No Time to Lose: Enabling Real-Time Fluorescence Lifetime Imaging on Resource-constrained FPGAs Through Efficient Scheduling

Опубликована: Фев. 26, 2025

Fluorescence lifetime imaging (FLI) is a widely used technique in the biomedical field for measuring decay times of fluorescent molecules, providing insights into metabolic states, protein interactions, and ligand-receptor bindings [1, 5]. However, its broader application fast biological processes, such as dynamic activity monitoring, clinical use, guided surgery, limited by long data acquisition computationally demanding processing. While deep learning has reduced post-processing times, time-resolved remains bottleneck real-time applications. To address this,we propose method to achieve near realtime FLI using an FPGA-based hardware accelerator. Specifically,we implemented GRU-based sequence-to-sequence (Seq2Seq) model [3] on FPGA board compatible with cameras. The GRU balances accurate processing resource constraints FPGAs, which have DSP units BRAM. memory computational resources require efficient scheduling operations allocation deploy models low-latency applications.We these challenges STOMP, queue-based discrete-event simulator that automates optimizes task management [4]. By integrating Seq2Seq compressed version, called Seq2SeqLite [2], generated through knowledge distillation (KD), we were able process multiple pixels parallel, reducing latency compared sequential We explore various levels parallelism optimal balance between performance utilization. Our results indicate proposed techniques achieved 17.7x 52.0x speedup over manual model, respectively. Furthermore, 98% reduction parameters model.

Язык: Английский

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Persistent Luminescence Lifetime-Based Near-Infrared Nanoplatform via Deep Learning for High-Fidelity Biosensing of Hypochlorite

Analytical Chemistry, Год журнала: 2024, Номер 96(18), С. 7240 - 7247

Опубликована: Апрель 25, 2024

In light of deep tissue penetration and ultralow background, near-infrared (NIR) persistent luminescence (PersL) bioprobes have become powerful tools for bioapplications. However, the inhomogeneous signal attenuation may significantly limit its application precise biosensing owing to absorption scattering. this work, a PersL lifetime-based nanoplatform via learning was proposed high-fidelity bioimaging in vivo. The imaging network (PLI-Net), which consisted 3D-deep convolutional neural (3D-CNN) system, logically constructed accurately extract lifetime feature from profile intensity-based decay images. Significantly, NIR nanomaterials represented by Zn1+xGa2–2xSnxO4: 0.4 % Cr (ZGSO) were precisely adjusted over their lifetime, enabling with high-contrast signals. Inspired adjustable reliable ZGSO NPs, proof-of-concept further developed showed exceptional analytical performance hypochlorite detection resonance energy transfer process. Remarkably, on merits dependable anti-interference lifetimes, nanoprobe provided highly sensitive accurate both endogenous exogenous hypochlorite. This breakthrough opened up new way development complex matrix systems.

Язык: Английский

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In Vivo Lifetime Imaging of the Internal O₂ Dynamics in Corals with near-Infrared-Emitting Sensor Nanoparticles

ACS Sensors, Год журнала: 2024, Номер 9(9), С. 4671 - 4679

Опубликована: Авг. 23, 2024

Mapping of O2 with luminescent sensors within intact animals is challenging due to attenuation excitation and emission light caused by tissue absorption scattering as well interfering background fluorescence. Here we show the application sensor nanoparticles (∼50–70 nm) composed indicator platinum(II) tetra(4-fluoro)phenyltetrabenzoporphyrin (PtTPTBPF) immobilized in poly(methyl methacrylate-co-methacrylic acid) (PMMA-MA). We injected into gastrovascular system colony fractions reef-building tropical corals that harbor photosynthetic microalgae their tissues. The are excited red LED (617 emit near-infrared (780 nm), which enhances transmission through biological materials. This enabled us map internal concentration via time-domain luminescence lifetime imaging outer layers across several coral polyps flowing seawater. After injection, dispersed for hours. While intensity showed some local aggregation particles, a more homogeneous distribution larger area colony. Local stimulation symbiont photosynthesis induced oxygenation illuminated areas formation lateral gradients toward surrounding respiring tissues, were dissipated rapidly after onset darkness. Such measurements key improving our understanding how regulate chemical microenvironment metabolic activity, they affected environmental stress such ocean warming, acidification, deoxygenation. Our experimental approach can also be adapted vivo other natural systems biofilms, plant animal organoids cell constructs, where conditions relevant high optical density

Язык: Английский

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Detecting inflammation in the diabetic mice with a fluorescence lifetime-based probe

Analytica Chimica Acta, Год журнала: 2022, Номер 1221, С. 340104 - 340104

Опубликована: Июнь 20, 2022

Язык: Английский

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Oxygen Measurement in Microdevices

Annual Review of Analytical Chemistry, Год журнала: 2022, Номер 15(1), С. 221 - 246

Опубликована: Июнь 13, 2022

Oxygen plays a fundamental role in respiration and metabolism, quantifying oxygen levels is essential many environmental, industrial, research settings. Microdevices facilitate the study of dynamic, oxygen-dependent effects real time. This review organized around key needs for measurement microdevices, including integrability into microfabricated systems; sensor dynamic range sensitivity; spatially resolved measurements to map over two- or three-dimensional regions interest; compatibility with multimodal multianalyte measurements. After brief overview biological readouts oxygen, followed by types that have been implemented microscale devices sensing mechanisms, this presents select recent applications organs-on-chip vitro models new capabilities enabling microscopy, bioprocess manufacturing, pharmaceutical industries. With advancement multiplexed, interconnected sensors instruments integration industry workflows, intelligent microdevice-sensor systems will further impact environmental science, medicine.

Язык: Английский

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In vivo quantitative FRET small animal imaging: Intensity versus lifetime-based FRET

Biophysical Reports, Год журнала: 2023, Номер 3(2), С. 100110 - 100110

Опубликована: Май 9, 2023

Förster resonance energy transfer (FRET) microscopy is used in numerous biophysical and biomedical applications to monitor inter- intramolecular interactions conformational changes the 2-10 nm range. FRET currently being extended vivo optical imaging, its main application quantifying drug-target engagement or drug release animal models of cancer using organic dye nanoparticle-labeled probes. Herein, we compared quantification intensity-based (sensitized emission analysis with three-cube approach an IVIS imager) macroscopic fluorescence lifetime (MFLI) a custom system time-gated-intensified charge-coupled device, for small imaging. The analytical expressions experimental protocols required quantify product

Язык: Английский

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Fluorescence Lifetime Imaging for Quantification of Targeted Drug Delivery in Varying Tumor Microenvironments

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Янв. 15, 2024

ABSTRACT Rationale Trastuzumab (TZM) is a monoclonal antibody that targets the human epidermal growth factor receptor (HER2) and clinically used for treatment of HER2-positive breast tumors. However, tumor microenvironment can limit access TZM to HER2 across whole thereby compromise TZM’s therapeutic efficacy. An imaging methodology non-invasively quantify binding TZM-HER2, which required action, distribution within tumors with varying microenvironments much needed. Methods We performed near-infrared (NIR) fluorescence lifetime (FLI) Forster Resonance Energy Transfer (FRET) measure TZM-HER2 binding, using in vitro microscopy vivo widefield macroscopy, overexpressing ovarian cancer cells xenografts, respectively. Immunohistochemistry was validate results. Results NIR FLI FRET data show variations intracellular bound AU565 tumor-passaged XTM cell lines comparison SKOV-3 cells. Macroscopy (MFLI) display reduced compared tumors, as validated by ex immunohistochemistry. Moreover, AU565/XTM xenografts different amounts distributions TME components, such collagen vascularity. Therefore, these results suggest are refractory delivery due their disrupted vasculature increased content. Conclusion Our study demonstrates powerful analytical tool monitor drug both cultures live systems. Especially, MFLI unique modality directly target engagement potential elucidate role efficacy intact xenografts.

Язык: Английский

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NAD(P)H fluorescence lifetime imaging of live intestinal nematodes reveals metabolic crosstalk between parasite and host

Scientific Reports, Год журнала: 2022, Номер 12(1)

Опубликована: Май 4, 2022

Abstract Infections with intestinal nematodes have an equivocal impact: they represent a burden for human health and animal husbandry, but, at the same time, may ameliorate auto-immune diseases due to immunomodulatory effect of parasites. Thus, it is key understand how arrive persist in their luminal niche interact host over long periods time. One basic mechanism governing parasite cellular tissue functions, metabolism, has largely been neglected study nematode infections. Here we use NADH (nicotinamide adenine dinucleotide) NADPH dinucleotide phosphate) fluorescence lifetime imaging explanted murine duodenum infected natural Heligmosomoides polygyrus define link between general metabolic activity possible pathways tissue, during acute infection. In both healthy intestine, energy effectively produced, mainly via resembling oxidative phosphorylation/aerobic glycolysis features. contrast, shift production from balanced fast anaerobic glycolysis-like effective phosphorylation-like pathways, towards back different life cycle phases submucosa versus lumen. Additionally, found increased oxidase (NOX) enzymes-dependent burst as compared which was mirrored by similar defense reaction We expect that, here presented application NAD(P)H-FLIM live tissues constitutes unique tool shifts host-parasite crosstalk, various parasitic

Язык: Английский

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Neural Networks Push the Limits of Luminescence Lifetime Nanosensing

Advanced Materials, Год журнала: 2023, Номер 35(52)

Опубликована: Окт. 3, 2023

Abstract Luminescence lifetime‐based sensing is ideally suited to monitor biological systems due its minimal invasiveness and remote working principle. Yet, applicability limited in conditions of low signal‐to‐noise ratio (SNR) induced by, e.g., short exposure times presence opaque tissues. Herein this limitation overcome by applying a U‐shaped convolutional neural network (U‐NET) improve luminescence lifetime estimation under extremely SNR. Specifically, the prowess U‐NET showcased context thermometry, achieving more precise thermal readouts using Ag 2 S nanothermometers. Compared traditional analysis methods decay curve fitting integration, can extract average lifetimes precisely consistently regardless SNR value. The improvement achieved performance demonstrated with two experiments characterized extreme measurement conditions: monitoring free‐falling droplets, transients suspended droplets through an medium. These results broaden fields including vivo experimentation microfluidics, while, hopefully, spurring further research on implementation machine learning (ML) sensing.

Язык: Английский

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Two-Photon Fluorescence Microscopy and Applications in Angiogenesis and Related Molecular Events

Tissue Engineering Part B Reviews, Год журнала: 2021, Номер 28(4), С. 926 - 937

Опубликована: Сен. 20, 2021

The role of angiogenesis in health and disease have gained considerable momentum recent years. Visualizing angiogenic patterns associated events surrounding vascular beds response to therapeutic laboratory-grade biomolecules has become a commonplace regenerative medicine the biosciences. To achieve high-quality imaging for elucidating molecular mechanisms angiogenesis, two-photon excitation fluorescence (2PEF) microscopy, or multiphoton microscopy is increasingly utilized scientific investigations. 2PEF microscope confers several distinct advantages over other techniques—for observation in-depth, three-dimensional vascularity variety tissue formats, including fixed specimens vivo vasculature live specimens. Understanding morphological subcellular changes that occur cells tissues during will provide insights behavioral responses diseased states, advance engineering physiologically relevant models, biochemical clues design strategies. We review applicability limitations on biophysical molecular-level signatures various models. Imaging techniques strategies best practices be reviewed. Deep provides unique opportunities study real-time. In contrast cross-sectional data provided by conventional methods, enables high-resolution imaging, acquisition time, real-time visualization events, reduces number animal models used research. This different methods its application deep vitro tissues. believe current trends can transform investigation cancer research, biofabrication vascularized

Язык: Английский

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