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Updated: Sep 28, 2025

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography
Published on: March 12, 2017
High-Energy Electron Scattering in Thick Samples Evaluated by Bright-Field Transmission Electron Microscopy,
Misa Hayashida1, Marek Malac1,2
1Nanotechnology Research Centre, National Research Council, Edmonton, ABT6G 2M9, Canada.
Abstract:
Energy-filtering transmission electron microscopy (TEM) and bright-field TEM can be used to extract local sample thickness $t$ and to generate two-dimensional sample thickness maps. Electron tomography can be used to accurately verify the local $t$. The relations of log-ratio of zero-loss filtered energy-filtering TEM beam intensity ($I_{{\rm ZLP}}$) and unfiltered beam intensity ($I_{\rm u}$) versus sample thickness $t$ were measured for five values of collection angle in a microscope equipped with an energy filter. Furthermore, log-ratio of the incident (primary) beam intensity ($I_{\rm p}$) and the transmitted beam $I_{{\rm tr}}$ versus $t$ in bright-field TEM was measured utilizing a camera before the energy filter. The measurements were performed on a multilayer sample containing eight materials and thickness $t$ up to 800 nm. Local thickness $t$ was verified by electron tomography. The following results are reported:• The maximum thickness $t_{{\rm max}}$ yielding a linear relation of log-ratio, $\ln ( {I_{\rm u}}/{I_{{\rm ZLP}}})$ and $\ln ( {I_{\rm p}}/{I_{{\rm tr}}} )$, versus $t$.• Inelastic mean free path ($\lambda _{{\rm in}}$) for five values of collection angle.• Total mean free path ($\lambda _{{\rm total}}$) of electrons excluded by an angle-limiting aperture.• $\lambda _{{\rm in}}$ and $\lambda _{{\rm total}}$ are evaluated for the eight materials with atomic number from $\approx$10 to 79.The results can be utilized as a guide for upper limit of $t$ evaluation in energy-filtering TEM and bright-field TEM and for optimizing electron tomography experiments.
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