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1.
J Biomed Opt ; 30(Suppl 1): S13703, 2025 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-39034959

RESUMO

Significance: Standardization of fluorescence molecular imaging (FMI) is critical for ensuring quality control in guiding surgical procedures. To accurately evaluate system performance, two metrics, the signal-to-noise ratio (SNR) and contrast, are widely employed. However, there is currently no consensus on how these metrics can be computed. Aim: We aim to examine the impact of SNR and contrast definitions on the performance assessment of FMI systems. Approach: We quantified the SNR and contrast of six near-infrared FMI systems by imaging a multi-parametric phantom. Based on approaches commonly used in the literature, we quantified seven SNRs and four contrast values considering different background regions and/or formulas. Then, we calculated benchmarking (BM) scores and respective rank values for each system. Results: We show that the performance assessment of an FMI system changes depending on the background locations and the applied quantification method. For a single system, the different metrics can vary up to ∼ 35 dB (SNR), ∼ 8.65 a . u . (contrast), and ∼ 0.67 a . u . (BM score). Conclusions: The definition of precise guidelines for FMI performance assessment is imperative to ensure successful clinical translation of the technology. Such guidelines can also enable quality control for the already clinically approved indocyanine green-based fluorescence image-guided surgery.


Assuntos
Benchmarking , Imagem Molecular , Imagem Óptica , Imagens de Fantasmas , Razão Sinal-Ruído , Imagem Molecular/métodos , Imagem Molecular/normas , Imagem Óptica/métodos , Imagem Óptica/normas , Processamento de Imagem Assistida por Computador/métodos
2.
Theranostics ; 14(10): 4127-4146, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38994026

RESUMO

Background: Biomarker-driven molecular imaging has emerged as an integral part of cancer precision radiotherapy. The use of molecular imaging probes, including nanoprobes, have been explored in radiotherapy imaging to precisely and noninvasively monitor spatiotemporal distribution of biomarkers, potentially revealing tumor-killing mechanisms and therapy-induced adverse effects during radiation treatment. Methods: We summarized literature reports from preclinical studies and clinical trials, which cover two main parts: 1) Clinically-investigated and emerging imaging biomarkers associated with radiotherapy, and 2) instrumental roles, functions, and activatable mechanisms of molecular imaging probes in the radiotherapy workflow. In addition, reflection and future perspectives are proposed. Results: Numerous imaging biomarkers have been continuously explored in decades, while few of them have been successfully validated for their correlation with radiotherapeutic outcomes and/or radiation-induced toxicities. Meanwhile, activatable molecular imaging probes towards the emerging biomarkers have exhibited to be promising in animal or small-scale human studies for precision radiotherapy. Conclusion: Biomarker-driven molecular imaging probes are essential for precision radiotherapy. Despite very inspiring preliminary results, validation of imaging biomarkers and rational design strategies of probes await robust and extensive investigations. Especially, the correlation between imaging biomarkers and radiotherapeutic outcomes/toxicities should be established through multi-center collaboration involving a large cohort of patients.


Assuntos
Biomarcadores Tumorais , Imagem Molecular , Neoplasias , Humanos , Neoplasias/radioterapia , Neoplasias/diagnóstico por imagem , Imagem Molecular/métodos , Animais , Biomarcadores Tumorais/metabolismo , Sondas Moleculares/química , Radioterapia/métodos , Radioterapia/efeitos adversos , Biomarcadores/metabolismo
3.
J Nanobiotechnology ; 22(1): 421, 2024 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-39014370

RESUMO

BACKGROUND: Prostate cancer (PCa) is the most prevalent cancer among males, emphasizing the critical need for precise diagnosis and treatment to enhance patient prognosis. Recent studies have extensively utilized urine exosomes from patients with cancer for targeted delivery. This study aimed to employ highly sensitive magnetic particle imaging (MPI) and fluorescence molecular imaging (FMI) to monitor the targeted delivery of an exosome-loaded platform at the tumour site, offering insights into a potential combined photothermal and magnetic thermal therapy regime for PCa. RESULTS: MPI and FMI were utilized to monitor the in vivo retention performance of exosomes in a prostate tumour mouse model. The exosome-loaded platform exhibited robust homologous targeting ability during imaging (SPIONs@EXO-Dye:66·48%±3·85%; Dye-SPIONs: 34·57%±7·55%, **P<0·01), as verified by in vitro imaging and in vitro tissue Prussian blue staining. CONCLUSIONS: The experimental data underscore the feasibility of using MPI for in vivo PCa imaging. Furthermore, the exosome-loaded platform may contribute to the precise diagnosis and treatment of PCa.


Assuntos
Exossomos , Neoplasias da Próstata , Animais , Masculino , Exossomos/metabolismo , Exossomos/química , Neoplasias da Próstata/diagnóstico por imagem , Neoplasias da Próstata/terapia , Camundongos , Humanos , Linhagem Celular Tumoral , Imagem Óptica/métodos , Modelos Animais de Doenças , Terapia Fototérmica/métodos , Imagem Molecular/métodos , Camundongos Nus
4.
Methods Mol Biol ; 2816: 129-138, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38977594

RESUMO

Phospholipase D (PLD) is an enzyme with many functions, one of which is the synthesis of phosphatidic acid (PA), a molecule with a myriad of effects on various organ systems and processes. These numerous roles make it hard to understand the true action of PA in cellular and bodily processes. Imaging PLD activity is one way to better understand the synthesis of PA and start to elucidate its function. However, many of the current imaging techniques for PLD come with limitations. This chapter presents a thorough methodology of a new imaging technique for PLD activity with clickable alcohols via transphosphatidylation (IMPACT) and Real-Time IMPACT (RT-IMPACT) that takes advantage of clickable chemistry to overcome current limitations. Using strain-promoted azide-alkyne cycloaddition (SPAAC), inverse electron-demand Diels-Alder (IEDDA), and the synthesis of various organic compounds, this chapter will explain a step-by-step procedure of how to perform the IMPACT and RT-IMPACT method(s).


Assuntos
Álcoois , Química Click , Fosfolipase D , Fosfolipase D/metabolismo , Fosfolipase D/química , Química Click/métodos , Álcoois/química , Álcoois/metabolismo , Reação de Cicloadição , Humanos , Ácidos Fosfatídicos/metabolismo , Ácidos Fosfatídicos/química , Azidas/química , Imagem Molecular/métodos , Alcinos/química
5.
J Mass Spectrom ; 59(8): e5074, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39017393

RESUMO

Mass spectrometry imaging (MSI) was developed to visualize spatial chemical information within tissues, thereby facilitating spatial multi-omic analysis. However, due to the limited spatial information provided by individual modal MSI, correlating various chemical data within tissues remains a significant challenge. In recent years, multimodal MSI has garnered considerable attention due to its ability to visualize the spatial distributions of multiple biomolecules within tissues. Among the strategies employed in this field, multimodal imaging on a single tissue section circumvents multiple issues introduced by integration of images of consecutive tissue sections. In this minireview, we provide an overview of multimodal MSI on a single tissue section, with a particular focus on the use of Matrix-Assisted Laser Desorption/Ionization-MSI for spatial multi-omic investigations that offer a comprehensive and in-depth elucidation of the biological state and activities, aiming to inspire the development of new approaches in this field.


Assuntos
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Humanos , Animais , Imagem Multimodal/métodos , Imagem Molecular/métodos
6.
Expert Rev Endocrinol Metab ; 19(4): 317-333, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38899737

RESUMO

INTRODUCTION: Molecular imaging of thyroid and parathyroid diseases has changed in recent years due to the introduction of new radiopharmaceuticals and new imaging techniques. Accordingly, we provided an clinicians-oriented overview of such techniques and their indications. AREAS COVERED: A review of the literature was performed in the PubMed, Web of Science, and Scopus without time or language restrictions through the use of one or more fitting search criteria and terms as well as through screening of references in relevant selected papers. Literature up to and including December 2023 was included. Screening of titles/abstracts and removal of duplicates was performed and the full texts of the remaining potentially relevant articles were retrieved and reviewed. EXPERT OPINION: Thyroid and parathyroid scintigraphy remains integral in patients with thyrotoxicosis, thyroid nodules, differentiated thyroid cancer and, respectively, hyperparathyroidism. In the last years positron-emission tomography with different tracers emerged as a more accurate alternative in evaluating indeterminate thyroid nodules [18F-fluorodeoxyglucose (FDG)], differentiated thyroid cancer [124I-iodide, 18F-tetrafluoroborate, 18F-FDG] and hyperparathyroidism [18F-fluorocholine]. Other PET tracers are useful in evaluating relapsing/advanced forms of medullary thyroid cancer (18F-FDOPA) and selecting patients with advanced follicular and medullary thyroid cancers for theranostic treatments (68Ga/177Ga-somatostatin analogues).


Assuntos
Imagem Molecular , Doenças das Paratireoides , Compostos Radiofarmacêuticos , Doenças da Glândula Tireoide , Humanos , Imagem Molecular/métodos , Doenças das Paratireoides/diagnóstico por imagem , Doenças da Glândula Tireoide/diagnóstico por imagem , Tomografia por Emissão de Pósitrons
8.
Biol Pharm Bull ; 47(6): 1066-1071, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38825459

RESUMO

Both nuclear and optical imaging are used for in vivo molecular imaging. Nuclear imaging displays superior quantitativity, and it permits imaging in deep tissues. Thus, this method is widely used clinically. Conversely, because of the low permeability of visible to near-IR light in living animals, it is difficult to visualize deep tissues via optical imaging. However, the light at these wavelengths has no ionizing effect, and it can be used without any restrictions in terms of location. Furthermore, optical signals can be controlled in vivo to accomplish target-specific imaging. Nuclear medicine and phototherapy have also evolved to permit targeted-specific imaging. In targeted nuclear therapy, beta emitters are conventionally used, but alpha emitters have received significant attention recently. Concerning phototherapy, photoimmunotherapy with near-IR light was approved in Japan in 2020. In this article, target-specific imaging and molecular targeted therapy utilizing nuclear medicine and optical technologies are discussed.


Assuntos
Imagem Molecular , Medicina Nuclear , Imagem Óptica , Humanos , Animais , Imagem Óptica/métodos , Imagem Molecular/métodos , Medicina Nuclear/métodos , Fototerapia/métodos , Terapia de Alvo Molecular/métodos , Neoplasias/terapia , Neoplasias/diagnóstico por imagem
9.
Radiol Imaging Cancer ; 6(4): e230186, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38847615

RESUMO

Purpose To develop a molecular breast imaging (MBI)-guided biopsy system using dual-detector MBI and to perform initial testing in participants. Materials and Methods The Stereo Navigator MBI Accessory biopsy system comprises a lower detector, upper fenestrated compression paddle, and upper detector. The upper detector retracts, allowing craniocaudal, oblique, or medial or lateral biopsy approaches. The compression paddle allows insertion of a needle guide and needle. Lesion depth is calculated by triangulation of lesion location on the upper detector at 0° and 15° and relative lesion activity on upper and lower detectors. In a prospective study (July 2022-June 2023), participants with Breast Imaging Reporting and Data System category 2, 3, 4, or 5 breast lesions underwent MBI-guided biopsy. After injection of 740 MBq technetium 99m sestamibi, craniocaudal and mediolateral oblique MBI (2-minute acquisition per view) confirmed lesion visualization. A region of interest over the lesion permitted depth calculation in the system software. Upper detector retraction allowed biopsy device placement. Specimen images were obtained on the retracted upper detector, confirming sampling of the target. Results Of 21 participants enrolled (mean age, 50.6 years ± 10.1 [SD]; 21 [100%] women), 17 underwent MBI-guided biopsy with concordant pathology. No lesion was observed at the time of biopsy in four participants. Average lesion size was 17 mm (range, 6-38 mm). Average procedure time, including preprocedure imaging, was 55 minutes ± 13 (range, 38-90 minutes). Pathology results included invasive ductal carcinoma (n = 1), fibroadenoma (n = 4), pseudoangiomatous stromal hyperplasia (n = 6), and fibrocystic changes (n = 6). Conclusion MBI-guided biopsy using a dual-head system with retractable upper detector head was feasible, well tolerated, and efficient. Keywords: Breast Biopsy, Molecular Breast Imaging, Image-guided Biopsy, Molecular Breast Imaging-guided Biopsy, Breast Cancer Clinical trial registration no. NCT06058650 © RSNA, 2024.


Assuntos
Neoplasias da Mama , Biópsia Guiada por Imagem , Imagem Molecular , Tecnécio Tc 99m Sestamibi , Humanos , Feminino , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/patologia , Pessoa de Meia-Idade , Estudos Prospectivos , Biópsia Guiada por Imagem/métodos , Biópsia Guiada por Imagem/instrumentação , Adulto , Imagem Molecular/métodos , Imagem Molecular/instrumentação , Idoso , Compostos Radiofarmacêuticos , Mama/diagnóstico por imagem
10.
J Nucl Med Technol ; 52(2): 107-114, 2024 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-38839120

RESUMO

Molecular breast imaging (MBI) is one of several options available to patients seeking supplemental screening due to mammographically dense breasts. Patient experience during MBI may influence willingness to undergo the test but has yet to be formally assessed. We aimed to assess patient comfort level during MBI, to compare MBI comfort with mammography comfort, to identify factors associated with MBI discomfort, and to evaluate patients' willingness to return for future MBI. Methods: A 10-question survey was sent by e-mail to patients undergoing MBI between August and December 2022 to obtain quantitative assessments and qualitative opinions about MBI. Results: Of 561 invited patients, 209 (37%) completed the survey and provided study consent. Their average age was 60.1 y (range, 40-81 y). Of the 209 responders, 202 (97%) were presenting for screening MBI, 195 (94%) had dense breasts, and 46 (22%) had a personal history of breast cancer. The average rating of MBI comfort was 2.9 (SD, 1.5; median, 3.0) on a 7-point scale (1 indicating extremely comfortable and 7 indicating extremely uncomfortable). The rating distribution was as follows: 140 (67%) comfortable (rating, 1-3); 24 (12%) neither comfortable nor uncomfortable (rating, 4); and 45 (22%) uncomfortable (rating, 5 or 6). No responders gave a 7 rating. The most frequently mentioned sources of discomfort included breast compression (n = 16), back or neck discomfort (n = 14), and maintaining position during the examination (n = 14). MBI comfort was associated with responder age (74% ≥55 y old were comfortable, versus 53% <55 y old [P = 0.003]) and history of MBI (71% with prior MBI were comfortable, versus 61% having a first MBI [P = 0.006]). Of 208 responders with a prior mammogram, 148 (71%) said MBI is more comfortable than mammography (a significant majority [P < 0.001]). Of 202 responders to the question of whether they were willing to return for a future MBI, 196 (97%) were willing. A notable factor in positive patient experience was interaction with the MBI nuclear medicine technologist. Conclusion: Most responders thought MBI to be a comfortable examination and more comfortable than mammography. Patient experience during MBI may be improved by ensuring back support and soliciting patient feedback at the time of positioning and throughout the examination. Methods under study to reduce imaging time may be most important for improving patient experience.


Assuntos
Imagem Molecular , Humanos , Pessoa de Meia-Idade , Idoso , Adulto , Feminino , Inquéritos e Questionários , Idoso de 80 Anos ou mais , Imagem Molecular/métodos , Neoplasias da Mama/diagnóstico por imagem , Mamografia
11.
Molecules ; 29(11)2024 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-38893419

RESUMO

Zinc ions (Zn2+) play a key role in maintaining and regulating protein structures and functions. To better understand the intracellular Zn2+ homeostasis and signaling role, various fluorescent sensors have been developed that allow the monitoring of Zn2+ concentrations and bioimaging in live cells in real time. This review highlights the recent development of organic fluorescent probes for the detection and imaging of intracellular Zn2+, including the design and construction of the probes, fluorescent response mechanisms, and their applications to intracellular Zn2+ detection and imaging on-site. Finally, the current challenges and prospects are discussed.


Assuntos
Corantes Fluorescentes , Zinco , Corantes Fluorescentes/química , Zinco/metabolismo , Zinco/análise , Zinco/química , Humanos , Imagem Óptica/métodos , Animais , Imagem Molecular/métodos
12.
Phys Med Biol ; 69(13)2024 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-38843809

RESUMO

Objective. Image reconstruction is a fundamental step in magnetic particle imaging (MPI). One of the main challenges is the fact that the reconstructions are computationally intensive and time-consuming, so choosing an algorithm presents a compromise between accuracy and execution time, which depends on the application. This work proposes a method that provides both fast and accurate image reconstructions.Approach. Image reconstruction algorithms were implemented to be executed in parallel ingraphics processing units(GPUs) using the CUDA framework. The calculation of the model-based MPI calibration matrix was also implemented in GPU to allow both fast and flexible reconstructions.Main results. The parallel algorithms were able to accelerate the reconstructions by up to about6,100times in comparison to the serial Kaczmarz algorithm executed in the CPU, allowing for real-time applications. Reconstructions using the OpenMPIData dataset validated the proposed algorithms and demonstrated that they are able to provide both fast and accurate reconstructions. The calculation of the calibration matrix was accelerated by up to about 37 times.Significance. The parallel algorithms proposed in this work can provide single-frame MPI reconstructions in real time, with frame rates greater than 100 frames per second. The parallel calculation of the calibration matrix can be combined with the parallel reconstruction to deliver images in less time than the serial Kaczmarz reconstruction, potentially eliminating the need of storing the calibration matrix in the main memory, and providing the flexibility of redefining scanning and reconstruction parameters during execution.


Assuntos
Processamento de Imagem Assistida por Computador , Processamento de Imagem Assistida por Computador/métodos , Algoritmos , Gráficos por Computador , Fatores de Tempo , Imagem Molecular/métodos , Calibragem
14.
J Nucl Med ; 65(7): 998-1003, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38871386

RESUMO

Gynecological pathologies account for approximately 4.5% of the overall global disease burden. Although cancers of the female reproductive system have understandably been the focus of a great deal of research, benign gynecological conditions-such as endometriosis, polycystic ovary syndrome, and uterine fibroids-have remained stubbornly understudied despite their astonishing ubiquity and grave morbidity. This historical inattention has frequently become manifested in flawed diagnostic and treatment paradigms. Molecular imaging could be instrumental in improving patient care on both fronts. In this Focus on Molecular Imaging review, we will examine recent advances in the use of PET, SPECT, MRI, and fluorescence imaging for the diagnosis and management of benign gynecological conditions, with particular emphasis on recent clinical reports, areas of need, and opportunities for growth.


Assuntos
Imagem Molecular , Humanos , Imagem Molecular/métodos , Imagem Molecular/tendências , Feminino , Ginecologia
15.
Science ; 384(6701): eadh9979, 2024 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-38870291

RESUMO

Understanding cellular architectures and their connectivity is essential for interrogating system function and dysfunction. However, we lack technologies for mapping the multiscale details of individual cells and their connectivity in the human organ-scale system. We developed a platform that simultaneously extracts spatial, molecular, morphological, and connectivity information of individual cells from the same human brain. The platform includes three core elements: a vibrating microtome for ultraprecision slicing of large-scale tissues without losing cellular connectivity (MEGAtome), a polymer hydrogel-based tissue processing technology for multiplexed multiscale imaging of human organ-scale tissues (mELAST), and a computational pipeline for reconstructing three-dimensional connectivity across multiple brain slabs (UNSLICE). We applied this platform for analyzing human Alzheimer's disease pathology at multiple scales and demonstrating scalable neural connectivity mapping in the human brain.


Assuntos
Doença de Alzheimer , Encéfalo , Imagem Molecular , Humanos , Doença de Alzheimer/diagnóstico por imagem , Encéfalo/diagnóstico por imagem , Imagem Molecular/métodos , Fenótipo , Hidrogéis/química , Conectoma
16.
Anal Chem ; 96(25): 10348-10355, 2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-38857182

RESUMO

Low-field (LF) MRI promises soft-tissue imaging without the expensive, immobile magnets of clinical scanners but generally suffers from limited detection sensitivity and contrast. The sensitivity boost provided by hyperpolarization can thus be highly synergistic with LF MRI. Initial efforts to integrate a continuous-bubbling SABRE (signal amplification by reversible exchange) hyperpolarization setup with a portable, point-of-care 64 mT clinical MRI scanner are reported. Results from 1H SABRE MRI of pyrazine and nicotinamide are compared with those of benchtop NMR spectroscopy. Comparison with MRI signals from samples with known H2O/D2O ratios allowed quantification of the SABRE enhancements of imaged samples with various substrate concentrations (down to 3 mM). Respective limits of detection and quantification of 3.3 and 10.1 mM were determined with pyrazine 1H polarization (PH) enhancements of ∼1900 (PH ∼0.04%), supporting ongoing and envisioned efforts to realize SABRE-enabled MRI-based molecular imaging.


Assuntos
Imageamento por Ressonância Magnética , Imagem Molecular , Niacinamida , Sistemas Automatizados de Assistência Junto ao Leito , Pirazinas , Niacinamida/química , Imagem Molecular/métodos , Pirazinas/química , Humanos
17.
Phys Med Biol ; 69(15)2024 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-38862003

RESUMO

Objective.Magnetic particle imaging (MPI) is an emerging medical tomographic imaging modality that enables real-time imaging with high sensitivity and high spatial and temporal resolution. For the system matrix reconstruction method, the MPI reconstruction problem is an ill-posed inverse problem that is commonly solved using the Kaczmarz algorithm. However, the high computation time of the Kaczmarz algorithm, which restricts MPI reconstruction speed, has limited the development of potential clinical applications for real-time MPI. In order to achieve fast reconstruction in real-time MPI, we propose a greedy regularized block Kaczmarz method (GRBK) which accelerates MPI reconstruction.Approach.GRBK is composed of a greedy partition strategy for the system matrix, which enables preprocessing of the system matrix into well-conditioned blocks to facilitate the convergence of the block Kaczmarz algorithm, and a regularized block Kaczmarz algorithm, which enables fast and accurate MPI image reconstruction at the same time.Main results.We quantitatively evaluated our GRBK using simulation data from three phantoms at 20 dB, 30 dB, and 40 dB noise levels. The results showed that GRBK can improve reconstruction speed by single orders of magnitude compared to the prevalent regularized Kaczmarz algorithm including Tikhonov regularization, the non-negative Fused Lasso, and wavelet-based sparse model. We also evaluated our method on OpenMPIData, which is real MPI data. The results showed that our GRBK is better suited for real-time MPI reconstruction than current state-of-the-art reconstruction algorithms in terms of reconstruction speed as well as image quality.Significance.Our proposed method is expected to be the preferred choice for potential applications of real-time MPI.


Assuntos
Algoritmos , Processamento de Imagem Assistida por Computador , Imagens de Fantasmas , Processamento de Imagem Assistida por Computador/métodos , Fatores de Tempo , Tomografia/métodos , Imagem Molecular/métodos
18.
Anal Chem ; 96(24): 10084-10091, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38836421

RESUMO

Due to the potential off-tumor signal leakage and limited biomarker content, there is an urgent need for stimulus-responsive and amplification-based tumor molecular imaging strategies. Therefore, two tetrahedral framework DNA (tFNA-Hs), tFNA-H1AP, and tFNA-H2, were rationally engineered to form a polymeric tFNA network, termed an intelligent DNA network, in an AND-gated manner. The intelligent DNA network was designed for tumor-specific molecular imaging by leveraging the elevated expression of apurinic/apyrimidinic endonuclease 1 (APE1) in tumor cytoplasm instead of normal cells and the high expression of miRNA-21 in tumor cytoplasm. The activation of tFNA-H1AP can be achieved through specific recognition and cleavage by APE1, targeting the apurinic/apyrimidinic site (AP site) modified within the stem region of hairpin 1 (H1AP). Subsequently, miRNA-21 facilitates the hybridization of activated H1AP on tFNA-H1AP with hairpin 2 (H2) on tFNA-H2, triggering a catalytic hairpin assembly (CHA) reaction that opens the H1AP at the vertices of tFNA-H1AP to bind with H2 at the vertices of tFNA-H2 and generate fluorescence signals. Upon completion of hybridization, miRNA-21 is released, initiating the subsequent cycle of the CHA reaction. The AND-gated intelligent DNA network can achieve specific tumor molecular imaging in vivo and also enables risk stratification of neuroblastoma patients.


Assuntos
DNA Liase (Sítios Apurínicos ou Apirimidínicos) , DNA , MicroRNAs , Humanos , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/metabolismo , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/química , MicroRNAs/metabolismo , MicroRNAs/análise , DNA/química , DNA/metabolismo , Imagem Molecular/métodos , Animais , Imagem Óptica
19.
Methods Mol Biol ; 2822: 65-75, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38907912

RESUMO

We present a powerful method for direct mRNA detection based on ligation-based recognition and in situ amplification, capable of single-cell imaging mRNA at single-nucleotide and single-molecule resolution. Attributed to the use of Splint R ligase that can ligate padlock probe with RNA as target template, this method can efficiently detect mRNA in the absence of reverse transcription. This method enables spatial localization and correlation analysis of gene expression in single cells, which helps us to elucidate gene function and regulatory mechanisms.


Assuntos
RNA Mensageiro , Análise de Célula Única , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Análise de Célula Única/métodos , Humanos , Técnicas de Amplificação de Ácido Nucleico/métodos , Imagem Individual de Molécula/métodos , Imagem Molecular/métodos
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