In a groundbreaking advancement at the intersection of optical engineering and biological research, scientists have unveiled a revolutionary high-speed three-dimensional (3D) imaging microscope that stands to transform the way researchers observe dynamic biological systems. This innovative instrument, known as the M25 microscope, harnesses an array of 25 synchronized cameras combined with custom diffractive optics to capture rapid volumetric images of entire small organisms in unprecedented detail and speed. This leap in imaging technology offers a new vista for developmental biology, neuroscience, and related fields by enabling real-time visualization of complex 3D biological phenomena over a wide field of view without intrusive scanning mechanisms.
Traditional microscopy techniques face significant challenges when attempting to record rapid biological events unfolding in three dimensions. Conventional systems rely on mechanical refocusing or scanning through various depths, inherently limited by the speed of these physical movements. This constraint often results in temporal lag and spatial distortion, making it difficult to capture genuinely fast processes without losing critical information. The M25 system elegantly circumvents this barrier, employing a modified multifocus microscopy (MFM) approach extended to dozens of imaging planes simultaneously, thereby capturing an entire volumetric snapshot within milliseconds.
Central to the M25’s operation is a sophisticated optical design that integrates diffractive optical elements to distribute focal planes across a two-dimensional grid of 25 miniature cameras. Diffractive optics manipulate light through microscopic nanostructures etched onto glass surfaces, enabling precise control modes unattainable by traditional components such as prisms or lenses. These microfabricated gratings serve as multitiered light splitters, allowing each camera to focus on a distinct depth plane while maintaining perfect synchronization across the entire array. Notably, this method achieves chromatic correction through custom-designed blazed gratings strategically placed before each camera lens, eliminating the bulky and less scalable prism-based dispersion correctors used in earlier systems.
The fabrication of these intricate diffractive components demands exceptional precision, achievable only through advanced nanofabrication facilities. Researchers turned to the University of California Santa Barbara’s nanofabrication center, where state-of-the-art lithographic techniques etched nanometer-scale grating patterns into glass substrates with high fidelity. Such rigorous fabrication workflows ensure reproducibility and scalability, paving the way for broader dissemination of this technology within the academic and industrial communities.
The innovation in optical hardware is complemented by equally sophisticated software systems capable of managing the substantial data throughput generated by simultaneous acquisition from 25 cameras. The custom-designed acquisition engine coordinates precise synchronization, real-time data aggregation, and efficient storage, allowing scientists to visualize entire 3D volumes at astonishing speeds exceeding 100 volumes per second. This capability marks a paradigm shift in live-cell and organism imaging, transforming time-series volumetric data acquisition from a limiting factor into a high-resolution, high-temporal-fidelity window into biological dynamics.
Demonstrations of the microscope’s capability have been particularly compelling when applied to imaging the model organism Caenorhabditis elegans, a widely studied nematode in neurological and developmental research. Previously, scientists were restricted to observing isolated focal planes or partial organism sections, leaving critical aspects of locomotion or neural activity obscured. By concurrently imaging 25 axially spaced planes in volumetric frames, the M25 grants a holistic, distortion-free visualization of the worm’s morphology and behavior in real-time, facilitating detailed studies on how neural circuits orchestrate motion and how genetic or pharmacological modifications manifest in altered motor patterns.
Beyond its applications in C. elegans research, the M25 microscope has demonstrated versatility by imaging other model organisms such as Drosophila melanogaster and Pentatrichomonas marinus. Its compatibility with both fluorescence-based modalities and label-free imaging techniques like brightfield and polarization microscopy broadens its utility, particularly for sensitive biological samples where exogenous dyes could perturb natural physiology. This label-free imaging capability is especially critical in embryology and developmental biology, where minimally invasive observation preserves native cellular states.
The microscope’s compact optical design is ingeniously engineered to retrofit onto the side port of standard commercial microscopes, minimizing the need for specialized or custom-built mechanical components. This design decision lowers barriers to adoption by standardizing integration into existing laboratory infrastructure. Its modular nature ensures that laboratories without access to elaborate optical assembly capabilities can still benefit from ultra-high-speed multifocal imaging, thereby democratizing access to cutting-edge volumetric microscopy.
Key to the system’s optical performance is the replacement of traditional chromatic correction prisms with blazed diffractive gratings. This modification not only reduces system bulk and complexity but also enhances scalability by allowing the expansion beyond the previously limiting 3×3 camera arrays to an unprecedented 5×5 grid—in effect capturing 25 focal planes simultaneously. Such scaling is vital for probing larger sample volumes without sacrificing resolution or acquisition speed.
The researchers have made detailed open-access documentation available outlining the fabrication techniques for both the multifocus gratings and chromatic correction elements, enabling replication and customization by other laboratories worldwide. Furthermore, open-source software, including camera synchronization and image processing plugins compatible with tools like napari, is provided to ease adoption and encourage further innovation in the microscopy community.
Future directions envisioned by the research team include leveraging the rich volumetric imaging datasets for advanced computational analyses. By training machine learning algorithms on these high-dimensional time-resolved images, scientists aim to automatically detect and classify dynamic cellular states, track behavioral phenotypes, and identify subtle disease signatures directly from raw image sequences. This computational augmentation promises to accelerate biological discovery and diagnostic innovation alike.
In conclusion, the M25 microscope embodies a transformative advance in optical imaging, merging innovative diffractive optics with high-density camera arrays and streamlined software control to realize real-time 3D visualization of living biological systems at speeds and resolutions previously unattainable. Its broad applicability, modular design, and open dissemination establish it as a foundational tool for studying dynamic phenomena in developmental biology, neuroscience, and beyond.
Subject of Research: High-speed volumetric microscopy and real-time 3D imaging of live small organisms.
Article Title: High-speed 3D Imaging with 25-Camera Multifocus Microscope
Web References:
- Article DOI: 10.1364/OPTICA.563617
- Fabrication details: https://zenodo.org/records/15522415
- UCSB Nanofabrication Facility: https://nanofab.ucsb.edu/
- Optica Publishing Group: https://opg.optica.org/
References:
E. Hirata Miyasaki, A. Bajor, G. M. Pettersson, M. L. Senftleben, K. E. Fouke, T.G.W. Graham, D. D. John, J. R. Morgan, G. Haspel, S. Abrahamsson, “High-speed 3D Imaging with 25-Camera Multifocus Microscope,” Optica, vol. 12, pp. 1230-1241, 2025.
Image Credits: Eduardo Hirata Miyasaki
Keywords: Cell biology, microscopy, imaging, live cell imaging, multifocus microscopy, diffractive optics, 3D imaging, high-speed volumetric imaging, label-free microscopy, developmental biology, neuroscience
