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Uncovering the mystery of life: How does light sheet microscopy perform 4D observation?
Light Sheet Fluorescence Microscopy (LSFM) is a groundbreaking microscopy technology that provides a powerful imaging analysis tool for biological science research. This technology not only enables observations with medium to high optical resolution, but also achieves good optical sectioning capabilities and fast shooting speeds. Unlike traditional point scanning microscopes, the core of light sheet microscopy is to illuminate only a thin area of the sample. This feature reduces light damage when photographing living samples and is suitable for long-term observation of biological samples.
Light-sheet fluorescence microscopy can acquire images at a rate of 100 to 1,000 images per second, which is unmatched by other traditional methods.
Basic principles of light sheet microscopy
The setup of a light-sheet microscope is relatively simple and relies primarily on a laser light source. The laser is focused by a cylindrical lens to form a thin sheet of light. This light sheet is at a 90-degree angle to the viewing direction and effectively illuminates through the sample.
This technique greatly reduces background signals from samples, thereby increasing image contrast and allowing researchers to more clearly observe the structure of cells and tissues.
Technology Expansion and Application
With the development of light-sheet microscopy technology, many extension methods have been proposed. For example, bidirectional light sheet technology can reduce some common imaging artifacts and achieve better three-dimensional reconstruction effects. In addition, combining light-sheet microscopy with two-photon excitation technology can further improve its penetration into thick samples and expand its application in brain imaging experiments.
In the field of developmental biology, light sheet microscopy provides the opportunity to observe embryonic development over multiple days and to reconstruct complete developmental lineage trees.
Image Characteristics and Time Tracking
The image processing capability of light sheet microscopy enables it to obtain 3D images in a short period of time, and even achieve 4D (time) observation through time deduction. Long-term observations can last for days, and this capability is critical for studying changes in biological processes.
For example, scientists can observe the heart beats of zebrafish embryos in real time and gain insights into the dynamic changes in life activities.
Future Development Direction
Light-sheet microscopy technology continues to advance, with new imaging modes and optimization techniques emerging. Among them, the addition of adaptive optics technology provides the possibility of improving the imaging quality of thick samples. In addition, combined with ultra-high resolution microscopy technology, the application of light sheet microscopy will undoubtedly continue to expand and lead a new trend in biological research.
The development of light-sheet microscopy has opened a new chapter in the study of living samples, allowing every detail of biology to be seen at a glance.
Driven by people's curiosity about life sciences, are there more undiscovered secrets waiting for us to uncover with light sheet microscopy?
