Versatile Imaging Research Articles

Radiological features of the carpometacarpal joint in hamate body fracture on anteroposterior hand X-ray image

BackgroundHamate body fractures are rare and can be overlooked due to the diagnostic difficulty on X-ray. We focused on the carpometacarpal joint space width (CMC-JSW) and examined the features of this fracture on versatile anteroposterior hand X-ray images, initially taken for hand trauma cases. MethodsTen patients (nine males and one female; mean age, 34 years) diagnosed with a hamate body fracture on CT in our facility between January 2013 and October 2023 were included in this study. The fracture types were dorsal type in six cases and shear type in four cases. The 4th and 5th CMC-JSWs were measured on anteroposterior hand radiography and CT sagittal sections. If the metacarpal and hamate bones appeared to overlap due to depression, the width was expressed as a negative value (overlap sign). CMC-JSWs were compared between the types or unaffected sides using the Mann–Whitney U test. The correlation with X-ray and CT measurements was analyzed using the Spearman test. ResultsThe average CMC-JSWs (IV: −0.52 mm, V: −1.33 mm) in hamate body fractures on X-ray were minus and these values were significantly lower than unaffected side (IV: 1.51 mm, V: 1.48 mm) (p < 0.05, p < 0.001). CMC-JSW (IV: −1.64 mm) and CMC-JSW (V: −2.70 mm) of dorsal type were significantly lower than shear type (IV: 1.17 mm, V: 0.72 mm) (p < 0.05). CMC-JSW values on X-ray were positively correlated with those on CT (IV: ρ = 0.796, V: ρ = 0927). ConclusionsDecreased CMC-JSW, correlated with CT sagittal sections, and “overlap sign” on anteroposterior hand X-ray were possible radiological characteristics of hamate body fracture. Care should be taken to avoid overlooking shear-type fractures because of subtle CMC-JSW decreases.

A strategy for achieving optimisation of radiological protection in digital radiology proposed by ICRP

Radiology is now predominantly a digital medium and this has extended the flexibility, efficiency and application of medical imaging. Achieving the full benefit of digital radiology requires images to be of sufficient quality to make a reliable diagnosis for each patient, while minimising risks from radiation exposure, and so involves a careful balance between competing objectives. When an optimisation programme is undertaken, a knowledge of patient doses from surveys can be valuable in identifying areas needing attention. However, any dose reduction measures must not degrade image quality to the extent that it is inadequate for the clinical purpose. The move to digital imaging has enabled versatile image acquisition and presentation, including multi-modality display and quantitative assessment, with post-processing options that adjust for optimal viewing. This means that the appearance of an image is unlikely to give any indication when the dose is higher than necessary. Moreover, options to improve performance of imaging equipment add to its complexity, so operators require extensive training to be able to achieve this. Optimisation is a continuous rather than single stage process that requires regular monitoring, review, and analysis of performance feeding into improvement and development of imaging protocols. The ICRP is in the process of publishing two reports about optimisation in digital radiology. The first report sets out components needed to ensure that a radiology service can carry optimisation through. It describes how imaging professionals should work together as a team and explains the benefits of having appropriate methodologies to monitor performance, together with the knowledge and expertise required to use them effectively. It emphasises the need for development of organisational processes that ensure tasks are carried out. The second ICRP report deals with practical requirements for optimisation of different digital radiology modalities, and builds on information provided in earlier modality specific ICRP publications.

Intraprocedural Three-Dimensional Imaging Registration Optimizes Magnetic Resonance Imaging–Guided Focused Ultrasound and Facilitates Novel Applications

BACKGROUND AND OBJECTIVE: Transcranial magnetic resonance–guided focused ultrasound (MRgFUS) has revolutionized ablative treatment of essential tremor in recent years. However, limitations in precision targeting may account for suboptimal efficacy and significant side effects. We describe a simple intraprocedural three-dimensional image-guided lesion shaping technique that can improve overall outcomes of MRgFUS for essential tremor and facilitate expansion to novel indications. METHODS: A retrospective review of 84 consecutive MRgFUS procedures performed at Pennsylvania Hospital was performed. Comparison of patient demographics, treatment parameters, and clinical outcomes before and after implementation of this protocol was conducted. Further application of this technique in pallidotomy treatments and ablative disconnection of hypothalamic hamartoma are described. RESULTS: After implementation, the median of total number of sonications (7 vs 9, P = .001), number of therapeutic sonications (3 vs 4, P &lt; .0001), and interval time between the first and last sonication (46:10 vs 68:53 minutes, P = .0004) were significantly reduced. Patients expressed greater satisfaction of treatment (94.1% vs 82.4%, P = .018), greater global impression of change (CGI) (7 vs 6, P = .033), and reduced median number of side effects at 6 months (0 vs 1, P = .026). We also successfully implemented this protocol for novel indications. CONCLUSION: Intraprocedural lesion shaping for MRgFUS is a simple and versatile imaging protocol augmentation that improves ablation precision and can improve treatment efficacy and broader neurological application.

Going with the flow: Implementing a 4D flow MRI program at a children's hospital.

Four-dimensional phase contrast MRI (4D flow) has emerged as a versatile imaging technique for comprehensive visualization and both qualitative and quantitative assessment of cardiovascular blood flow. 4D flow is a three-dimensional, time-resolved acquisition that is gated to the cardiac cycle. 4D flow provides cardiovascular velocity and flow assessment across the volume of acquisition and yields a multitude of advanced hemodynamic parameters that help to assess the impact of cardiovascular disease on flow and vice versa, guiding the clinical and surgical management of patients with congenital and acquired heart disease. In the past, lengthy scan acquisition and complex post-processing workflows hindered 4D flow adoption into routine clinical practice. Decreasing image acquisition times and improvements in post-processing techniques have made 4D flow a clinically useful tool. The purpose of this communication is to facilitate more widespread adoption of 4D flow by describing its clinical utility, technical acquisition, optimization, and post-processing in pediatric cardiovascular imaging at our center.

Smartphone microscopy: Design and implementation of a dual magnification system

This paper presents a dual magnification smartphone microscope designed for versatile imaging, enabling low magnification for wide-field viewing and high magnification for detailed biological sample analysis and automatic control of the working distance. It offers a cost-effective alternative to traditional lab microscopes, particularly in low-resource settings. Simulated, optimized, and designed using Ansys Zemax OpticStudio, the microscope was fabricated with 3D printing technology. Dual magnification (150x and 1000x) was achieved with two optical paths, improving resolution to 1 µm at higher and 1.5 µm at lower magnifications. The design provides clear and accurate images. The smartphone integration enhances usability, allowing easy magnification switching, image capture, and potential use of diagnostic applications.

Liquid nitrogen temperature to 700 K Bi3+ thermoluminescence: Toward wide-temperature-range light dosimeters for versatile anti-counterfeiting, information storage, and x-ray imaging

Discovering light dosimeters that can function effectively from liquid nitrogen temperature to 700 K presents significant challenges. Such dosimeters facilitate a range of cutting-edge applications, including anti-counterfeiting measures at low temperature for cryo-preservation. To facilitate such discovery, stacked vacuum referred binding energy diagrams for the LiYGeO4 cluster of crystals have been first constructed. They offer a robust method for controlling both electron and hole trapping depth in the LiYGeO4 cluster of crystals. Wide temperature shifting of Bi2+ and Eu2+ thermoluminescence (TL) glow bands emerges from 200 to 500 K for LiYxLu1-xGeO4:0.01Bi3+ and LiYxLu1-xGeO4:0.01Bi3+, 0.001Eu3+, by changing x, facilitating conduction band tailoring. Wide temperature shifting of Bi4+ TL glow bands emerges from 300 to 700 K for LiYGezSi1-zO4:0.01Bi3+, by tuning z, facilitating valence band tailoring. TL glow band peaks near 135, 185, 232, and 311 K emerge in LiyNa1-yYGeO4: 0.001Bi3+. Particularly, the discovered Bi3+ or/and lanthanide modified LiYGeO4 cluster of crystals exhibit superior charge carrier storage capacity and minimal TL fading properties. For instance, the ratio of TL intensity of the optimized LiYGe0.75Si0.25O4:0.001Bi3+ to that of industrial BaFBr(I):Eu2+ is as high as ∼4. Interestingly, imaging of intense optically driven Bi3+ ultraviolet-A (UVA) luminescence has been validated in 254 nm energized LiY0.25Lu0.75GeO4:0.01Bi3+ with a 100 lux white LED illumination. Together with ZnS:Mn2+, LiTaO3:Bi3+, Sm3+, and Cs2ZrCl6:Sb3+ perovskites, the realization of wide range liquid nitrogen temperature to 700 K Bi3+ thermoluminescence in Bi3+ or/and lanthanide modified LiYGeO4 cluster of crystals with superior charge carrier storage capacity offers promising use for versatile anti-counterfeiting, information storage, and delayed x-ray imaging purposes.

Use of dual-energy x-ray absorptiometry for body composition in chronic disease management

As individuals age or contend with chronic diseases, shifts in body composition often emerge, characterized by a loss of muscle mass and an increase in fat mass, even among those with stable body weight. Both obesity and sarcopenia are key drivers of frailty, disability, and heightened morbidity and mortality. The simultaneous decline in skeletal muscle and accumulation of visceral fat can work synergistically, magnifying their detrimental effects on physical function and metabolic health. Today, dual-energy x-ray absorptiometry (DEXA) is widely recognized as one of the most versatile imaging techniques for assessing not only osteoporosis but also sarcopenia and obesity. Whole-body DEXA facilitates comprehensive analysis, offering detailed insights into fat mass, non-bone lean mass, and bone mineral content at both total and regional levels. DEXA is highly valued for its accuracy, reproducibility, speed, affordability, and low radiation exposure. Furthermore, advancements in DEXA technology and software now allow for precise estimation of visceral adipose tissue. This review underscores the clinical applications of whole-body DEXA, focusing on the use of muscle and fat mass indices in diagnosing low muscle mass, sarcopenia, and sarcopenic obesity, aligned with the latest research and guidelines.

A Versatile Image Encryption Scheme Based on Optical Hologram Technology and Chess Rules

A novel versatile image encryption scheme about hologram technology and chess rules is proposed in this paper. This scheme can encrypt images by transferring them from the spatial domain to the frequency domain. First, the theory of computer-generated holograms is applied to record phase and amplitude features by imitating the optical hologram. The original image is transformed to a hologram and the phase values of the points in the image are recorded. Second, the parameters associated with the plaintext image are obtained by a Hash function and used as the initial values of the chaotic system, and then the pawns on the board are regarded as pixel points of the image, where the movement direction of the pawns is the movement direction of the pixel points of the image. The hologram is confused by combining the pawns’ movement rules with the chaotic sequences. Finally, the encrypted image is gained through a combination of the chaotic sequence and the selective diffusion method. The test results show that the scheme is able to resist various attacks and has good security performance in encryption.

Activatable red/near-infrared aqueous organic phosphorescence probes for improved time-resolved bioimaging

Abstract Organic red/near-infrared (NIR) room-temperature phosphorescence (RTP) holds significant potential for autofluorescence-free bioimaging and biosensing due to its prolonged persistent luminescence and exceptional penetrability. However, achieving activatable red/NIR organic RTP probes with tunable emission in aqueous solution remains a formidable challenge. Here we report on aqueous organic RTP probes with red/NIR phosphorescence intensity and lifetime amplification. These probes consist of supramolecular assemblies comprising macrocyclic cucurbit[8]uril and amine-containing alkyl-bridged pyridiniums, exhibiting viscosity-activatable phosphorescence with enhanced quantum yield (up to 20%) and lifetime. Notably, by utilizing this activatable organic RTP probe, we successfully achieve two-photon imaging of lysosomal viscosity and millisecond scale time-resolved cell imaging. Moreover, intravital phosphorescence imaging using RTP probe enable the monitoring of viscosity variations in inflammatory mice, demonstrating a significantly improved signal-to-background ratio compared to fluorescence imaging. This activatable red/NIR supramolecular platform facilitates versatile high-resolution phosphorescence imaging for in vivo tracking of specific biomarkers and physiological events.

Wide-band X-ray and gamma-ray imaging for clinical application; visualization of pharmacokinetics in targeted alpha therapy

Targeted alpha therapy (TAT) is a nuclear medicine therapy which administers alpha-emitting drugs and causes localized and effective damage to tumors. To investigate whether the drugs accumulate in the tumor correctly, in-situ visualization of the distribution of the drugs is essential. Visualization of alpha-emitting drugs using X-rays and gamma rays emitted simultaneously with the alpha-rays is suggested in theranostics, a combination of therapy and diagnostics. Since various X-rays and gamma rays ranging from a few tens of keV to several MeV are emitted from the alpha-emitting radionuclides, wide-band X-ray and gamma-ray imagers are required for a more versatile visualization of alpha-emitting drugs. Therefore, we developed a hybrid Compton camera (HCC) to visualize broadband X-rays and gamma rays. In this study, we visualized representative alpha-emitting drugs, Ra-223 dichloride and At-211 NaAt, using HCC. Ra-223 dichloride in the human body was imaged using gamma rays around 300 keV, and in-vivo imaging of At-211 NaAt in a mouse was performed using 79-keV X-rays. This study demonstrates the potential of HCC as a versatile imaging device for alpha-emitting drugs.

Unsupervised Denoising in Spectral CT: Multi-Dimensional U-Net for Energy Channel Regularisation

Spectral Computed Tomography (CT) is a versatile imaging technique widely utilized in industry, medicine, and scientific research. This technique allows us to observe the energy-dependent X-ray attenuation throughout an object by using Photon Counting Detector (PCD) technology. However, a major drawback of spectral CT is the increase in noise due to a lower achievable photon count when using more energy channels. This challenge often complicates quantitative material identification, which is a major application of the technology. In this study, we investigate the Noise2Inverse image denoising approach for noise removal in spectral computed tomography. Our unsupervised deep learning-based model uses a multi-dimensional U-Net paired with a block-based training approach modified for additional energy-channel regularization. We conducted experiments using two simulated spectral CT phantoms, each with a unique shape and material composition, and a real scan of a biological sample containing a characteristic K-edge. orangeMeasuring the peak signal-to-noise ratio (PSNR) and structural similarity index (SSIM) for the simulated data and the contrast-to-noise ratio (CNR) for the real-world data, our approach not only outperforms previously used methods—namely the unsupervised Low2High method and the total variation-constrained iterative reconstruction method—but also does not require complex parameter tuning.

Synchrotron X-ray imaging of soft biological tissues - principles, applications and future prospects.

Synchrotron-based tomographic phase-contrast X-ray imaging (SRµCT or SRnCT) is a versatile isotropic three-dimensional imaging technique that can be used to study biological samples spanning from single cells to human-sized specimens. SRµCT and SRnCT take advantage of the highly brilliant and coherent X-rays produced by a synchrotron light source. This enables fast data acquisition and enhanced image contrast for soft biological samples owing to the exploitation of phase contrast. In this Review, we provide an overview of the basics behind the technique, discuss its applications for biologists and provide an outlook on the future of this emerging technique for biology. We introduce the latest advances in the field, such as whole human organs imaged with micron resolution, using X-rays as a tool for virtual histology and resolving neuronal connections in the brain.

Kilohertz volumetric imaging of in-vivo dynamics using squeezed light field microscopy.

Volumetric functional imaging of transient cellular signaling and motion dynamics poses a significant challenge to current microscopy techniques, primarily due to limitations in hardware bandwidth and the restricted photon budget within short exposure times. In response to this challenge, we present squeezed light field microscopy (SLIM), a computational imaging method that enables rapid detection of high-resolution three-dimensional (3D) light signals using only a single, low-format camera sensor area. SLIM pushes the boundaries of 3D optical microscopy, achieving over one thousand volumes per second across a large field of view of 550 μm in diameter and 300 μm in depth with a spatial resolution of 3.6 μm laterally and 6 μm axially. Using SLIM, we demonstrated blood cell velocimetry across the embryonic zebrafish brain and in a free-moving tail exhibiting high-frequency swinging motion. The millisecond temporal resolution also enables accurate voltage imaging of neural membrane potentials in the leech ganglion, and in the hippocampus of behaving mice. These results collectively establish SLIM as a versatile and robust imaging tool for high-speed microscopy applications.

MRI-Seed-Wizard: Combining Deep Learning Algorithms with Magnetic Resonance Imaging Enables Advanced Seed Phenotyping.

Evaluation of relevant seed traits is an essential part of most plant breeding and biotechnology programs. There is need for non-destructive, three-dimensional assessment of the morphometry, composition, and internal features of seeds. Here, we introduced a novel tool, MRI-Seed-Wizard, which integrates deep learning algorithms with non-invasive magnetic resonance imaging (MRI) for its use in the new domain - plant MRI. The tool enabled in vivo quantification of 23 grain traits, including volumetric parameters of inner seed structure. Several of these features cannot be assessed using conventional techniques, including X-ray computed tomography. MRI-Seed-Wizard was designed to automate the manual processes of identifying, labeling, and analyzing digital MRI data. We further provide advanced MRI protocols that allow the evaluation of multiple seeds simultaneously to increase throughput. The versatility of MRI-Seed-Wizard in seed phenotyping was demonstrated for wheat (Triticum aestivum) and barley (Hordeum vulgare) grains, and is applicable to a wide range of crop seeds. Thus, artificial intelligence, combined with the most versatile imaging modality - MRI, opens up new perspectives in seed phenotyping and crop improvement.

Diagnostic Value of Cardiac Magnetic Resonance Imaging (CMRI) in the Evaluation of Arrhythmia and Cardiomyopathy.

Background Cardiac magnetic resonance imaging (CMRI) is now the gold standard noninvasive method for evaluating ventricular volume, mass, and ejection fraction - crucial indicators for diagnosing and prognosticating heart failure. Renowned for its accuracy and reliability, CMRI utilizes advanced techniques, such as late gadolinium enhancement and T1 and T2 mapping, to produce three-dimensional images with high spatial and temporal resolution, all without ionizing radiation. One of CMRI's key strengths lies in its capacity to characterize myocardial tissue, providing valuable diagnostic and prognostic insights into underlying cardiac conditions. Recent studies underscore CMRI's feasibility, safety, and emerging cost-effectiveness in selected clinical settings. However, the complex etiologies of arrhythmias and cardiomyopathies, marked by varied structural and functional myocardial changes, continue to present diagnostic challenges. While CMRI's role is expanding, comprehensive observational studies are essential to fully elucidate its diagnostic utility in these cases. Aim This study aims to assess the utility of CMRI in the etiological diagnosis of heart failure and arrhythmias, addressing current knowledge gaps and contributing to clinical practice insights. Overall, CMRI is highlighted as a versatile and effective imaging modality for evaluating diverse aspects of cardiac health. Methods This single-center, hospital-based, non-randomized, prospective observational study was conducted in the cardiology department of Pushpawati Singhania Research Institute (PSRI) Hospital in New Delhi, India, from December 2019 to December 2021. It included patients presenting with arrhythmias, cardiomyopathies, unexplained left ventricular (LV) dysfunction, suspected arrhythmogenic right ventricular dysplasia, myocarditis, hypertrophic cardiomyopathy, as well as infiltrative and granulomatous diseases. Only patients who provided consent for cardiac MRI were included. Cardiac MRI tests were performed and analyzed for all participants to evaluate their diagnostic utility across these conditions. Results The study involved 100 participants, with 75% diagnosed with cardiomyopathy and 25% with arrhythmia. Among the cardiomyopathy cases, 24 patients (32%) had ischemic cardiomyopathy, while 51 patients (68%) had nonischemic cardiomyopathy. LV dysfunction was noted in 65 patients (86.6%), with a viable scar present in three patients (4%) and myocarditis diagnosed in four patients (5.3%). Chamber dilatation was observed in 13 patients (52%). In the arrhythmia group, six patients (24%) exhibited prolonged native T1, while eight patients (32%) had normal cardiac MRI results. These findings highlight the significant utility of cardiac MRI in diagnosing and characterizing various cardiac conditions within the study population. Conclusions The study found that cardiac MRI exhibits a strong positive predictive value in diagnosing various types of cardiomyopathies. This suggests that when CMRI identifies cardiomyopathy, it is likely to provide an accurate diagnosis, thereby enhancing its utility in clinical practice for effective patient management and treatment planning.

Single-molecule detection and super-resolution imaging with a portable and adaptable 3D-printed microscopy platform (Brick-MIC).

Over the past decades, single-molecule and super-resolution microscopy have advanced and represent essential tools for life science research. There is, however, a growing gap between the state of the art and what is accessible to biologists, biochemists, medical researchers, or labs with financial constraints. To bridge this gap, we introduce Brick-MIC, a versatile and affordable open-source 3D-printed microspectroscopy and imaging platform. Brick-MIC enables the integration of various fluorescence imaging techniques with single-molecule resolution within a single platform and exchange between different modalities within minutes. We here present variants of Brick-MIC that facilitate single-molecule fluorescence detection, fluorescence correlation spectroscopy, time-correlated single-photon counting and super-resolution imaging (STORM and PAINT). Detailed descriptions of the hardware and software components, as well as data analysis routines, are provided, to allow non-optics specialists to operate their own Brick-MIC with minimal effort and investments. We foresee that our affordable, flexible, and open-source Brick-MIC platform will be a valuable tool for many laboratories worldwide.

A Comprehensive Review: Versatile Imaging Probe Based on Chemical Materials for Biomedical Applications.

Imaging probe and contrast agents play significant role in combating cancer. Based on special chemical materials, imaging probe can convert cancer symptoms into information-rich images with high sensitivity and signal amplification, accompanying with detection, diagnosis, drug delivery and treatment. In the paper, some inorganic and organic chemical materials as imaging probe, including Ultrasound imaging (US), Optical imaging (OP), Photoacoustic imaging (PA), X-ray Computed Tomography (CT), Magnetic Resonance imaging (MRI), Radionuclide imaging (RNI) probe, as well as multi-modality imaging probe for diagnosis and therapy of tumour were introduced. The sophisticated and comprehensive chemical materials as imaging probe were highlighted in detail. Meanwhile, the advantages and disadvantages of the imaging probe were compared. In order to provide some reference and help researchers for construction imaging probe for tumour diagnosis and treatment, it attempts to exhaustively cover the whole field. Finally, the prospect and challenge for imaging probe were discussed.

Prominence of curcumin in anticancer strategies and innovation in bioimaging

Curcumin (CUR), a natural polyphenol derived from the rhizomes of Curcuma longa (turmeric), has garnered significant attention in recent years for its multifaceted potential in anticancer therapy and bio-imaging applications. This review article comprehensively explores the diverse anticancer properties of CUR, encompassing its potent anti-inflammatory, antioxidant, and anti-proliferative effects, as well as its ability to induce apoptosis and inhibit angiogenesis, making it a promising candidate in the fight against cancer. In addition to its therapeutic potential, CUR's unique physicochemical properties have enabled its utility as a versatile imaging agent for various bio-imaging modalities, including fluorescence imaging, magnetic resonance imaging (MRI), and positron emission tomography (PET). We investigate the molecular mechanisms underlying CUR's bio-imaging capabilities and discuss its various applications in cancer diagnosis, monitoring treatment responses, and elucidating biological processes. This comprehensive review provides valuable insights into the dual role of CUR as an anticancer agent and a bio-imaging tool, elucidate its potential in the development of novel cancer therapies and diagnostic approaches. The amalgamation of CUR's bio-imaging and therapeutic properties suggests its future as a pivotal player in personalized medicine and precision oncology.

Ultraplex microscopy: versatile highly-multiplexed molecular labeling and imaging across scale and resolution.

The molecular organization of cells and tissue is challenging to study due to the inefficiency of multiplexed molecular labeling methods and the limited options for combining microscopy modalities in a single specimen, especially when high spatial resolution is needed. Here we describe ultraplex microscopy, which combines serial multiplexing, ultrathin sectioning, and reversible embedding to circumvent incompatibilities between labeling and imaging techniques, enhance resolution, and expand multiplexing capacity within and across modalities. Samples can be labeled with antibodies, RNA probes, and tissue stains for imaging by brightfield, epifluorescence, super-resolution, and electron microscopy without specialized reagents or materials. We demonstrate applications in brain tissue including molecular profiling of single cells and axonal boutons, high-resolution molecular colocalization, and correlative imaging of fluorescent proteins with confocal and ultraplex microscopy. The power and versatility of ultraplex microscopy will be valuable in addressing currently intractable experimental questions in many systems and contexts.

Fluorescence Lifetime Multiplexing (FLEX) for simultaneous high dimensional spatial biology in 3D

Immunohistochemistry is a crucial method for detecting specific proteins within tissue samples, yet constrained to one biomarker per tissue section. Multiplexed immunofluorescence, while allowing simultaneous visualization of multiple proteins, faces limitations in the number of simultaneous fluorescent labels due to spectral overlap. Although cyclic immunofluorescence techniques have successfully broadened antibody staining capacities in a single tissue sample, they are plagued by time-consuming and labor-intensive procedures, sample degradation risks, and inability to scale beyond thin sections. In this study, we introduce the use of 3D confocal Fluorescence Lifetime Imaging Microscopy as a high-throughput, multiplexed immunofluorescence platform that can differentiate 11 or more biomarkers in 3D tissue volumes. Leveraging both spectral and lifetime information, this approach allows for practical spatial biology in thin sections that can readily scale to larger volumes of tissue. We believe that this highly multiplexed and versatile biomarker imaging platform will significantly expedite cancer research and enable new translational approaches in the future.