between the two images. It does not prove that thickness and chemical composition never influence transmission electron microscopy contrast. Instead, it supplies a concrete counterexample to assigning either quantity uniquely from brightness. A proposed extension is to repeat the first aperture setting after the second: recovery of the original contrast tests whether specimen motion or irradiation changed the comparison.

Consider an original illustrative model with exactly two outgoing scattering channels: a central beam $0$ and one diffracted beam $g$. Let $t\ge 0$ be specimen thickness and $\kappa >0$ a constant coupling with units of inverse length. Normalize the probability amplitudes $A_0,A_g$ to incident probability current, assume equal longitudinal speeds, and specifyThese ordinary differential equations define a lossless toy model, not a fit to the silicon example or a complete theory of crystal scattering. Differentiating once more gives $A_0^{\prime \prime }=-{\kappa }^2{A}_0$, soThe identity $p_0+p_g=1$ expresses conservation within these two scattering channels. Alternatively, substituting the ordinary differential equations into $d(|A_0{|}^2+|A_g{|}^2)/dt$ makes the two cross terms cancel. Here ideal bright-field transmission electron microscopy accepts fraction $p_0$ and rejects fraction $p_g$; ideal dark-field transmission electron microscopy accepts precisely the rejected channel. This complementarity requires the declared two-channel completeness and absence of loss.

Even with known $\kappa$, thickness is ambiguous: $\kappa t=\pi /4$ and $3\pi /4$ both give $(p_0,p_g)=(1/2,1/2)$. Near $t=0$, $p_g\simeq (\kappa t{)}^2$; extending that approximation through a full oscillation would erase the ambiguity incorrectly. The model isolates reversible transfer between probability amplitudes; it does not say that every thicker specimen must look darker.

Real dark-field transmission electron microscopy usually collects only a subset of the directions rejected in bright-field transmission electron microscopy. Other elastic scattering directions, inelastic scattering and detector losses prevent a general two-image conservation rule. To keep loss and background separate, extend the counting model explicitly. Let $\ell$ be the fraction absent from both recorded beam channels, $p_0+p_g=1$ describe the remaining fraction, $N$ be incident count per exposure, ${\eta }_j$ the detector efficiency and $b_j$ its mean additive background. Then the expected value of recorded count $n_j$ isFor separately acquired equal-illumination exposures with known $b_j,{\eta }_j$, the two corrected mean counts sum to $N(1-\ell )$, not necessarily $N$. Removal from these channels is not synonymous with physical absorption: an electron can leave the chosen acceptance or change kinetic energy without being stopped in the specimen. Background subtraction removes a mean offset, but its counting variance remains.

Mass–thickness contrast is image contrast caused by different amounts or strengths of electron scattering along the beam path. In bright-field transmission electron microscopy, a region that scatters more electrons outside the accepted transmitted beam generally appears darker.

A thicker part of one material and an equally thick region containing more strongly scattering atoms can therefore produce similar darkening. The image alone does not separate thickness from composition. Changing the collection aperture changes which scattered electrons are rejected, while electron diffraction in a crystal can add orientation-dependent image contrast that is not explained by mass and thickness alone.

Negative-stain electron microscopy makes a small specimen visible by surrounding it with an electron-scattering stain. Regions containing more stain scatter more electrons out of the accepted transmitted beam, so the specimen can appear relatively bright against a darker background in bright-field transmission electron microscopy.

A typical preparation deposits a dilute sample onto a supported grid, removes excess liquid, applies stain and removes excess stain before examination in a transmission electron microscope.^[ref] The resulting outline is useful for checking particle size, aggregation and gross shape. Drying, flattening and stain distribution can change the appearance, so an apparent cavity or fine boundary need not reproduce the hydrated specimen's structure.

An objective aperture selects outgoing electron directions in the objective lens's back focal plane. Passing the central transmitted beam gives bright-field transmission electron microscopy; selecting a separated diffracted beam gives dark-field transmission electron microscopy.^[ref]

In a paraxial approximation, an outgoing angle $\theta$ maps approximately to position $r=f\theta$ in this focal plane, where $f$ is the objective focal length. A centred opening of radius $a$ therefore admits $|\theta |\lesssim a/f$. Selecting a direction selects a component of the outgoing wavefunction; the subsequent image still assigns intensity to specimen positions. The opening is not a hole cut into a real-space specimen image. Its angular selection must be distinguished from selecting a specimen region for electron diffraction.

A scanning transmission electron microscope makes an image by assigning a transmitted-electron signal to each position of a scanned probe. Its essential chain is electron source, probe-forming optics, scan system, thin specimen, transmitted-electron detectors and synchronized acquisition.

An electron gun and accelerating supply provide a stable beam of the chosen kinetic energy. Condenser lens optics and an aperture control probe current and convergence; the probe-forming objective lens focuses a small spot on the specimen. Alignment, focusing and astigmatism correction keep the probe usable across the field.

Scan coils deflect the probe in a raster. A stable specimen holder positions an electron-transparent region, and a high-vacuum column limits gas scattering and contamination. A bright-field transmission electron microscopy detector collects the central transmitted beam; an annular detector can select annular dark-field electron imaging, including high-angle annular dark-field imaging. Different angular selections give different contrasts, rather than a universal map of height.

Detector electronics, scan timing and a computer assign the collected signal to the corresponding image pixel. Spectrometers are useful additions but not essential to basic image formation. The post-specimen optics can adjust detector collection geometry; they do not set scan-image magnification in the same way as the projection lenses of conventional transmission imaging.

This is a conventional transmitted-beam bright-field transmission electron microscopy image containing high-resolution electron phase contrast and visible lattice fringes, rather than an element-specific chemical map. Relative to the scale bar, the smaller particle has an outer projected diameter of roughly $40\mathrm{nm}$. It is approximately rounded and contains a denser outer rim with several lighter internal regions separated by material. A porous or internally hollow particle is a plausible morphological interpretation; the image is not a simple featureless solid sphere.

Greater projected thickness generally removes more intensity from the transmitted beam, producing darker mass–thickness contrast. Different crystallite orientations also scatter intensity differently and give diffraction contrast. Local lattice fringes reveal crystalline material and changes of fringe orientation indicate more than one coherent domain. Bright/dark edge fringes also contain defocus-dependent Fresnel contrast; their width is not a direct shell-thickness measurement. Neither an exact alumina polymorph nor three-dimensional closed porosity can be uniquely established from this projection alone. Use calibrated fringe spacings and electron diffraction for phase identification, tilt-series imaging for void geometry, and a through-focus or dose series to test contrast artefacts.

For bright-field transmission electron microscopy, place the objective aperture in the back focal plane around the transmitted beam, excluding most diffracted beams. Adjust the intermediate lens to image the objective's real-space image plane; projector lenses enlarge that image onto the screen or camera. Regions removing electrons from the admitted beam tend to be darker, although phase contrast can also contribute.

For selected-area electron diffraction, put the selected-area aperture in the objective image plane to choose a specimen region. Remove or open the objective aperture and change the intermediate-lens excitation to image the objective's back focal plane onto the camera. The projector then magnifies the diffraction pattern, rather than the specimen image. Thus the two apertures act in different conjugate planes.

Use conventional bright-field transmission electron microscopy, with nearly parallel illumination, a suitably small objective aperture and low magnification covering the whole cross-section. More strongly scattering material sends more intensity outside the selected central beam and therefore looks darker. Tilt crystalline layers away from strong Bragg diffraction conditions to reduce diffraction contrast, and use a near-focus image so that electron phase contrast does not dominate.

The nominally uniform $150\mathrm{nm}$ thickness helps separate composition from thickness variations, but does not make the contrast purely compositional: density, atomic number and multiple scattering still matter. Choose an accelerating voltage sufficient to transmit this thickness, commonly $200$–$300\mathrm{kV}$ for a robust inorganic film, subject to its damage limit. High-angle annular dark-field imaging in scanning transmission electron microscopy is a useful alternative for approximately monotonic mass-thickness contrast, but is a different acquisition mode and still requires thickness and detector-angle control.

Diffraction contrast arises because elastic scattering transfers transmitted electron intensity between the direct beam and Bragg diffraction beams. A local change in crystal orientation, strain or thickness changes that transfer. An objective aperture selecting the direct beam makes strongly diffracting regions relatively dark in bright-field transmission electron microscopy; selecting a diffracted beam reverses the selection in dark-field transmission electron microscopy. This is a change in scattering conditions, not necessarily a change in chemical composition.

Three characteristic effects are bend contours, thickness fringes and dislocation contrast. Bend contours follow positions where a gently curved crystal meets a particular Bragg diffraction condition. Thickness fringes arise from thickness-dependent exchange of intensity between coupled transmitted and diffracted waves. A dislocation distorts nearby planes, changing their diffraction condition and producing localized contrast; changing the diffraction vector can suppress particular dislocations. Moiré fringes from overlapping lattices provide another acceptable example, but their coarse spacing is a beat spacing rather than an atomic spacing.