A high-speed 3D imaging microscope developed by researchers at the University of California, Santa Cruz can capture detailed cell dynamics of an entire small whole organism at once. The ability to image 3D changes in real time over a large field of view could lead to new insights in developmental biology and neuroscience.
“Traditional microscopes are constrained by how quickly they can refocus or scan through different depths, which makes it difficult to capture fast, 3D biological processes without distortion or missing information,” said Eduardo Hirata Miyasaki, who performed the work while in Sara Abrahamsson’s lab at the University of California Santa Cruz (UCSC). “Our new system extends the multifocus microscopy (MFM) technique Abrahamsson developed by using a 25-camera array to push the limits of speed and volumetric imaging. This leap in efficiency opens the door to studying small living systems in motion without disrupting them.”
The researchers describe their new microscope, which combines diffractive optics with 25 tiny cameras to synchronously and simultaneously image at multiple depths. They demonstrate live imaging of 25-plane 3D volumes measuring up to 180 × 180 × 50 μm at acquisition speeds of more than 100 volumes per second.
“The new microscope, which we call the M25, is particularly useful for imaging swimming C. elegans worms, a model organism used to study development, neuroscience and locomotion,” said Hirata Miyasaki, now at the Chan Zuckerberg Biohub. “Traditionally, scientists could only see part of the organism clearly at any one time. With our new microscope, it is possible to watch the entire worm move naturally in 3D, allowing researchers to study how its nervous system controls movement and how behavior might change in response to a genetic mutation, disease or drug treatment.”
A key part of the new microscope is the diffractive optical elements used to distribute the various focal planes across an array of 25 cameras. Diffractive optics use microstructures to manipulate light, allowing more complex light control via a thinner, lighter component than traditional optical components such as prisms.
Building upon the original MFM technique, the researchers designed a multi-focus grating to split the incoming light so that each camera captures the same scene but with a focus at a different depth. They also made customized gratings to use in front of each camera lens to correct the chromatic dispersion introduced by the multi-focus grating. By replacing the traditional chromatic-correcting prism, which was difficult to scale beyond 3×3 arrays, these blazed gratings enabled high-resolution, high-speed bioimaging across more planes.
The gratings are made from nanometer-scale patterns that require specialized fabrication tools. After using simulations to determine the optimal designs, the researchers used the University of California Santa Barbara nanofabrication facility to etch the patterns into glass. With the fabrication process now established, these diffractive elements can be accurately reproduced at higher volumes.
“One of the key innovations of the M25 is its use of simplified chromatic correction architecture: By replacing bulky prism-based components with custom-designed blazed gratings, the system achieves efficient dispersion correction across all focal planes while remaining compact and scalable,” said Abrahamsson. “This streamlined optical design not only enables high-speed imaging but also supports compatibility with label-free modalities — a major advantage for applications like embryology, where minimally invasive imaging is essential.”
“When combined, the 25 images — all acquired simultaneously, with no mechanical scanning or moving parts — form a complete 3D snapshot,” said Hirata Miyasaki. “Because this happens at high speed, limited only by the camera’s acquisition speed and the sample’s brightness, we can record entire volumes over time, enabling studies of real biological dynamics.”
The M25 microscope can be used for both fluorescence and label-free modalities, such as brightfield and polarization microscopy, which are especially useful for imaging sensitive biological systems without introducing dyes or labels. This compatibility with minimally invasive techniques makes the M25 well-suited for applications like embryology, where preserving native physiology is critical.
To validate the instrument, the researchers built a prototype and confirmed that it could capture 25 distinct, evenly spaced focal planes simultaneously, without distortion or overlap, by imaging calibration targets. They also used the microscope to image live biological specimens, including common model organisms such as C. elegans, D. melanogaster and P. marinus, demonstrating real-time 3D imaging of moving organisms without the need for scanning or motion compensation.
The system mounts to the side port of a standard commercial microscope. Aside from the diffractive optics, it requires no specialized hardware, making it more straightforward to replicate than systems that rely on custom prisms or complex light path modifications.
Next, the researchers aim to further expand the system’s scale and applications. For example, they plan to use the system’s rich imaging data to train machine learning models that can identify cell states, track dynamic behaviors and detect disease-related changes directly from images.
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“Traditional microscopes are constrained by how quickly they can refocus or scan through different depths, which makes it difficult to capture fast, 3D biological processes without distortion or missing information,” said Eduardo Hirata Miyasaki, who performed the work while in Sara Abrahamsson’s lab at the University of California Santa Cruz (UCSC). “Our new system extends the multifocus microscopy (MFM) technique Abrahamsson developed by using a 25-camera array to push the limits of speed and volumetric imaging. This leap in efficiency opens the door to studying small living systems in motion without disrupting them.”
The researchers describe their new microscope, which combines diffractive optics with 25 tiny cameras to synchronously and simultaneously image at multiple depths. They demonstrate live imaging of 25-plane 3D volumes measuring up to 180 × 180 × 50 μm at acquisition speeds of more than 100 volumes per second.
“The new microscope, which we call the M25, is particularly useful for imaging swimming C. elegans worms, a model organism used to study development, neuroscience and locomotion,” said Hirata Miyasaki, now at the Chan Zuckerberg Biohub. “Traditionally, scientists could only see part of the organism clearly at any one time. With our new microscope, it is possible to watch the entire worm move naturally in 3D, allowing researchers to study how its nervous system controls movement and how behavior might change in response to a genetic mutation, disease or drug treatment.”
A key part of the new microscope is the diffractive optical elements used to distribute the various focal planes across an array of 25 cameras. Diffractive optics use microstructures to manipulate light, allowing more complex light control via a thinner, lighter component than traditional optical components such as prisms.
Building upon the original MFM technique, the researchers designed a multi-focus grating to split the incoming light so that each camera captures the same scene but with a focus at a different depth. They also made customized gratings to use in front of each camera lens to correct the chromatic dispersion introduced by the multi-focus grating. By replacing the traditional chromatic-correcting prism, which was difficult to scale beyond 3×3 arrays, these blazed gratings enabled high-resolution, high-speed bioimaging across more planes.
The gratings are made from nanometer-scale patterns that require specialized fabrication tools. After using simulations to determine the optimal designs, the researchers used the University of California Santa Barbara nanofabrication facility to etch the patterns into glass. With the fabrication process now established, these diffractive elements can be accurately reproduced at higher volumes.
“One of the key innovations of the M25 is its use of simplified chromatic correction architecture: By replacing bulky prism-based components with custom-designed blazed gratings, the system achieves efficient dispersion correction across all focal planes while remaining compact and scalable,” said Abrahamsson. “This streamlined optical design not only enables high-speed imaging but also supports compatibility with label-free modalities — a major advantage for applications like embryology, where minimally invasive imaging is essential.”
“When combined, the 25 images — all acquired simultaneously, with no mechanical scanning or moving parts — form a complete 3D snapshot,” said Hirata Miyasaki. “Because this happens at high speed, limited only by the camera’s acquisition speed and the sample’s brightness, we can record entire volumes over time, enabling studies of real biological dynamics.”
The M25 microscope can be used for both fluorescence and label-free modalities, such as brightfield and polarization microscopy, which are especially useful for imaging sensitive biological systems without introducing dyes or labels. This compatibility with minimally invasive techniques makes the M25 well-suited for applications like embryology, where preserving native physiology is critical.
To validate the instrument, the researchers built a prototype and confirmed that it could capture 25 distinct, evenly spaced focal planes simultaneously, without distortion or overlap, by imaging calibration targets. They also used the microscope to image live biological specimens, including common model organisms such as C. elegans, D. melanogaster and P. marinus, demonstrating real-time 3D imaging of moving organisms without the need for scanning or motion compensation.
The system mounts to the side port of a standard commercial microscope. Aside from the diffractive optics, it requires no specialized hardware, making it more straightforward to replicate than systems that rely on custom prisms or complex light path modifications.
Next, the researchers aim to further expand the system’s scale and applications. For example, they plan to use the system’s rich imaging data to train machine learning models that can identify cell states, track dynamic behaviors and detect disease-related changes directly from images.
Bio Photonics Research Award
Visit: biophotonicsresearch.com
Nominate Now: https://biophotonicsresearch.com/award-nomination/?ecategory=Awards&rcategory=Awardee
#MeatAnalysis #FluorescenceTech #FoodQuality #FoodSafety #SpectroscopyInFood #MeatAuthentication #RapidDetection #FoodScience #MeatFreshness #MolecularDetection #FoodIndustryInnovation #NonDestructiveTesting #FoodMonitoring #SpectroscopyApplications #QualityControl #AdvancedSpectroscopy #MeatSpoilageDetection #FoodIntegrity #SmartFoodTesting #RealTimeAnalysis #FoodAuthenticity #FoodSafetyInnovation #SpectroscopyResearch #NextGenFoodSafety #InnovativeFoodScience,
