A team from the University of California, Santa Cruz has developed a new microscopy technique that breaks through the limits of fast 3D imaging. They used 25 cameras to form a high-speed microscope that can capture real-time cellular dynamics within the entire small organism at once. This technology provides unprecedented observational tools for fields such as developmental biology, neuroscience, and motor research, and will drive biomedical research towards higher dimensions and intelligence. Traditional microscopes typically rely on mechanical focusing or layer by layer scanning at different depths when acquiring 3D images, which is a slow process that cannot capture rapidly occurring biological dynamics and can easily cause image distortion or information loss. To address this issue, the team has developed a new microscope system called M25. The system is extended based on multi focus microscopy technology, utilizing 25 synchronized cameras to simultaneously record images from different focal planes, thereby achieving high-speed 3D imaging without the need for scanning. Research has shown that the new microscope can capture data from 25 focal planes at over 100 volume frame rates per second in a 3D space of up to 180 × 180 × 50 micrometers, achieving real-time imaging capabilities. The core of the system is a specially designed diffractive optical element that can divide and guide incident light to 25 cameras, each corresponding to an independent and precisely controlled focal plane. In order to overcome the problems of large volume and difficult scalability of traditional dispersion correction components, the team designed customized glitter gratings integrated in front of each camera lens, effectively correcting the dispersion effect caused by multifocal gratings. The key innovation of the M25 system lies in replacing traditional bulky prism systems with lightweight and compact custom gratings. This design makes it particularly suitable for observing embryonic development or small model organisms that move freely. During validation, the team conducted real-time 3D imaging of various living model organisms, including Caenorhabditis elegans and Drosophila melanogaster. In the past, scientists could only see partial body structures when observing the movement of nematodes, while M25 was able to track the natural trajectory of the entire nematode in 3D space. This provides a new tool for analyzing the biological nervous system, and can further explore the effects of gene mutations, disease states, or drug interventions on animal behavior. The M25 system can be directly installed on the side ports of standard commercial microscopes, without the need for additional specialized hardware except for specially designed diffractive optical elements, significantly reducing the technical threshold for promotion. The breakthrough of real-time 3D microscopy technology has brought significant changes to biomedical research. Traditional microscopes are limited by two-dimensional imaging and static observation, making it difficult to capture the dynamic full picture of life activities. In contrast, real-time 3D microscopes enable high-resolution dynamic observation of living samples. For example, in the field of cell biology, this technology enables scientists to intuitively track the migration pathways of tumor cells, providing a new perspective for the study of cancer metastasis and infection mechanisms. In the future, this technology is expected to be deeply integrated with artificial intelligence to capture a more microscopic dynamic world in real time, promoting continuous breakthroughs in biomedical research and clinical applications. (New Society)