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From cells to whole organs: How is light-sheet microscopy changing our understanding of the biological world?
In the biological world, the advancement of microscopes has always been an important tool for revealing the mysteries of life. Recently, the advent of light-sheet fluorescence microscopy (LSFM) has changed this game, providing faster imaging speeds and excellent image contrast, allowing researchers to obtain clearer views in living cells and intact organs.
Light-sheet fluorescence microscopy can capture 3D images at hundreds to thousands of frames per second, which was unimaginable with previous technologies.
Compared with traditional confocal microscopy, the detection method of light sheet microscopy is completely different. In this technology, only a thin layer of the sample (usually hundreds of nanometers to several microns thick) is vertically illuminated. This design can not only reduce light damage to the sample, but also improve the contrast of the image, allowing researchers to The reader can more keenly capture subtle changes within the sample.
Basic principles of light sheet microscopy
The basic setup of a light-sheet fluorescence microscope consists of a laser beam that is expanded and focused through a cylindrical lens to form a thin sheet of light in only one direction. This thin light sheet illuminates a specific part of the sample, and the fluorescent light emitted from the sample is collected using a standard microscope objective and projected onto an image sensor.
Due to the use of plane light illumination, the imaging speed of light sheet fluorescence microscopy can reach 100 to 1000 times that of traditional point scanning methods.
Sample loading and imaging characteristics
Although light-sheet microscopy is faster, it also requires special methods of sample setup. The sample is usually suspended under the microscope or mounted on a special support device. These measures ensure that the microscope's illumination and detection light paths are properly aligned to form clear images.
Imaging modes of light-sheet microscopy typically involve moving the sample across the image plane, and for samples larger than the image sensor, lateral movement is required. In addition, researchers can further improve image resolution through multi-view imaging.
Resolution capabilities and technological progress
The lateral resolution of light-sheet fluorescence microscopy is comparable to that of traditional fire-extinguishing fluorescence microscopy, and depends on the ability of the detection objective and the wavelength of the light used. Technological advances in recent years, such as structured illumination and super-resolution microscopy, have further enhanced the capabilities of this technology and overcome the limitations of traditional microscopy.
In addition, light-sheet microscopy combined with adaptive optics technology can obtain deeper images in thick samples, opening new doors for the application of this technology.
Wide applications of light sheet microscopy
Light-sheet fluorescence microscopy plays a vital role in areas such as developmental biology, neuroscience, and drug development. This technology not only enables observation of embryonic development over several days, but also enables precise tracking of dynamic changes in cells, which is not possible with many traditional microscopes.
The advent of light-sheet microscopy has allowed us to explore and understand life like never before.
In the future, as technology advances further, light-sheet microscopy has the potential to play an important role in a wider range of biomedical research and change our understanding of the biological world. How will this powerful microscopy imaging technology affect our understanding of life processes?
