Dynamic Visualization Research Articles

Dynamic visualization of DNA methylation in cell cycle genes during iPSC cardiac differentiation.

Existing analyses with conventional assays have generated significant insights into static states of DNA methylation but were unable to visualize the dynamics of epigenetic regulation. We utilized a genomic DNA methylation reporter (GMR) system carrying Snrpn minimal promoter and CpG regions of Cdk1 (Cyclin-dependent kinase 1) or Sox2 (SRY-Box Transcription Factor 2). Mouse Sox2 GMR iPSCs rapidly lost fluorescent reporter signal upon the induction of cardiac differentiation. Cdk1 GMR reporter signal was strong in undifferentiated iPSCs, and gradually decreased during cardiomyocyte differentiation. RT-qPCR and pyrosequencing demonstrated that the reduction of Sox2 and Cdk1 was regulated by hypermethylation of their promoters' CpG regions during cardiac differentiation. The GMR reporter system can be useful for monitoring real-time epigenetic DNA modification at single-cell resolution.

Artificial Intelligence-Empowered Spectroscopic Single Molecule Localization Microscopy.

Spectroscopic single-molecule localization microscopy (SMLM) has revolutionized the visualization and analysis of molecular structures and dynamics at the nanoscale level. The technique of combining high spatial resolution of SMLM with spectral information, enables multicolor super-resolution imaging and provides insights into the local chemical environment of individual molecules. However, spectroscopic SMLM faces significant challenges, including limited spectral resolution and compromised localization precision because of signal splitting and the difficulties in analyzing complex, multidimensional datasets, that limit its application in studying intricate biological systems and materials. The recent integration of artificial intelligence (AI) with spectroscopic SMLM has emerged as a powerful approach for addressing these challenges. Here, it is reviewed how AI-based methods applied to spectroscopic SMLM enhance and expand the capabilities of these applications. Recent advancements in AI-driven data analysis for spectroscopic SMLM, including improved spectral classification, localization precision, and extraction of rich spectral information from unmodified point-spread functions are discussed, further examining their applications in biological studies, materials science, and single-molecule reaction analysis, which highlight how AI provides new insights into molecular behavior and interactions. The AI-empowered approach adds new dimensions of information and provides new opportunities and insights into the nanoscale world of rapidly evolving field of spectroscopic SMLM.

Augmenting Human Expertise in Weighted Ensemble Simulations through Deep Learning-Based Information Bottleneck.

The weighted ensemble (WE) method stands out as a widely used segment-based sampling technique renowned for its rigorous treatment of kinetics. The WE framework typically involves initially mapping the configuration space onto a low-dimensional collective variable (CV) space and then partitioning it into bins. The efficacy of WE simulations heavily depends on the selection of CVs and binning schemes. The recently proposed state predictive information bottleneck (SPIB) method has emerged as a promising tool for automatically constructing CVs from data and guiding enhanced sampling through an iterative manner. In this work, we advance this data-driven pipeline by incorporating prior expert knowledge. Our hybrid approach combines SPIB-learned CVs to enhance sampling in explored regions with expert-based CVs to guide exploration in regions of interest, synergizing the strengths of both methods. Through benchmarking on alanine dipeptide and chignolin systems, we demonstrate that our hybrid approach effectively guides WE simulations to sample states of interest and reduces run-to-run variances. Moreover, our integration of the SPIB model also enhances the analysis and interpretation of WE simulation data by effectively identifying metastable states and pathways and offering direct visualization of dynamics.

Design, manufacturing, and multi-modal imaging of stereolithography 3D printed flexible intracranial aneurysm phantoms.

Physical vascular phantoms are instrumental in studying intracranial aneurysms and testing relevant imaging tools and training systems to provide improved clinical care. Current vascular phantom production methods have major limitations in capturing the biophysical and morphological characteristics of intracranial aneurysms with good fidelity and multi-modal imaging capacity. With stereolithography (SLA) 3D printing technology becoming more accessible, newer flexible and transparent printing materials with higher precision controls open the door for improving the efficiency and quality of producing anthropomorphic vascular phantoms but have rarely been explored for the application. This technical note intends to report the feasibility of using SLA 3D printing technology to manufacture flexible intracranial aneurysm phantoms with similar scales to the real anatomy, as well as their capacity for multi-modal flow imaging and analysis, including ultrasound flow imaging, high-speed filming, and particle image velocimetry analysis. We designed and 3D-printed two intracranial aneurysm phantoms with an SLA 3D printer using Formlabs Elastic 50A resin. By using a micropump to introduce cyclical flows in the phantoms, we first employed conventional Doppler and vector flow ultrasonography to observe and measure different fluidic properties. Then, a high-speed camera was used to record particles flowing within the phantom, and we further conducted a particle image velocimetry analysis, including the distribution of mean 2D velocity vectors, average velocity magnitudes, and the mean vorticity fields in the phantom for the high-speed imaging data. We successfully 3D-printed flexible intracranial aneurysm phantoms with similar dimensions to the real anatomy. Additionally, we validated the phantoms' ability to allow visualization, measurement, and analysis of flow dynamics based on both real-time ultrasound and optical imaging. Our proof-of-concept study illustrates that SLA 3D printing using commercial elastic resins can significantly contribute towards facilitating the fabrication of flexible intracranial aneurysms phantoms for training, research, and preoperative planning.

Direct visualization of the charge transfer state dynamics in dilute-donor organic photovoltaic blends

The interconversion dynamics between charge transfer state charges (CTCs) and separated charges (SCs) is still an unresolved issue in the field of organic photovoltaics. Here, a transient absorption spectroscopy (TAS) study of a thermally evaporated small-molecule:fullerene system (α6T:C60) in different morphologies (dilute intermixed and phase separated) is presented. Spectral decomposition reveals two charge species with distinct absorption characteristics and different dynamics. Using time-dependent density functional theory, these species are identified as CTCs and SCs, where the spectral differences arise from broken symmetry in the charge transfer state that turns forbidden transitions into allowed ones. Based on this assignment, a kinetic model is formulated allowing the characterization of the charge generation, separation, and recombination mechanisms. We find that SCs are either formed directly from excitons within a few picoseconds or more slowly (~30–80 ps) from reversible splitting of CTCs. These findings constitute the first unambiguous observation of spectrally resolved CTCs and SCs.

A Method for Rapid Inducible RNA Decay.

Modulating RNA decay is apowerful tool to investigate RNA degradation dynamics. Here, we describe a protocol to inducibly recruit protein factors to regulate target RNA metabolism, called Rapid Inducible Decay of RNA (RIDR). RIDR induces fast and synchronous decay of target mRNAs within minutesandenables direct visualization of mRNA decay dynamics and subcellular kinetics in living cells. Here, we provide detailed procedures to make stable cell lines, conduct fixed- and live-cell measurements, and perform data analysis. We discuss the potential pitfalls and make RIDR applicable to a general biology lab.

Exploring Biomineralization Processes Using In Situ Liquid Transmission Electron Microscopy: A Review.

Liquid transmission electron microscopy (TEM) is a newly established technique broadly used to study reactions in situ. Since its emergence, complex and multifaceted biomineralization processes have been revealed with real-time resolution, where classical and non-classical mineralization pathways have been dynamically observed primarily for Ca and Fe-based mineral systems in situ. For years, classical crystallization pathways have dominated theories on biomineralization progression despite observations of non-traditional routes involving precursor phases using traditional- and cryo-TEM. The new dynamic lens provided by liquid TEM is a key correlate to techniques limited to time-stamped, static observations - helping shift paradigms in biomineralization toward non-classical theories with dynamic mechanistic visualization. Liquid TEM provides new insights into fundamental biomineralization processes and essential physiological and pathological processes for a wide range of organisms. This review critically reviews a summary of recent in situ liquid TEM research related to the biomineralization field. Key liquid TEM preparation and imaging parameters are provided as a foundation for researchers while technical challenges are discussed. In future, the expansion of liquid TEM research in the biomineralization field will lead to transformative discoveries, providing complementary dynamic insights into biological systems.

CSF flow measurement in the mesencephalic aqueduct using 2D cine phase-contrast MRI in dogs with communicating internal hydrocephalus, ventriculomegaly, and physiologic ventricular spaces.

Brachycephalic dogs are overrepresented with ventricular enlargement. Cerebrospinal fluid (CSF) flow dynamics are not completely understood. MRI techniques have been used for the visualization of CSF dynamics including phase-contrast imaging. This study aimed to determine a causality between CSF flow and ventriculomegaly or hydrocephalus and to compare CSF flow dynamics among dogs with ventriculomegaly, internal hydrocephalus, and physiologic ventricles. A total of 51 client-owned dogs were included in the study. Magnetic resonance imaging (MRI)-based FLASH sequences and phase-contrast images of the brain were obtained, and the ROI was placed at the level of the mesencephalic aqueduct. ECG monitoring was performed parallel to MRI acquisition. Evaluation of flow diagrams and processing of phase-contrast images were performed using commercially available software (Argus VA80A, Siemens AG Healthcare Sector, Erlangen, Germany). Dogs were divided into three groups: Group 1 consisted of brachycephalic dogs with ventriculomegaly (group 1A) or internal hydrocephalus (group 1B), group 2 consisted of brachycephalic dogs with normal ventricles, and group 3 consisted of meso- to dolichocephalic dogs with normal ventricles. Group 1 had a higher median Vrost (4.32 cm/s; CI: 2.94-6.33 cm/s) and Vcaud (-6.1 cm/s, CI: 3.99-9.33 cm/s) than group 2 (Vrost: 1.99 cm/s; CI 1.43-2.78 cm/s; Vcaud: 2.91 cm/s, CI: 2.01-4.21 cm/s; p = 0.008; p = 0.03) and group 3 (Vrost:1.85 cm/s, CI: 1.31-2.60 cm/s; Vcaud - 2.46 cm/s, CI 1.68-3.58 cm/s; p = 0.01; p = 0.02). The median Volcaud of group 1 (-0.23 mL/min, CI: 0.13-0.42 mL/min) was higher than that of group 2 (-0.09 mL/min, CI: 0.05 mL/min and 0.15 mL/min) (p = 0.03). Groups 1A and 1B did not differ in Vcaud, Vrost, Volcaud, and Volrost. Group 1A and 1B showed a higher median Vrost (4.01 cm/s, CI: 2.30-7.05 cm/s; 5.94 cm/s, CI: 2.16-7.88 cm/s) than group 2 (1.85 cm/s, CI: 1.24-2.80 cm/s.) (p = 0.03; p = 0.004). Increased CSF flow velocities in rostral and caudal directions are present in dogs with ventriculomegaly and internal hydrocephalus compared to normal controls.

Version [formula omitted] — [CACP: Classification Algorithms Comparison Pipeline]

We present the first major release of the Classification Algorithms Comparison Pipeline (CACP). The proposed software enables one to compare newly developed classification algorithms in Python with other classifiers to evaluate classification performance and ensure both outcomes’ reproducibility and statistical reliability. CACP simplifies and accelerates the entire classifier evaluation process considerably and helps prepare the professional documentation of the experiments conducted. The upgrade introduces enhancements to existing tools and adds new features: (1) - support for River machine learning library datasets in incremental learning, (2) - capability to include user-defined datasets, (3) - use of River classifiers for incremental learning, (4) - use of River metrics for incremental learning, (5) - flexibility to create user-defined metrics, (6) - record-by-record testing for incremental learning, (7) - enhanced summary of incremental testing results with dynamic visualization of the learning process, (8) - Graphical User Interface (GUI).

Dynamic visualization study of in situ cleaning of polystyrene microparticles off silicon wafer surface

With the improvement of chip performance, the requirements for cleaning the surface of silicon wafers are becoming higher. However, due to equipment and technology, it is difficult to observe the complex motion processes of particles at the microscopic scale. In this paper, an in situ dynamic visualization experiment on the cleaning of Polystyrene Latex (PSL) on the surface of silicon wafers is carried out by using a high-speed camera and image processing software. The mechanical behavior of PSL particles in fluid was investigated on a microscopic scale, and the trajectory and force of the polystyrene particles on the surface of the wafers were visualized, which provided a new perspective for understanding the complex cleaning process. Theoretical models were developed to explain the motion characteristics of the particles by calculating parameters such as van der Waals force, surface tension, and trailing force, and these models provide a theoretical basis for optimizing the cleaning process. There are four particle motion modes in the fluid: (1) interface capture, where the particles on the surface of silicon wafer are trapped by gas–liquid interface under surface tension; (2) particle collision, where the particles captured by the water film collide with the particles on the wafer surface to make the latter leave the silicon wafer; (3) jump attachment, where the particles jump and attach to the surface of the particle group under the action of lift; and (4) wall surface movement, where the particles start up under the action of water flow and then leave the silicon wafer quickly.

Co-Designing Dynamic Mixed Reality Drill Positioning Widgets: A Collaborative Approach with Dentists in a Realistic Setup.

Mixed Reality (MR) is proven in the literature to support precise spatial dental drill positioning by superimposing 3D widgets. Despite this, the related knowledge about widget's visual design and interactive user feedback is still limited. Therefore, this study is contributed to by co-designed MR drill tool positioning widgets with two expert dentists and three MR experts. The results of co-design are two static widgets (SWs): a simple entry point, a target axis, and two dynamic widgets (DWs), variants of dynamic error visualization with and without a target axis (DWTA and DWEP). We evaluated the co-designed widgets in a virtual reality simulation supported by a realistic setup with a tracked phantom patient, a virtual magnifying loupe, and a dentist's foot pedal. The user study involved 35 dentists with various backgrounds and years of experience. The findings demonstrated significant results; DWs outperform SWs in positional and rotational precision, especially with younger generations and subjects with gaming experiences. The user preference remains for DWs (19) instead of SWs (16). However, findings indicated that the precision positively correlates with the time trade-off. The post-experience questionnaire (NASA-TLX) showed that DWs increase mental and physical demand, effort, and frustration more than SWs. Comparisons between DWEP and DWTA show that the DW's complexity level influences time, physical and mental demands. The DWs are extensible to diverse medical and industrial scenarios that demand precision.

QURANIC IMAGERY OF SABR (PATIENCE)

This study investigates the Quranic representation of both kinetic and abstract imagery to elucidate the significance and essence of sabr as conveyed in the verses of the Quran. Both abstract and kinetic pictures are analyzed to enhance the imagery elaboration of the abstract idea of sabr as presented in the verses of the Quran. The abstract image underwent a first evaluation to ascertain the value and quality of sabr. The study examines diverse methods of practicing sabr through abstract concepts, including confidence, hope, serenity, piety, and obedience, which are classified as faith and piety. The study identified the representation of dynamic visuals in the portrayal of the tangible concept of sabr. The concrete images of sabr are interpreted through two techniques of kinetic imagery, represented by second and third-person pronouns. The second pronoun is employed to engage the reader in vivid imagery, while the third pronoun aims to facilitate readers visualization of the narratives of sabr. This study assessed the kinetic and abstract representations in interpreting abstract qualities and values. Examining the representations of abstract concepts from the Quran is recommended to attain a more profound comprehension of them.

Dynamic synthetic-scanning photoacoustic tracking monitors hepatic and renal clearance pathway of exogeneous probes in vivo

Advancements in precision medicine necessitate understanding drug clearance pathways, especially in organs like the liver and kidneys. Traditional techniques such as PET/CT pose radiation hazards, whereas optical imaging poses challenges in maintaining both depth penetration and high resolution. Moreover, very few longitudinal studies have been performed for drug candidates for different symptoms. Leveraging non-ionizing photoacoustic tomography for deep tissue imaging, we developed a spatiotemporally resolved clearance pathway tracking (SRCPT) method, providing unprecedented insights into drug clearance dynamics within vital organs. SRCPT addresses challenges like laser fluence attenuation, enabling dynamic visualization of drug clearance pathways and essential parameter extraction. We employed a novel frequency component selection based synthetic aperture focusing technique (FCS-SAFT) with respiratory-artifacts-free weighting factors to enhance three-dimensional imaging resolutions. Inspired by this, we investigated the clearance pathway of a clinical drug, mitoxantrone, revealing reduced liver clearance when hepatic function is impaired. Furthermore, immunoglobulin G clearance analysis revealed significant differences among mice with varying renal injury degrees. The accuracy of our method was validated using a double-labeled probe [68Ga]DFO-IRDye800CW, showing a strong positive correlation between SRCPT and PET. We believe that this powerful SRCPT promises precise mapping of drug clearance pathways and enhances diagnosis and treatment of liver and kidney-related diseases.

Collision mapping enables accurate localization of the slow pathway in atrioventricular nodal reentrant tachycardia

Abstract Introduction Atrioventricular Node Reentry Tachycardia (AVNRT) is commonly treated with catheter ablation, with high success and low complication rates. However a small risk of complete AV block still remains. Recent studies could show that using collision map is a feasible approach in mapping the slow pathway. The aim of the current study is to show a reduction of the total ablation points needed to modify/ablate the slow pathway using the collision mapping compared to conventional 3D mapping guided slow pathway modulation/ablation and therefore minimize the risk of a complete heart block. Methods Patients diagnosed with AVNRT (n = 24) underwent collision mapping for slow pathway identification, compared to a historical cohort (n=76) using conventional mapping. For collision mapping a dynamic visualization of atrial propagation during sinus rhythm was condcted. Two propagation fronts were observed to collide at the triangle of Koch, one propagation through the slow and one through the fast pathway. The designated collision zone was considered ideal for catheter ablation (Figure 1). The endpoint after ablation in both groups was non-inducibility of AVNRTs. Results After propensity score matching the collision mapping group needed significantly fewer ablation impulses (2 (IQR 1 - 5) vs. 6 (IQR 2,5 - 11,5) p = 0.002) and shorter ablation time (p < 0.001) compared to the conventional group (Table 1). No significant differences were observed in procedure time (p = 0.209), fluoroscopy time (p = 0.296), or dose area product (p = 0.391). Conclusion Collision Mapping in AVNRT ablation showed a notable reduction in ablation points, suggesting enhanced procedural efficiency and safety.Figure 1Table1

VisConductor: Affect-Varying Widgets for Animated Data Storytelling in Gesture-Aware Augmented Video Presentation

Augmented video presentation tools provide a natural way for presenters to interact with their content, resulting in engaging experiences for remote audiences, such as when a presenter uses hand gestures to manipulate and direct attention to visual aids overlaid on their webcam feed. However, authoring and customizing these presentations can be challenging, particularly when presenting dynamic data visualization (i.e., animated charts). To this end, we introduce VisConductor, an authoring and presentation tool that equips presenters with the ability to configure gestures that control affect-varying visualization animation, foreshadow visualization transitions, direct attention to notable data points, and animate the disclosure of annotations. These gestures are integrated into configurable widgets, allowing presenters to trigger content transformations by executing gestures within widget boundaries, with feedback visible only to them. Altogether, our palette of widgets provides a level of flexibility appropriate for improvisational presentations and ad-hoc content transformations, such as when responding to audience engagement. To evaluate VisConductor, we conducted two studies focusing on presenters (𝑁=11) and audience members (𝑁=11). Our findings indicate that our approach taken with VisConductor can facilitate interactive and engaging remote presentations with dynamic visual aids. Reflecting on our findings, we also offer insights to inform the future of augmented video presentation tools.

Enhanced visualization of quench dynamics of HTS coils through embedded and distributed fiber optical sensing

The development of high-temperature superconducting (HTS) coils and magnets marks a significant advancement across various fields. To ensure their safety and reliability, thorough performance testing under extreme multi-field conditions is essential. This study introduces an innovative method that utilizes embedded and distributed fiber optical sensing (ED-FOS) in HTS pancake coils for precise real-time thermal monitoring and quench visualization. Incorporated during the manufacturing process, the ED-FOS sensors offer comprehensive and distributed measurements of thermal behavior during quenches. Our experiments demonstrate the ED-FOS effectively detects quenches caused by overcurrent and thermal issues. Unlike conventional techniques that rely on global voltage or magnetic field assessments, ED-FOS provides high spatiotemporal resolution and enhances temperature sensitivity. This capability allows for accurate, real-time identification and tracking of quench locations, which is essential for evaluating reliability and safety, as well as for improving performance. This advancement in HTS coil technology paves the way for wider application in critical fields and significantly enhances the design and management of superconducting coils.

Multicore memristor from electrically readable nanoscopic racetracks.

The manipulation and detection of mobile domain walls in nanoscopic magnetic wires underlies the development of multibit memories. The studies of such domain walls have focused on macroscopic wires that allow for optical detection by using magneto-optic effects. In this study, we demonstrated the electrical tracking with a spatial resolution of better than 40 nm of multiple mobile domain walls in nanoscopic racetracks, using a set of anomalous Hall detectors integrated into the racetracks. Electrical time-series signals from the Hall detectors allow for the static and dynamic phase space visualization of the dynamics of a domain wall or multiple domain walls that can be described by a multicore memristor model. The domain wall dynamics and stochasticity can be controlled in racetracks even to deep submicron dimensions.

Brachial and Lumbosacral Plexopathies.

The brachial and lumbosacral plexuses are complex neural structures that transmit sensory, motor, and autonomic information between the spinal cord and the extremities. Plexus disorders can be particularly disabling because lesions in the plexus usually affect large groups of nerve fibers originating from several spinal levels. Electrodiagnostic studies are often required to confirm a plexus lesion and determine the extent of injury and prognosis. Magnetic resonance is the imaging modality of choice for detecting intrinsic nerve abnormalities; recently, high-resolution ultrasound has emerged as an alternative method for dynamic evaluation and visualization of internal nerve architecture. Once a plexopathy is confirmed, the list of possible etiologies is relatively limited and includes traumatic and nontraumatic causes. Treatment relies on symptom management and physical rehabilitation unless a treatable underlying condition is found. Surgical approaches, including nerve grafts or tendon transfers, may improve limb function when spontaneous recovery is suboptimal.

Experimental Optimization of the SG6043 Airfoil for Horizontal Axis Wind Turbines Using Schmitz Equations

This study presents the experimental optimization of the SG6043 airfoil for horizontal axis wind turbines (HAWTs) using the Schmitz equation, focusing on enhancing power output and elucidating the surface flow structure. Two blade models, M1 (conventional) and M2 (optimized), were designed and tested at rotational speeds of 400 rpm and 600 rpm across a range of tip speed ratios (TSR). The M2 model, optimized using Schmitz equations, demonstrated significantly improved performance compared to the M1 model at both rotational speeds. At 400 rpm, the maximum power coefficient (CP) for M1 was 0.274, while M2 reached 0.419, indicating a 52.91% improvement. At 600 rpm, M1 achieved a maximum CP of 0.293, whereas M2 attained 0.458, representing a 56.31% enhancement. The M2 model also showed superior performance at higher TSRs, with the highest percentage increase in CP recorded at 4.9 TSR, reaching 574.54%. Additionally, dynamic surface oil-flow visualization experiments were conducted to examine flow behavior on the blade surfaces. Results indicated better flow attachment in the M2 blade due to its optimized twist angle and chord length, particularly in the mid-section, leading to delayed flow separation. The reattachment observed on the suction side of the M2 model, following the laminar separation bubble (LSB), which was absent in the M1, contributed to its higher aerodynamic efficiency and overall power performance. These findings confirm that the optimized SG6043 airfoil design, guided by Schmitz equations, offers significant improvements in HAWT performance, particularly under varying operational conditions.

Real-Time Imaging Assessment of Stress-Induced Vascular Inflammation Using Heartbeat-Synchronized Motion Compensation.

Chronic mental stress accelerates atherosclerosis through complicated neuroimmune pathways, needing for advanced imaging techniques to delineate underlying cellular mechanisms. While histopathology, ex vivo imaging, and snapshots of in vivo images offer promising evidence, they lack the ability to capture real-time visualization of blood cell dynamics within pulsatile arteries in longitudinal studies. An electrically tunable lens was implemented in intravital optical microscopy, synchronizing the focal plane with heartbeats to follow artery movements. ApoE-/- mice underwent 2 weeks of restraint stress before baseline imaging followed by 2 weeks of stress exposure in the longitudinal imaging, while nonstressed mice remained undisturbed. The progression of vascular inflammation was assessed in the carotid arteries through intravital imaging and histological analyses. A 4-fold reduction of motion artifact, assessed by interframe SD, and an effective temporal resolution of 25.2 Hz were achieved in beating murine carotid arteries. Longitudinal intravital imaging showed chronic stress led to a 6.09-fold (P=0.017) increase in myeloid cell infiltration compared with nonstressed mice. After 3 weeks, we observed that chronic stress intensified vascular inflammation, increasing adhered myeloid cells by 2.45-fold (P=0.031), while no significant changes were noted in nonstressed mice. Microcirculation imaging revealed increased circulating, rolling, and adhered cells in stressed mice's venules. Histological analysis of the carotid arteries confirmed the in vivo findings that stress augmented plaque area, myeloid cell and macrophage accumulation, and necrotic core volume while reducing fibrous cap thickness indicating accelerated plaque formation. We visualized the 3-dimensional structure of the carotid artery and 4-dimensional dynamics of the venules in the cremaster muscle. Dynamic focusing motion compensation intravital microscopy enabled subcellular resolution in vivo imaging of blood cell dynamics in beating arteries under chronic restraint stress in real time. This novel technique emphasizes the importance of advanced in vivo imaging for understanding cardiovascular disease.