I13-2-Diamond Manchester Imaging
|
Sissi
Dolci
,
Loris
Mannino
,
Eros
Rossi
,
Emanuela
Bottani
,
Francesca
Ciarpella
,
Nicola
Piazza
,
Isabel
Karkossa
,
Marzia
Di Chio
,
Benedetta
Savino
,
Benedetta
Lucidi
,
Giulia
Pruonto
,
Ilaria
Barone
,
Alessandra
Campanelli
,
Francesca
Cersosimo
,
Elisa
Setten
,
Stefano
Gianoli
,
Zulkifal
Malik
,
Giuseppe
Busetto
,
Alex
Pezzotta
,
Alessandra
Castagna
,
Nicolò
Martinelli
,
Silvia
Ferretti
,
Federico
Boschi
,
Adam
Doherty
,
Maria Teresa
Scupoli
,
Chiara
Cavallini
,
Giorgio
Malpeli
,
Alessia
Amenta
,
Ludovica
Sagripanti
,
Vincenzo
Silani
,
Patrizia
Cristofori
,
Eugenio
Scanziani
,
Marco
Sandri
,
Anna
Pistocchi
,
Patrizia
Bossolasco
,
Marco
Endrizzi
,
Kristin
Schubert
,
Guido Francesco
Fumagalli
,
Massimo
Locati
,
Francesco
Bifari
,
Ilaria
Decimo
Diamond Proposal Number(s):
[32416]
Open Access
Abstract: Tumor-associated macrophages (TAMs) enhance cancer progression by promoting angiogenesis, extracellular matrix remodeling, and immune suppression. Nerve infiltration also contributes to tumor growth. However, the role of TAMs in promoting intratumoral nerve growth remains unclear. In this study, we have shown that TAMs express a distinct neural growth gene signature. TAMs actively enhanced neural growth within tumors and directly promoted in vitro neurite outgrowth. We identified secreted phosphoprotein 1 (SPP1) as a required mediator of TAM-driven neural growth and mTORC2 activation. Leveraging this TAM-neural growth function, we explored TAM neuroregenerative potential. Adoptive transfer of TAMs in severe complete-compressive-contusive spinal cord injury (scSCI) increased neuronal survival, axonal regrowth, and motor function recovery. Moreover, TAMs healed scSCI microenvironment and remodeled the cyst. Functional and proteomic analyses confirmed SPP1 and neural Rictor as necessary molecular mediators for TAM-induced regeneration. Our data unveil a role for TAMs in tumor innervation and neural tissue repair.
|
Jan 2026
|
|
I13-2-Diamond Manchester Imaging
|
Josh
Williams
,
Rudolf
Hellmuth
,
Yuan-Tsan
Tseng
,
Marta
Pena Fernandez
,
Oriol
Roche I Morgo
,
Yunpeng
Jia
,
Marco
Endrizzi
,
Kazimir
Wenelik
,
Leonard
Turpin
,
Shashidhara
Marathe
,
Magdi
Yacoub
Abstract: Micro-computational tomography (µCT) is a useful technique for acquiring 3-D imaging of tissue-engineered scaffolds for morphology characterisation and analysis of the mechanical interactions between scaffold and cells. We used synchrotron light µCT at Diamond Light Source (UK) to image jet-sprayed nonwoven fibrous scaffolds, with and without human adipose-derived stem cells.
Large-volume imaging was achieved by stitching 2×2 tiled datasets and reconstructing them into 1 mm³ volumes at sub-micron resolution, enabling clear scaffold segmentation from the background. However, cells and fibres produce the same X-ray attenuation, this provides challenges in segmentation between fibres and cells. A deep learning algorithm with morphological recognition was employed. It enabled rapid selective segmentation, which allowed the analysis of cell distribution and morphology, revealing that cells preferentially adhered and proliferated along in-plane structures at full scaffold colonisation. We hypothesise that the cells minimise energy expenditure by expanding in directions of least resistance.
This process for analysing tissue-engineered scaffold opens new avenues for rapid, non-destructive, high-resolution, large-volume characterisation to elucidate cell and structural interaction.
|
Sep 2025
|
|
I13-2-Diamond Manchester Imaging
|
Abstract: Cell behaviour and tissue development are inherently sensitive to morphological features of tissue-engineered scaffolds. Traditionally, imaging techniques such as SEM, TEM, AFM, and CLSM provide high-resolution 2D images to characterise scaffold morphology. However, these techniques have poor penetration and low resolution transversely to the sliced planes. In contrast, synchrotron radiation X-ray micro-computed tomography (SR-µCT) enables 3-D imaging of large volumes with submicron isotropic resolution.
We used SR-µCT at beamline I13-2 (Diamond Light Source) to image jet-sprayed nonwoven fibrous scaffolds used in the Harefield Valve, both with and without human adipose-derived stem cells preserved in ethanol to maintain native wet conditions. Large-volume imaging was achieved by stitching 2x2 tiled datasets and reconstructing them into 1 mm³ volumes at 0.325 µm voxel size, enabling clear scaffold.
The scaffold exhibited a layered, transversely isotropic structure, with additional in-plane anisotropy observed when using high-speed drum fabrication. SR-µCT revealed significantly higher scaffold porosity compared to SEM analysis, which consistently underestimates porosity due to limited depth and connectivity information. Cell distribution and morphology showed that cells preferentially adhered and proliferated along in-plane structures at full scaffold colonisation. We hypothesise that the cells minimise energy expenditure by expanding in directions of least resistance.
|
Sep 2025
|
|
I13-2-Diamond Manchester Imaging
|
Diamond Proposal Number(s):
[17971]
Open Access
Abstract: We propose an imaging system and methodology for mapping soft-tissue samples in three dimensions, with micron-scale and isotropic spatial resolution, with low-concentrations as well as in the absence of heavy metal staining. We used hard x-ray phase-contrast imaging for the x-ray ability to nondestructively probe the internal structure of opaque specimens and for enhanced contrast obtained by exploiting phase effects, even in cases with reduced or absent staining agents. To demonstrate its applicability to soft-tissue specimens, we built a compact system that is easily deployable in a laboratory setting. The imaging system is based on a conventional rotating anode x-ray tube and a state-of-the-art custom-made radiation detector. The systems performance is quantitatively assessed on a calibration standard. Its potential for soft-tissue microscopy is demonstrated on two biological specimens and benchmarked against gold-standard synchrotron data. We believe that the approach proposed here can be valuable as a bridging imaging modality for intravital correlative light and electron microscopy and be applied across disciplines where the three-dimensional morphology of pristine-condition soft tissues is a key element of the investigation.
|
Jan 2025
|
|
I13-2-Diamond Manchester Imaging
|
Diamond Proposal Number(s):
[30748]
Open Access
Abstract: The poor soft tissue contrast of X-ray CT necessitates contrast agent use to improve diagnosis across disease applications, yet their poor detection sensitivity requires high injected doses, which restrict use in at-risk populations. Dark-field X-ray imaging is emerging as a more sensitive alternative to traditional attenuation-based imaging, leveraging scattered radiation to produce contrast. Yet aside from large, short-lived microbubbles, the alternate physics of dark-field detection has yet to be exploited for contrast agent development. Here we demonstrate that high-Z nanoparticles can provide a new means to producing dark-field image contrast, promoting scatter via a higher rather than lower electron density compared to microbubbles, increasing detection sensitivity compared to attenuation-based detection of a clinical iodine-based agent at an equivalent X-ray dose. As the use of dark-field X-ray imaging expands into more common clinical usage, this will support the development of a new class of nanoparticulate contrast agents.
|
Nov 2024
|
|
I13-2-Diamond Manchester Imaging
|
Carlos
Navarrete-Leon
,
P. Stephen
Patrick
,
Adam
Doherty
,
Harry
Allan
,
Silvia
Cipiccia
,
Shashidhara
Marathe
,
Kaz
Wanelik
,
Michela
Esposito
,
Charlotte K.
Hagen
,
Alessandro
Olivo
,
Marco
Endrizzi
Diamond Proposal Number(s):
[30748]
Open Access
Abstract: Two-directional beam-tracking (2DBT) is a method for phase-contrast imaging and tomography that uses an intensity modulator to structure the X-ray beam into an array of independent circular beamlets that are resolved by a high-resolution detector. It features isotropic spatial resolution, provides two-dimensional phase sensitivity, and enables the three-dimensional reconstructions of the refractive index decrement, δ, and the attenuation coefficient, μ. In this work, the angular sensitivity and the spatial resolution of 2DBT images in a synchrotron-based implementation is reported. In its best configuration, angular sensitivities of ∼20 nrad and spatial resolution of at least 6.25 µm in phase-contrast images were obtained. Exemplar application to the three-dimensional imaging of soft tissue samples, including a mouse liver and a decellularized porcine dermis, is also demonstrated.
|
Sep 2024
|
|
I13-2-Diamond Manchester Imaging
|
Diamond Proposal Number(s):
[28574]
Open Access
Abstract: Beam tracking and edge illumination are phase contrast imaging techniques that rely on amplitude modulated x-ray beams to generate sensitivity to refraction and scattering. While each technique has its advantage (“single shot” three-contrast imaging in beam tracking; the ability to work with relatively large pixels in edge illumination), they also share a common drawback, namely that the modulator shields parts of the sample and, thus, prevents those areas from contributing to the image (under-sampling). Sample stepping, by which frames are acquired with the sample in a different position relative to the modulator (sometimes referred to as “dithering”) can produce well-sampled images. However, in computed tomography (CT), stepping must be performed at each rotation angle, enforcing step-and-shoot acquisitions and leading to long scan times. To enable faster acquisitions, fly scan compatible scanning schemes based on “roto-translating” the sample in the modulated x-ray beam were recently developed. This article reviews these schemes and provides practical guidance for their implementation.
|
May 2024
|
|
|
|
Adam
Doherty
,
Sylvain
Fourmaux
,
Alberto
Astolfo
,
Ralf F.
Ziesche
,
Jonathan
Wood
,
Oliver
Finlay
,
Wiebe
Stolpe
,
Darren
Batey
,
Ingo
Manke
,
François
Légaré
,
Matthieu
Boone
,
Dan
Symes
,
Zulfikar
Najmudin
,
Marco
Endrizzi
,
Alessandro
Olivo
,
Silvia
Cipiccia
Open Access
Abstract: Laser-plasma accelerators are compact linear accelerators based on the interaction of high-power lasers with plasma to form accelerating structures up to 1000 times smaller than standard radiofrequency cavities, and they come with an embedded X-ray source, namely betatron source, with unique properties: small source size and femtosecond pulse duration. A still unexplored possibility to exploit the betatron source comes from combining it with imaging methods able to encode multiple information like transmission and phase into a single-shot acquisition approach. In this work, we combine edge illumination-beam tracking (EI-BT) with a betatron X-ray source and present the demonstration of multimodal imaging (transmission, refraction, and scattering) with a compact light source down to the femtosecond timescale. The advantage of EI-BT is that it allows multimodal X-ray imaging technique, granting access to transmission, refraction and scattering signals from standard low-coherence laboratory X-ray sources in a single shot.
|
Oct 2023
|
|
I13-2-Diamond Manchester Imaging
|
Diamond Proposal Number(s):
[31421]
Open Access
Abstract: Background: Microscopic imaging of cartilage is a key tool for the study and development of treatments for osteoarthritis. When cellular and sub-cellular resolution is required, histology remains the gold standard approach, albeit limited by the lack of volumetric information as well as by processing artifacts. Cartilage imaging with the sub-cellular resolution has only been demonstrated in the synchrotron environment. Purpose: To provide a proof-of-concept demonstration of the capability of a laboratory-based x-ray phase-contrast microscope to resolve sub-cellular features in a cartilage sample. Methods: This work is based on a laboratory-based x-ray microscope using intensity-modulation masks. The structured nature of the beam, resulting from the mask apertures, allows the retrieval of three contrast channels, namely, transmission, refraction and dark-field, with resolution depending only on the mask aperture width. An ex vivo equine cartilage sample was imaged with the x-ray microscope and results were validated with synchrotron tomography and histology. Results: Individual chondrocytes, that is, cells responsible for cartilage formation, could be detected with the laboratory-based microscope. The complementarity of the three retrieved contrast channels allowed the detection of sub-cellular features in the chondrocytes. Conclusions: We provide the first proof-of-concept of imaging cartilage tissue with sub-cellular resolution using a laboratory-based x-ray microscope.
|
Jul 2023
|
|
I13-1-Coherence
|
Savvas N.
Savvides
,
Mattia F.
Gerli
,
Antonio
Citro
,
Lorenzo
Massimi
,
Charlotte K.
Hagen
,
Marco
Endrizzi
,
Alessia
Atzeni
,
Alberto
Astolfo
,
Michela
Esposito
,
Olumide K.
Ogunbiyi
,
Mark
Turmaine
,
Elizabeth S.
Smith
,
Silvia
Cipiccia
,
Christoph
Rau
,
Peng
Li
,
Roberto
Lutman
,
Giulia
Selmin
,
Natalie
Durkin
,
Soichi
Shibuya
,
Marianna
Scuglia
,
Marco
Pellegrini
,
Paolo
De Coppi
,
Alessandro
Olivo
Abstract: Tissue engineering (TE) holds promise for generating lab-grown patient specific organs which can provide: (1) effective treatment for conditions that require volumetric tissue transplantation and (2) new platforms for drug testing. Even though volumetric structural information is essential for confirming successful organ maturation, TE protocol designs are currently informed through destructive and 2D construct assessment tools (e.g. histology). X-ray phase-contrast computed-tomography (PC-CT) can generate non-destructive, high resolution, 3D density maps of organ architecture. In this work, PC-CT is used as new imaging tool for guiding two TE protocols currently at the in-vitro testing stage. The first (1) involves cell-repopulation of an oesophageal scaffold, with the aim of using the regenerated construct for treating long-gap oesophageal atresia, whilst for the second (2) a lung-derived scaffold is populated with islets for regenerating a pancreas, with the “repurposed” lung offering a platform for diabetes drug testing. By combing 3D images and quantitative information, we were able to perform comprehensive construct evaluation. Specifically, we assessed volumetrically: (1) the cell-distribution within the regenerated oesophagi and (2) islet integration with the vascular tree of the lung-derived scaffold. This new information was proven to be essential for establishing corresponding TE protocols and enabled their progression to more advanced scale-up models. We are confident that PC-CT will provide the novel insights necessary to further progress TE protocols, with the next step being in-vivo testing. Crucially, the non-destructive nature of PC-CT will allow in-vivo assessments of TE constructs following their implantation into animal hosts, to investigate their successful integration.
|
Apr 2023
|
|
