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From Nano to Macro: How does extended microscopy break through the limitations of optical microscopy?
With the advancement of science and technology, microscopy technology has also reached a new milestone. Extended microscopy (ExM), as an innovative sample preparation tool, is playing an increasingly important role in biological research. This technique not only increases the size of the sample, but also makes visible small structures that cannot be clearly identified under conventional light microscopy.
Principle of extended microscope
The core idea of expansion microscopy is to use polymer systems to expand the sample. This means that a polymer network is first introduced into a cell or tissue sample, and then the polymer network is physically expanded through a chemical reaction to increase the size of the biological structure. Research shows that today's technology can expand specimens up to 16 times their original size, a breakthrough that makes previously unattainable resolution achievable.
The biggest advantage of this technology is that it does not require specialized high-priced microscopy equipment and the cost of the required materials is relatively low.
Historical background of extended microscopy
Extended microscopy was first proposed in 2015 by MIT researchers Fei Chen, Paul W. Tillberg and Edward Boyden. Since then, many applications have emerged, mainly focusing on the analysis of biological samples.
In 2016, researchers published papers describing solutions to the limitations of traditional labeled probes for ExM, making it possible to use this technology with conventional microscopic probes. By 2021, spatially accurate in situ sequencing technology (ExSeq) based on extended microscopy technology will also be available.
Application areas and medical potential
The applications of extended microscopy are not limited to basic biological research. In terms of disease diagnosis, extended microscopy provides imaging tools that can be used for clinical samples, which can clearly display biomolecules and minute structures within cells. This allows doctors in some cases to more accurately assess pathological conditions such as renal tubular disease, early breast tumors and differentiate between normal and cancerous tissue.
In the future, with further development of technology, extended microscopy may be able to provide nanoscale morphological observations of a variety of human organ samples.
Applications of neuroscience
In neuroscience, extended microscopy has also led to many new discoveries. Researchers can zoom in on brain circuits, making it easier to map neural connections. Extracellular biomolecules, such as proteins and nucleic acids, are firmly anchored to the polymer, allowing them to be clearly imaged with ordinary microscopes after expansion.
Advantages and limitations of extended microscopy
Compared with other microscopy technologies, extended microscopy has practical and considerable cost-effective advantages. Because it does not require high equipment investment and only requires a standard optical microscope to achieve high-resolution imaging. However, the preparation process for ExM is not easy and the integrity of each step must be ensured, otherwise the clarity of the final image may be affected.
If errors are made in certain steps, the cells may lyse or expand unevenly, compromising image quality.
In summary, extended microscopy can not only significantly improve the resolution of images, but also has a wide range of application potential, from basic research to clinical diagnosis, with far-reaching impact. However, while continuously expanding its applications, we should also think about: how will extended microscopy technology change our understanding and application of biology and medicine when it becomes more mature?
