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2024

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09

Subversion of the past! Tsinghua University Dai Qionghai/Guo Zengcai/Wu Jiamin Cooperation Latest Cell

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A comprehensive understanding of physiopathological processes requires non-invasive live three-dimensional (3D) imaging on different spatial and temporal scales.However, huge data throughput, optical non-uniformity, surface irregularities, and phototoxicity pose huge challenges, resulting in inevitable trade-offs between volume size, resolution, speed, sample health, and system complexity.

On September 13, 2024, Tsinghua University Dai Qionghai, Guo Zengcai and Wu Jiamin jointly communicated inCellOnline is titled“Long-term mesoscale imaging of 3D intercellular dynamics across a mammalian organ”The research paper,The studyIntroduced a compact real-time, ultra-large-scale, high-resolution 3D colonoscopy (RUSH3D), under the low phototoxicity of 20Hz, at 8,000 × 6,000×400 μm3The volume of the achieved 2.6×2.6×6 μm3of uniform resolution.

By integrating multiple computational imaging technologies, RUSH3D improves data throughput by a factor of 13 and reduces system size and cost by several orders of magnitude. Using these advantages, the researchers observed pre-motor neural activity and cross-day visual representation drift in the mouse cortex, the formation and progression of multiple germinal centers in the mouse inguinal lymph nodes, and a heterogeneous immune response after traumatic brain injury-All of this was performed at single-cell resolution, opening up horizons for in vivo mesoscale studies of large-scale cell-cell interactions at the organ level.

 

The magnificence of life is carefully woven by millions of cells interconnected by a complex network of signaling pathways in their own environment. Observing these cell-to-cell interactions has broadened our understanding of the mystery of mesoscale life.Examples abound, such as monitoring the interaction and migration of immune cells to decipher immune responses, or tracking the flow of information across the cortex to understand perception, cognition, and other complex behaviors. Despite differences in morphology and function, these phenomena also span three-dimensional space on a near centimeter scale in mice, weaving thousands of micron-scale cells together to compose a symphony of life. Thus, enteroscopy with centimeter-scale field of view (FOV), single-cell resolution and 3D imaging capability, as well as physiologically relevant temporal resolution and low phototoxicity, is the pivot point for in vivo fluorescence imaging.
However, the challenges are multifaceted, rooted in the physical limitations of optics and the complex living environment, resulting in the lack of universally accessible live-mediated enteroscopy. First, scale-dependent optical aberrations hinder the ability of an optical system to achieve high resolution over a large field of view. Second, the non-uniform distribution of the refractive index in the tissue produces spatially varying dynamic aberrations that degrade imaging performance and prevent accurate detection of cell location and function. Third, background contamination from light scattering in packed cells reduces contrast and fidelity. Although However, the combination of nonlinear excitation or selective illumination can reduce these pollutants, but these supplements either sacrifice the physiologically relevant time resolution of large FOV or are difficult to use in mammals. Therefore, in order to reveal the multi-scale physiological and pathological adjustments and interactions, feasible, reliable, and scalable 3D high-resolution mesoscopic design is still a challenge, both in concept and in practice.


Here, the authors introduce RUSH3D, a computational live fluorescence mesoscope with a centimeter-scale field of view, high spatial and temporal resolution, 3D resolution, low phototoxicity, and aberration robustness in complex environments, with a compact and economical framework. Through the system integration of scanning light field imaging, digital adaptive optics (DAO), multi-scale background suppression (MBR), autofocus and other technologies, RUSH3D enables spatio-temporal multi-scale imaging of the collective behavior of thousands of cells in mammalian organs under different physiological and pathological conditions, with an order of magnitude improvement in data throughput. By developing the wave-optical DAO (wDAO), spatially non-uniform system aberrations and environmental aberrations with RMS errors up to 6 wavelengths can be corrected simultaneously without reducing the data acquisition speed.Thus, RUSH3D achieved a uniform single-cell resolution of about 2.6 × 2.6 × 6 μm for a long time at 20Hz. 3The volume is 8 × 6 ×.4 μm 3, The system size and cost are reduced by two orders of magnitude.
The study demonstrates the fidelity and broad application of RUSH3D in different species, including zebrafish, jellyfish and mice. In neuroscience, full cortical 3D neural recordings at single-cell resolution were achieved in mice, and a sparse seed iterative demixing (SSID) algorithm was developed for efficient distortion-corrected neuronal signal extraction. During multisensory interaction and behavioral state transitions, different neurodynamics were observed, such as consistent preparation activity predicting the onset of movement, and the representation drift in the cortical range over 3 days was studied.In immunology, the formation of multiple germinal centers (GCs) was visualized, and the migration of T cells in multiple GCs during the immune response was determined by high-speed tracking of thousands of T cells and B cells over 10 hours. On the pathological side, reverse transendothelial migration of neutrophils after traumatic brain injury (TBI) was characterized by corrective monitoring of the cortical range from the cleared skull. In summary, RUSH3D can be used as a generally available tool for large-scale in vivo mesoscale studies of cell-cell interactions in mammalian organs.

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Wang Su's team at Southeast University found that microenvironmental glial cells regulate stem cell self-renewal and differentiation by delivering iron to neural stem cells through ferritin

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The latest research by Guo Jianping/Cheng Chao/Bo Lang of Sun Yat-sen University confirms that palmitic acid can suppress virus infection by activating innate immunity!

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