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1.
Br J Cancer ; 131(3): 457-467, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38902534

RESUMEN

BACKGROUND/OBJECTIVES: Pseudo-vascular network formation in vitro is considered a key characteristic of vasculogenic mimicry. While many cancer cell lines form pseudo-vascular networks, little is known about the spatiotemporal dynamics of these formations. METHODS: Here, we present a framework for monitoring and characterising the dynamic formation and dissolution of pseudo-vascular networks in vitro. The framework combines time-resolved optical microscopy with open-source image analysis for network feature extraction and statistical modelling. The framework is demonstrated by comparing diverse cancer cell lines associated with vasculogenic mimicry, then in detecting response to drug compounds proposed to affect formation of vasculogenic mimics. Dynamic datasets collected were analysed morphometrically and a descriptive statistical analysis model was developed in order to measure stability and dissimilarity characteristics of the pseudo-vascular networks formed. RESULTS: Melanoma cells formed the most stable pseudo-vascular networks and were selected to evaluate the response of their pseudo-vascular networks to treatment with axitinib, brucine and tivantinib. Tivantinib has been found to inhibit the formation of the pseudo-vascular networks more effectively, even in dose an order of magnitude less than the two other agents. CONCLUSIONS: Our framework is shown to enable quantitative analysis of both the capacity for network formation, linked vasculogenic mimicry, as well as dynamic responses to treatment.


Asunto(s)
Neovascularización Patológica , Humanos , Neovascularización Patológica/tratamiento farmacológico , Neovascularización Patológica/patología , Línea Celular Tumoral , Melanoma/patología , Melanoma/irrigación sanguínea , Melanoma/tratamiento farmacológico , Axitinib/farmacología
2.
Angew Chem Int Ed Engl ; 63(25): e202404885, 2024 06 17.
Artículo en Inglés | MEDLINE | ID: mdl-38622059

RESUMEN

There is an urgent need to improve conventional cancer-treatments by preventing detrimental side effects, cancer recurrence and metastases. Recent studies have shown that presence of senescent cells in tissues treated with chemo- or radiotherapy can be used to predict the effectiveness of cancer treatment. However, although the accumulation of senescent cells is one of the hallmarks of cancer, surprisingly little progress has been made in development of strategies for their detection in vivo. To address a lack of detection tools, we developed a biocompatible, injectable organic nanoprobe (NanoJagg), which is selectively taken up by senescent cells and accumulates in the lysosomes. The NanoJagg probe is obtained by self-assembly of indocyanine green (ICG) dimers using a scalable manufacturing process and characterized by a unique spectral signature suitable for both photoacoustic tomography (PAT) and fluorescence imaging. In vitro, ex vivo and in vivo studies all indicate that NanoJaggs are a clinically translatable probe for detection of senescence and their PAT signal makes them suitable for longitudinal monitoring of the senescence burden in solid tumors after chemotherapy or radiotherapy.


Asunto(s)
Senescencia Celular , Verde de Indocianina , Verde de Indocianina/química , Senescencia Celular/efectos de los fármacos , Humanos , Animales , Imagen Óptica , Ratones , Nanopartículas/química , Colorantes Fluorescentes/química , Técnicas Fotoacústicas/métodos
3.
Opt Express ; 30(5): 7591-7611, 2022 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-35299518

RESUMEN

Multispectral imaging captures spatial information across a set of discrete spectral channels and is widely utilized across diverse applications such as remote sensing, industrial inspection, and biomedical imaging. Multispectral filter arrays (MSFAs) are filter mosaics integrated atop image sensors that facilitate cost-effective, compact, snapshot multispectral imaging. MSFAs are pre-configured based on application-where filter channels are selected corresponding to targeted absorption spectra-making the design of optimal MSFAs vital for a given application. Despite the availability of many design and optimization approaches for spectral channel selection and spatial arrangement, major limitations remain. There are few robust approaches for joint spectral-spatial optimization, techniques are typically only applicable to limited datasets and most critically, are not available for general use and improvement by the wider community. Here, we reconcile current MSFA design techniques and present Opti-MSFA: a Python-based open-access toolbox for the centralized design and optimization of MSFAs. Opti-MSFA incorporates established spectral-spatial optimization algorithms, such as gradient descent and simulated annealing, multispectral-RGB image reconstruction, and is applicable to user-defined input of spatial-spectral datasets or imagery. We demonstrate the utility of the toolbox by comparing against other published MSFAs using the standard hyperspectral datasets Samson and Jasper Ridge, and further show application on experimentally acquired fluorescence imaging data. In conjunction with end-user input and collaboration, we foresee the continued development of Opti-MSFA for the benefit of the wider research community.

4.
Breast Cancer Res ; 23(1): 25, 2021 02 17.
Artículo en Inglés | MEDLINE | ID: mdl-33596961

RESUMEN

BACKGROUND: Ductal carcinoma in situ (DCIS) is a non-invasive form of early breast cancer, with a poorly understood natural history of invasive transformation. Necrosis is a well-recognized adverse prognostic feature of DCIS, and non-invasive detection of its presence and spatial extent could provide information not obtainable by biopsy. We describe here imaging of the distribution and extent of comedo-type necrosis in a model of human DCIS using C2Am, an imaging agent that binds to the phosphatidylserine exposed by necrotic cells. METHODS: We used an established xenograft model of human DCIS that mimics the histopathological features of the disease. Planar near-infrared and optoacoustic imaging, using fluorescently labeled C2Am, were used to image non-invasively the presence and extent of lesion necrosis. RESULTS: C2Am showed specific and sensitive binding to necrotic areas in DCIS tissue, detectable both in vivo and ex vivo. The imaging signal generated in vivo using near-infrared (NIR) fluorescence imaging was up to 6-fold higher in DCIS lesions than in surrounding fat pad or skin tissue. There was a correlation between the C2Am NIR fluorescence (Pearson R = 0.783, P = 0.0125) and optoacoustic signals (R > 0.875, P < 0.022) in the DCIS lesions in vivo and the corresponding levels of cell death detected histologically. CONCLUSIONS: C2Am is a targeted multi-modal imaging agent that could complement current anatomical imaging methods for detecting DCIS. Imaging the presence and spatial extent of necrosis may give better prognostic information than that obtained by biopsy alone.


Asunto(s)
Neoplasias de la Mama/diagnóstico por imagen , Neoplasias de la Mama/patología , Carcinoma in Situ/diagnóstico por imagen , Carcinoma in Situ/patología , Carcinoma Ductal de Mama/diagnóstico por imagen , Carcinoma Ductal de Mama/patología , Imagen Multimodal , Animales , Muerte Celular , Línea Celular Tumoral , Medios de Contraste , Modelos Animales de Enfermedad , Detección Precoz del Cáncer , Femenino , Humanos , Inmunohistoquímica , Ratones , Imagen Molecular , Imagen Multimodal/métodos , Imagen Multimodal/normas , Imagen Óptica , Técnicas Fotoacústicas
5.
Nat Methods ; 13(8): 639-50, 2016 07 28.
Artículo en Inglés | MEDLINE | ID: mdl-27467727

RESUMEN

Photoacoustic imaging (PAI) is an emerging tool that bridges the traditional depth limits of ballistic optical imaging and the resolution limits of diffuse optical imaging. Using the acoustic waves generated in response to the absorption of pulsed laser light, it provides noninvasive images of absorbed optical energy density at depths of several centimeters with a resolution of ∼100 µm. This versatile and scalable imaging modality has now shown potential for molecular imaging, which enables visualization of biological processes with systemically introduced contrast agents. Understanding the relative merits of the vast range of contrast agents available, from small-molecule dyes to gold and carbon nanostructures to liposome encapsulations, is a considerable challenge. Here we critically review the physical, chemical and biochemical characteristics of the existing photoacoustic contrast agents, highlighting key applications and present challenges for molecular PAI.


Asunto(s)
Medios de Contraste/química , Diagnóstico por Imagen/métodos , Técnicas Fotoacústicas/métodos , Imagen de Cuerpo Entero/métodos , Animales , Humanos , Nanopartículas/química
6.
Opt Express ; 27(17): 23929-23947, 2019 Aug 19.
Artículo en Inglés | MEDLINE | ID: mdl-31510290

RESUMEN

Flexible optical fibres, used in conventional medical endoscopy and industrial inspection, scramble phase and polarisation information, restricting users to amplitude-only imaging. Here, we exploit the near-diagonality of the multi-core fibre (MCF) transmission matrix in a parallelised fibre characterisation architecture, enabling accurate imaging of quantitative phase (error <0.3 rad) and polarisation-resolved (errors <10%) properties. We first demonstrate accurate recovery of optical amplitude and phase in two polarisations through the MCF by measuring and inverting the transmission matrix, and then present a robust Bayesian inference approach to resolving 5 polarimetric properties of samples. Our method produces high-resolution (9.0±2.6µm amplitude, phase; 36.0±10.4µm polarimetric) full-field images at working distances up to 1mm over a field-of-view up to 750×750µm 2 using an MCF with potential for flexible operation. We demonstrate the potential of using quantitative phase for computational image focusing and polarisation-resolved properties in imaging birefringence.

7.
J Biol Chem ; 292(5): 1737-1748, 2017 02 03.
Artículo en Inglés | MEDLINE | ID: mdl-27994059

RESUMEN

Rapid cancer cell proliferation promotes the production of reducing equivalents, which counteract the effects of relatively high levels of reactive oxygen species. Reactive oxygen species levels increase in response to chemotherapy and cell death, whereas an increase in antioxidant capacity can confer resistance to chemotherapy and is associated with an aggressive tumor phenotype. The pentose phosphate pathway is a major site of NADPH production in the cell, which is used to maintain the main intracellular antioxidant, glutathione, in its reduced state. Previous studies have shown that the rate of hyperpolarized [1-13C]dehydroascorbic acid (DHA) reduction, which can be measured in vivo using non-invasive 13C magnetic resonance spectroscopic imaging, is increased in tumors and that this is correlated with the levels of reduced glutathione. We show here that the rate of hyperpolarized [1-13C]DHA reduction is increased in tumors that have been oxidatively prestressed by depleting the glutathione pool by buthionine sulfoximine treatment. This increase was associated with a corresponding increase in pentose phosphate pathway flux, assessed using 13C-labeled glucose, and an increase in glutaredoxin activity, which catalyzes the glutathione-dependent reduction of DHA. These results show that the rate of DHA reduction depends not only on the level of reduced glutathione, but also on the rate of NADPH production, contradicting the conclusions of some previous studies. Hyperpolarized [1-13C]DHA can be used, therefore, to assess the capacity of tumor cells to resist oxidative stress in vivo However, DHA administration resulted in transient respiratory arrest and cardiac depression, which may prevent translation to the clinic.


Asunto(s)
Ácido Deshidroascórbico/metabolismo , NADP/metabolismo , Neoplasias/metabolismo , Estrés Oxidativo , Animales , Isótopos de Carbono , Línea Celular Tumoral , Humanos , Marcaje Isotópico , Espectroscopía de Resonancia Magnética , Ratones
8.
Br J Cancer ; 118(8): 1098-1106, 2018 04.
Artículo en Inglés | MEDLINE | ID: mdl-29576623

RESUMEN

BACKGROUND: Optoacoustic tomography (OT) of breast tumour oxygenation is a promising new technique, currently in clinical trials, which may help to determine disease stage and therapeutic response. However, the ability of OT to distinguish breast tumours displaying different vascular characteristics has yet to be established. The aim of the study is to prove OT as a sensitive technique for differentiating breast tumour models with manifestly different vasculatures. METHODS: Multispectral OT (MSOT) was performed in oestrogen-dependent (MCF-7) and oestrogen-independent (MDA-MB-231) orthotopic breast cancer xenografts. Total haemoglobin (THb) and oxygen saturation (SO2MSOT) were calculated. Pathological and biochemical evaluation of the tumour vascular phenotype was performed for validation. RESULTS: MCF-7 tumours show SO2MSOT similar to healthy tissue in both rim and core, despite significantly lower THb in the core. MDA-MB-231 tumours show markedly lower SO2MSOT with a significant rim-core disparity. Ex vivo analysis revealed that MCF-7 tumours contain fewer blood vessels (CD31+) that are more mature (CD31+/aSMA+) than MDA-MB-231. MCF-7 presented higher levels of stromal VEGF and iNOS, with increased NO serum levels. The vasculogenic process observed in MCF-7 was consistent with angiogenesis, while MDA-MB-231 appeared to rely more on vascular mimicry. CONCLUSIONS: OT is sensitive to differences in the vascular phenotypes of our breast cancer models.


Asunto(s)
Mimetismo Biológico/fisiología , Neoplasias Mamarias Experimentales/irrigación sanguínea , Neoplasias Mamarias Experimentales/diagnóstico , Neoplasias Mamarias Experimentales/patología , Neovascularización Patológica/diagnóstico , Técnicas Fotoacústicas/métodos , Tomografía/métodos , Animales , Neoplasias de la Mama/irrigación sanguínea , Neoplasias de la Mama/diagnóstico , Neoplasias de la Mama/patología , Línea Celular Tumoral , Monitoreo de Drogas/métodos , Femenino , Humanos , Células MCF-7 , Neoplasias Mamarias Experimentales/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Estadificación de Neoplasias , Neovascularización Patológica/patología , Consumo de Oxígeno/fisiología , Sensibilidad y Especificidad , Hipoxia Tumoral/fisiología , Ensayos Antitumor por Modelo de Xenoinjerto
9.
Small ; 14(19): e1703683, 2018 05.
Artículo en Inglés | MEDLINE | ID: mdl-29635739

RESUMEN

Raman microspectroscopy provides chemo-selective image contrast, sub-micrometer resolution, and multiplexing capabilities. However, it suffers from weak signals resulting in image-acquisition times of up to several hours. Surface-enhanced Raman scattering (SERS) can dramatically enhance signals of molecules in close vicinity of metallic surfaces and overcome this limitation. Multimodal, SERS-active nanoparticles are usually labeled with Raman marker molecules, limiting SERS to the coating material. In order to realize multimodal imaging while acquiring the rich endogenous vibronic information of the specimen, a core-shell particle based on "Nanorice", where a spindle-shaped iron oxide core is encapsulated by a closed gold shell, is developed. An ultrathin layer of silica prevents agglomeration and unwanted chemical interaction with the specimen. This approach provides Raman signal enhancement due to plasmon resonance effects of the shell while the optical absorption in the near-infrared spectral region provides contrast in photoacoustic tomography. Finally, T2-relaxation of a magnetic resonance imaging (MRI) experiment is altered by taking advantage of the iron oxide core. The feasibility for Raman imaging is evaluated by nearfield simulations and experimental studies on the primate cell line COS1. MRI and photoacoustics are demonstrated in agarose phantoms illustrating the promising translational nature of this strategy for clinical applications in radiology.


Asunto(s)
Medios de Contraste/química , Polvo , Imagen por Resonancia Magnética/métodos , Nanopartículas/química , Técnicas Fotoacústicas/métodos , Espectrometría Raman , Animales , Células COS , Chlorocebus aethiops , Simulación por Computador , Nanopartículas/ultraestructura , Fantasmas de Imagen
10.
Endoscopy ; 50(6): 618-625, 2018 06.
Artículo en Inglés | MEDLINE | ID: mdl-29342490

RESUMEN

BACKGROUND AND STUDY AIMS: Endoscopic surveillance for Barrett's esophagus (BE) is limited by long procedure times and sampling error. Near-infrared (NIR) fluorescence imaging minimizes tissue autofluorescence and optical scattering. We assessed the feasibility of a topically applied NIR dye-labeled lectin for the detection of early neoplasia in BE in an ex vivo setting. METHODS: Consecutive patients undergoing endoscopic mucosal resection (EMR) for BE-related early neoplasia were recruited. Freshly collected EMR specimens were sprayed at the bedside with fluorescent lectin and then imaged. Punch biopsies were collected from each EMR under NIR light guidance. We compared the fluorescence intensity from dysplastic and nondysplastic areas within EMRs and from punch biopsies with different histological grades. RESULTS: 29 EMR specimens were included from 17 patients. A significantly lower fluorescence was found for dysplastic regions across whole EMR specimens (P < 0.001). We found a 41 % reduction in the fluorescence of dysplastic compared to nondysplastic punch biopsies (P < 0.001), with a sensitivity and specificity for dysplasia detection of 80 % and 82.9 %, respectively. CONCLUSION: Lectin-based NIR imaging can differentiate dysplastic from nondysplastic Barrett's mucosa ex vivo.


Asunto(s)
Esófago de Barrett/diagnóstico por imagen , Neoplasias Esofágicas/diagnóstico por imagen , Esófago/patología , Lectinas/análisis , Imagen Molecular/métodos , Imagen Óptica/métodos , Anciano , Anciano de 80 o más Años , Esófago de Barrett/patología , Esófago de Barrett/cirugía , Biopsia , Resección Endoscópica de la Mucosa , Neoplasias Esofágicas/patología , Neoplasias Esofágicas/cirugía , Estudios de Factibilidad , Femenino , Fluorescencia , Humanos , Hiperplasia/diagnóstico por imagen , Hiperplasia/patología , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Coloración y Etiquetado
11.
Appl Opt ; 56(12): 3303-3310, 2017 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-28430250

RESUMEN

Optical fiber technology is found in a wide variety of applications to flexibly relay light between two points, enabling information transfer across long distances and allowing access to hard-to-reach areas. Large-core optical fibers and light guides find frequent use in illumination and spectroscopic applications, for example, endoscopy and high-resolution astronomical spectroscopy. Proper alignment is critical for maximizing throughput in optical fiber coupling systems; however, there currently are no formal approaches to tolerancing the alignment of a light-guide coupling system. Here, we propose a Fourier alignment sensitivity (FAS) algorithm to determine the optimal tolerances on the alignment of a light guide by computing the alignment sensitivity. The algorithm shows excellent agreement with both simulated and experimentally measured values and improves on the computation time of equivalent ray-tracing simulations by two orders of magnitude. We then apply FAS to tolerance and fabricate a coupling system, which is shown to meet specifications, thus validating FAS as a tolerancing technique. These results indicate that FAS is a flexible and rapid means to quantify the alignment sensitivity of a light guide, widely informing the design and tolerancing of coupling systems.

13.
Proc Natl Acad Sci U S A ; 110(30): 12408-13, 2013 Jul 23.
Artículo en Inglés | MEDLINE | ID: mdl-23821752

RESUMEN

Raman spectroscopy, amplified by surface enhanced Raman scattering (SERS) nanoparticles, is a molecular imaging modality with ultra-high sensitivity and the unique ability to multiplex readouts from different molecular targets using a single wavelength of excitation. This approach holds exciting prospects for a range of applications in medicine, including identification and characterization of malignancy during endoscopy and intraoperative image guidance of surgical resection. The development of Raman molecular imaging with SERS nanoparticles is presently limited by long acquisition times, poor spatial resolution, small field of view, and difficulty in animal handling with existing Raman spectroscopy instruments. Our goal is to overcome these limitations by designing a bespoke instrument for Raman molecular imaging in small animals. Here, we present a unique and dedicated small-animal Raman imaging instrument that enables rapid, high-spatial resolution, spectroscopic imaging over a wide field of view (> 6 cm(2)), with simplified animal handling. Imaging of SERS nanoparticles in small animals demonstrated that this small animal Raman imaging system can detect multiplexed SERS signals in both superficial and deep tissue locations at least an order of magnitude faster than existing systems without compromising sensitivity.


Asunto(s)
Espectrometría Raman/métodos , Animales , Femenino , Ratones , Ratones Desnudos
14.
J Lightwave Technol ; 33(16): 3419-3425, 2015 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-27279676

RESUMEN

In this paper a novel single-pixel method for coherent imaging through an endoscopic fiber bundle is presented. The use of a single-pixel detector allows greater sensitivity over a wider range of wavelengths, which could have significant applications in endoscopic fluorescence microscopy. First, the principle of lensless focussing at the distal end of a coherent fiber bundle is simulated to examine the impact of pixelation at microscopic scales. Next, an experimental optical correlator system using spatial light modulators (SLMs) is presented. A simple contrast imaging method of characterizing and compensating phase aberrations introduced by fiber bundles is described. Experimental results are then presented showing that our phase compensation method enables characterization of the optical phase profile of individual fiberlets. After applying this correction, early results demonstrating the ability of the system to electronically adjust the focal plane at the distal end of the fiber bundle are presented. The structural similarity index (SSIM) between the simulated image and the experimental focus-adjusted image increases noticeably when the phase correction is applied and the retrieved image is visually recognizable. Strategies to improve image quality are discussed.

15.
Proc Natl Acad Sci U S A ; 109(33): 13374-9, 2012 Aug 14.
Artículo en Inglés | MEDLINE | ID: mdl-22837393

RESUMEN

Acute kidney injury (AKI) is a common and important medical problem, affecting 10% of hospitalized patients, and it is associated with significant morbidity and mortality. The most frequent cause of AKI is acute tubular necrosis (ATN). Current imaging techniques and biomarkers do not allow ATN to be reliably differentiated from important differential diagnoses, such as acute glomerulonephritis (GN). We investigated whether (13)C magnetic resonance spectroscopic imaging (MRSI) might allow the noninvasive diagnosis of ATN. (13)C MRSI of hyperpolarized [1,4-(13)C(2)]fumarate and pyruvate was used in murine models of ATN and acute GN (NZM2410 mice with lupus nephritis). A significant increase in [1,4-(13)C(2)]malate signal was identified in the kidneys of mice with ATN early in the disease course before the onset of severe histological changes. No such increase in renal [1,4-(13)C(2)]malate was observed in mice with acute GN. The kidney [1-(13)C]pyruvate/[1-(13)C]lactate ratio showed substantial variability and was not significantly decreased in animals with ATN or increased in animals with GN. In conclusion, MRSI of hyperpolarized [1,4-(13)C(2)]fumarate allows the detection of early tubular necrosis and its distinction from glomerular inflammation in murine models. This technique may have the potential to identify a window of therapeutic opportunity in which emerging therapies might be applied to patients with ATN, reducing the need for acute dialysis with its attendant morbidity and cost.


Asunto(s)
Fumaratos , Necrosis Tubular Aguda/diagnóstico , Imagen por Resonancia Magnética/métodos , Animales , Isótopos de Carbono , Diagnóstico Precoz , Ácido Fólico , Humanos , Riñón/anomalías , Riñón/patología , Riñón/fisiopatología , Necrosis Tubular Aguda/inducido químicamente , Necrosis Tubular Aguda/fisiopatología , Cinética , Nefritis Lúpica/diagnóstico , Nefritis Lúpica/patología , Malatos , Ratones , Ratones Endogámicos C57BL , Ácido Pirúvico
16.
Magn Reson Med ; 71(1): 402-10, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-23440731

RESUMEN

PURPOSE: The acquisition of ever increasing volumes of high resolution magnetic resonance imaging (MRI) data has created an urgent need to develop automated and objective image analysis algorithms that can assist in determining tumor margins, diagnosing tumor stage, and detecting treatment response. METHODS: We have shown previously that Minkowski functionals, which are precise morphological and structural descriptors of image heterogeneity, can be used to enhance the detection, in T1 -weighted images, of a targeted Gd(3+) -chelate-based contrast agent for detecting tumor cell death. We have used Minkowski functionals here to characterize heterogeneity in T2 -weighted images acquired before and after drug treatment, and obtained without contrast agent administration. RESULTS: We show that Minkowski functionals can be used to characterize the changes in image heterogeneity that accompany treatment of tumors with a vascular disrupting agent, combretastatin A4-phosphate, and with a cytotoxic drug, etoposide. CONCLUSIONS: Parameterizing changes in the heterogeneity of T2 -weighted images can be used to detect early responses of tumors to drug treatment, even when there is no change in tumor size. The approach provides a quantitative and therefore objective assessment of treatment response that could be used with other types of MR image and also with other imaging modalities.


Asunto(s)
Etopósido/uso terapéutico , Interpretación de Imagen Asistida por Computador/métodos , Linfoma/tratamiento farmacológico , Linfoma/patología , Imagen por Resonancia Magnética/métodos , Estilbenos/uso terapéutico , Animales , Antineoplásicos/uso terapéutico , Línea Celular Tumoral , Femenino , Ratones , Ratones Endogámicos C57BL , Estadificación de Neoplasias , Pronóstico , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Resultado del Tratamiento
17.
Nat Commun ; 15(1): 4135, 2024 May 16.
Artículo en Inglés | MEDLINE | ID: mdl-38755150

RESUMEN

Point-scanning microscopy approaches are transforming super-resolution imaging. Despite achieving parallel high-speed imaging using multifocal techniques, efficient multicolor imaging methods with high-quality illumination are currently lacking. In this paper, we present for the first time Mechanical-scan-free multiColor Super-resolution Microscopy (MCoSM) with spot array illumination, which enables mechanical-scan-free super-resolution imaging with adjustable resolution and a good effective field-of-view based on spatial light modulators. Through 100-2,500 s super-resolution spot illumination with different effective fields of view for imaging, we demonstrate the adjustable capacity of MCoSM. MCoSM extends existing spectral imaging capabilities through a time-sharing process involving different color illumination with phase-shift scanning while retaining the spatial flexibility of super-resolution imaging with diffractive spot array illumination. To demonstrate the prospects of MCoSM, we perform four-color imaging of fluorescent beads at high resolution. MCoSM provides a versatile platform for studying molecular interactions in complex samples at the nanoscale level.

18.
IEEE Trans Med Imaging ; 43(3): 1214-1224, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-37938947

RESUMEN

Accurate measurement of optical absorption coefficients from photoacoustic imaging (PAI) data would enable direct mapping of molecular concentrations, providing vital clinical insight. The ill-posed nature of the problem of absorption coefficient recovery has prohibited PAI from achieving this goal in living systems due to the domain gap between simulation and experiment. To bridge this gap, we introduce a collection of experimentally well-characterised imaging phantoms and their digital twins. This first-of-a-kind phantom data set enables supervised training of a U-Net on experimental data for pixel-wise estimation of absorption coefficients. We show that training on simulated data results in artefacts and biases in the estimates, reinforcing the existence of a domain gap between simulation and experiment. Training on experimentally acquired data, however, yielded more accurate and robust estimates of optical absorption coefficients. We compare the results to fluence correction with a Monte Carlo model from reference optical properties of the materials, which yields a quantification error of approximately 20%. Application of the trained U-Nets to a blood flow phantom demonstrated spectral biases when training on simulated data, while application to a mouse model highlighted the ability of both learning-based approaches to recover the depth-dependent loss of signal intensity. We demonstrate that training on experimental phantoms can restore the correlation of signal amplitudes measured in depth. While the absolute quantification error remains high and further improvements are needed, our results highlight the promise of deep learning to advance quantitative PAI.


Asunto(s)
Técnicas Fotoacústicas , Animales , Ratones , Fantasmas de Imagen , Técnicas Fotoacústicas/métodos , Diagnóstico por Imagen , Simulación por Computador , Método de Montecarlo
19.
J Biomed Opt ; 29(3): 036005, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38560531

RESUMEN

Significance: Color differences between healthy and diseased tissue in the gastrointestinal (GI) tract are detected visually by clinicians during white light endoscopy; however, the earliest signs of cancer are often just a slightly different shade of pink compared to healthy tissue making it hard to detect. Improving contrast in endoscopy is important for early detection of disease in the GI tract during routine screening and surveillance. Aim: We aim to target alternative colors for imaging to improve contrast using custom multispectral filter arrays (MSFAs) that could be deployed in an endoscopic "chip-on-tip" configuration. Approach: Using an open-source toolbox, Opti-MSFA, we examined the optimal design of MSFAs for early cancer detection in the GI tract. The toolbox was first extended to use additional classification models (k-nearest neighbor, support vector machine, and spectral angle mapper). Using input spectral data from published clinical trials examining the esophagus and colon, we optimized the design of MSFAs with three to nine different bands. Results: We examined the variation of the spectral and spatial classification accuracies as a function of the number of bands. The MSFA configurations tested showed good classification accuracies when compared to the full hyperspectral data available from the clinical spectra used in these studies. Conclusion: The ability to retain good classification accuracies with a reduced number of spectral bands could enable the future deployment of multispectral imaging in an endoscopic chip-on-tip configuration using simplified MSFA hardware. Further studies using an expanded clinical dataset are needed to confirm these findings.


Asunto(s)
Endoscopía Gastrointestinal , Neoplasias , Humanos , Diagnóstico por Imagen , Esófago
20.
J Biomed Opt ; 29(Suppl 1): S11506, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38125716

RESUMEN

Significance: Photoacoustic imaging (PAI) provides contrast based on the concentration of optical absorbers in tissue, enabling the assessment of functional physiological parameters such as blood oxygen saturation (sO2). Recent evidence suggests that variation in melanin levels in the epidermis leads to measurement biases in optical technologies, which could potentially limit the application of these biomarkers in diverse populations. Aim: To examine the effects of skin melanin pigmentation on PAI and oximetry. Approach: We evaluated the effects of skin tone in PAI using a computational skin model, two-layer melanin-containing tissue-mimicking phantoms, and mice of a consistent genetic background with varying pigmentations. The computational skin model was validated by simulating the diffuse reflectance spectrum using the adding-doubling method, allowing us to assign our simulation parameters to approximate Fitzpatrick skin types. Monte Carlo simulations and acoustic simulations were run to obtain idealized photoacoustic images of our skin model. Photoacoustic images of the phantoms and mice were acquired using a commercial instrument. Reconstructed images were processed with linear spectral unmixing to estimate blood oxygenation. Linear unmixing results were compared with a learned unmixing approach based on gradient-boosted regression. Results: Our computational skin model was consistent with representative literature for in vivo skin reflectance measurements. We observed consistent spectral coloring effects across all model systems, with an overestimation of sO2 and more image artifacts observed with increasing melanin concentration. The learned unmixing approach reduced the measurement bias, but predictions made at lower blood sO2 still suffered from a skin tone-dependent effect. Conclusion: PAI demonstrates measurement bias, including an overestimation of blood sO2, in higher Fitzpatrick skin types. Future research should aim to characterize this effect in humans to ensure equitable application of the technology.


Asunto(s)
Técnicas Fotoacústicas , Pigmentación de la Piel , Humanos , Animales , Ratones , Oxígeno , Melaninas , Técnicas Fotoacústicas/métodos , Oximetría/métodos , Fantasmas de Imagen
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