Transmission Electron Microscopy (TEM)- Bioimaging
Transmission Electron Microscopy (TEM) has emerged as a powerful tool in the field of bioimaging, revolutionizing our understanding of the intricate world of biological structures at the nanoscale level. Unlike conventional light microscopy, TEM utilizes a beam of electrons instead of light to visualize specimens, enabling researchers to probe into the inner workings of cells and tissues with unprecedented detail.
At the heart of a TEM instrument lies an electron gun, which emits a beam of high-energy electrons. This electron beam passes through a series of electromagnetic lenses, which focus and shape it into a narrow, coherent stream. The beam is then directed onto a thin sample, typically less than 100 nanometers thick, mounted on a grid. As the electrons interact with the specimen, they undergo various interactions, generating valuable information about its composition, structure, and morphology.
Figure 1. Transmission electron microscope (TEM) and scanning electron microscope (SEM) images of the synthetic hematite samples. (Andrew Roberts, et al.; 2021)
One of the primary interactions exploited in TEM is elastic scattering, known as the "image-forming" process. When electrons encounter atoms within the sample, they scatter in different directions, depending on the atomic arrangement and density of the material. By collecting and magnifying these scattered electrons, a detailed image of the sample's internal structure is formed. This image can reveal crucial features such as cell organelles, protein complexes, and even individual atoms.
In addition to elastic scattering, TEM also capitalizes on inelastic scattering phenomena. Inelastic scattering occurs when the electrons lose energy while interacting with the sample. This loss of energy can provide valuable spectroscopic information about the chemical composition of the specimen. By analyzing the energy loss spectrum, researchers can identify the presence of specific elements and study their distribution within the sample.
To enhance contrast and reveal fine details, various sample preparation techniques are employed in TEM. One commonly used method is staining, where heavy metal compounds are applied to the sample. These stains selectively bind to certain cellular components, creating contrast by increasing the electron scattering. Another technique is negative staining, where a contrasting material is deposited around the sample, highlighting its shape and structure. Cryo-electron microscopy (cryo-EM) is a specialized technique that preserves biological specimens in a frozen, hydrated state, avoiding the need for chemical fixation and staining. This approach has enabled imaging of delicate biological structures in their near-native conditions.
TEM has not only revolutionized the study of cellular and molecular biology but has also played a crucial role in various interdisciplinary fields. In materials science, TEM is used to examine the atomic structure and defects in materials, aiding the development of advanced materials with tailored properties. In nanotechnology, TEM is indispensable for characterizing and manipulating nanoparticles and nanomaterials, enabling precise control over their synthesis and assembly.
Despite its immense capabilities, TEM also poses several challenges. Sample preparation for TEM requires skilled techniques to obtain ultra-thin sections without introducing artifacts. The high vacuum environment inside the microscope restricts the imaging of living samples, necessitating meticulous sample fixation procedures. Additionally, the high energy of the electron beam can damage sensitive biological samples, limiting the observation time and resolution.
In conclusion, Transmission Electron Microscopy has revolutionized bioimaging by enabling the visualization of biological structures at the nanoscale. By harnessing the interactions of electrons with specimens, TEM provides unprecedented insights into the intricate world of cells and tissues. With further advancements in sample preparation techniques and instrument design, TEM continues to push the boundaries of our understanding in biology, materials science, and nanotechnology.
- Andrew Roberts, et al.; Magnetic Domain State and Anisotropy in Hematite (α-Fe2O3) From First-Order Reversal Curve Diagrams. Journal of Geophysical Research: Solid Earth. 2021, 126(12).
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