Tag: synchrotron imaging

New strategy combines techniques to study intracellular transport of nanoparticles

New strategy combines techniques to study intracellular transport of nanoparticles

Research led by CNPEM scientists reveals intracellular movement of nanoparticles coated with a protein corona. The technique employed in the study will be available at the Sibipiruna beamline, which is dedicated to research on BSL-4 pathogens.

Researchers from the Brazilian Center for Research in Energy and Materials (CNPEM), in collaboration with institutions in Brazil, the United Kingdom and the United States, have demonstrated a new strategy to monitor the intracellular trajectory of nanoparticles. The study was one of the cover features for the journal Small in June 2025, and combined different high-resolution microscopy techniques to observe how these particles move around in the cellular environment over time.

The research used advanced microscopy resources from the Nanotechnology National Laboratory (LNNano) and Sirius facilities, and included a technique not yet available at the center, X-ray cryotomography. Measurements were obtained using a beamline with characteristics similar to the future Sibipiruna line, which will be part of Project Orion. The resulting data verified the use of this technique in cryogenic conditions, as well as its ability to reveal cellular structures smaller than the viruses that will be studied in the future laboratory complex. 

The approach made it possible to identify the migration of nanoparticles to the perinuclear region of cells and fusion of the vesicles that transport them, without the use of contrast agents. The results overcome common limitations in studies of this type, and offer a promising tool for understanding how nanomaterials behave in complex biological systems.  

Nanoparticles and the challenge of cell internalization

Nanoparticles have been widely studied for their potential in biomedical applications such as controlled release of medications, diagnostic imaging and targeted therapy. But these applications still face major obstacles, especially with regard to detailed understanding of the mechanisms through which these particles are internalized and move within cells. 

The formation of the protein corona, a layer of biomolecules that adsorbs onto the surface of nanoparticles when they come into contact with biological fluids, is a good example of the complexity involved in investigating the mechanisms of internalization and intracellular transit. This layer significantly alters the physical and chemical properties of the particles, influencing their stability and interaction with different cell types. Understanding the behavior of these nanoparticles in the intracellular environment consequently requires approaches that take into account both cell dynamics and the variability introduced by the corona’s composition. 

Despite advances in characterization techniques, most studies offer only specific or static views of the internalization process; it is generally not possible to distinguish between particles absorbed at different times, or to follow their precise location within the cell over time. This study conducted by CNPEM researchers proposes an alternative approach intended to overcome these obstacles through an experimental strategy that employs different imaging methods, providing a broader analysis that extracts the best from each technique. 

A new approach to studying cell dynamics

The researchers proposed a protocol based on a short period of cell exposure, followed by complete removal of the unabsorbed nanoparticles and cryopreservation of the cells after different time intervals (0, 2, and 24 hours). Nanoparticle internalization by the cells was then evaluated using wide-field fluorescence microscopy, and showed progressive migration to the perinuclear region. 

“Previous studies investigating the internalization process of nanoparticles also used cells that were fixed after different time intervals but incubated continuously. As a result of this method, nanoparticles internalized at the beginning of the incubation period cannot be distinguished from those internalized at the end. The alternative method we are proposing avoids this problem and facilitates analysis of the sequence of events and changes associated with the cell internalization process,” explains Mateus Cardoso, an author of the article and researcher at the Synchrotron Light National Laboratory (LNLS), which is part of CNPEM. 

Read more on CNPEM website

Image: FFibroblasts after incubation with silica nanoparticles in the presence of bovine serum albumin (BSA). Wide-field fluorescence microscopy image. (Image adapted from Galdino et al., Small, 2024, https://doi.org/10.1002/smll.202409065)

Biomaterials for Improving Ovarian Tissue Transplantation

Biomaterials for Improving Ovarian Tissue Transplantation

Cryopreservation of ovarian tissue and its subsequent transplantation represent a big hope to preserve fertility in young women who have defeated cancer. Ovary revascularization is a crucial factor impacting the outcome of the engraftment. Limited oxygenation may have severe consequences on the ovarian reserve, with a significant loss of follicles. New frontiers in reproductive technology aim to reduce the ischemic/hypoxic window following auto-transplantation procedures. Biomaterials supplemented with ovarian-derived endothelial cells could be the solution to enhance vascular regeneration in the transplanted tissue.

In this study, we propose a combined Advanced Therapeutic Medicinal Product (ATMP) obtained from the association of cryopreserved ovarian tissue with a 3D dermal substitute — a biocompatible and bioactive scaffold employed in regenerative medicine — pre-seeded with vascular system cells previously isolated from the same ovarian tissue. This pre-seeding, known as inosculation, is a bioengineering approach aimed at enhancing revascularization by promoting the formation of novel vascular networks within the scaffold prior to implantation. The goal of the research is to demonstrate that a such graft can boost the growth of new vessels (Fig. 1), potentially improving the ovary survival and functionality.

To evaluate the effectiveness of this approach, several techniques were employed including synchrotron radiation-based X-ray phase-contrast microtomography (SR PC-microCT). As a volumetric imaging technique, SR PC-microCT enables three-dimensional visualization of the inner anatomical structures of the proposed ATMP at high spatial and contrast resolution, with the additional advantage of being non-destructive. Scans were carried out at the SYRMEP Imaging beamline of Elettra. The findings obtained by the X-ray images were complemented by histology and immunohistochemical analyses, adhesion and proliferation assays, gene expression and immunofluorescence. 

A bovine collagen-based scaffold, Integra®, was selected among various dermal substitute materials tested and was used as a support for ovarian transplantation in subsequent in vivo experiments on mouse models. Histology clearly demonstrates the presence of endothelial cells within the Integra® matrix, exhibiting a tendency to form vascular structures. Red-blood cells can be also observed inside the developing vessels (Fig. 2a). Similarly, Fig. 2b shows a virtual slice obtained by X-ray PC microCT of a sample region at the interface between the ovarian tissue and the Integra® support. In agreement with the histological data, the X-ray image shows a massive accumulation of dense structures within the scaffold, which may be attributed to a high concentration of endothelial cells. Notably, SR PC microCT enables the cells distribution within the scanned blocks to be tracked, revealing a migration of the endothelial cells from the matrix into the tissue with a preferential side of accumulation. Supplementary videos are available on the full paper website (please follow the link at bottom of this page).

Read more on Elettra website

 

Scotty’s rib: University of Regina PhD student examines preserved blood vessels in famous fossil

Scotty’s rib: University of Regina PhD student examines preserved blood vessels in famous fossil

A University of Regina research team made some dino-mite discoveries about how dinosaurs may have healed from injuries when they examined the preserved blood vessel structures inside a rib bone from Scotty, the famous Tyrannosaurus rex unearthed in Saskatchewan in the 1990s. Their findings were recently published in Scientific Reports, an open-access journal that publishes original research from across the natural sciences, psychology, medicine, and engineering.

Jerit L. Mitchell, a PhD student in the Department of Physics at the University of Regina, is the study’s lead author, who joined the research project in 2019 when Scotty’s rib was scanned at the Canadian Light Source (CLS) for the first time. Mitchell was finishing his undergraduate honours thesis research when he discovered the vessel structures.

“I remember showing my supervisors, Dr. Barbi and Dr. McKellar, a strange structure inside a scan of the rib that I originally didn’t give much thought to. They were quick to point out that what I discovered could possibly be preserved blood vessels, which has since led to a much more expansive research project,” says Mitchell.

The powerful synchrotron X-rays produced by the CLS at the University of Saskatchewan enabled the researchers to create a detailed 3D model of both the T. rex bone and the soft tissue structures that reside inside without damaging the 66-million-year-old fossil. Then, using chemical analysis, the researchers determined what elements and molecules make up the vessel structures, allowing them to hypothesize how the structures were preserved over millions of years.

The X-rays of Scotty’s rib showed a healed fracture, possibly due to a fight with another dinosaur. This discovery could provide important, evolutionary information to researchers, such as the healing potential of a T. rex.

“It was a real treat to be able to contribute to this research,” says Mohsen Shakouri, a staff scientist at the CLS. “We are pleased that our ultrabright synchrotron light helped the team gain new insights into the physiology of everyone’s favorite T. rex.”

Dr. Mauricio Barbi, physics professor at the U of R and Mitchell’s graduate supervisor, says this discovery could help focus the search for soft tissue in fossils.

“Preserved blood vessel structures, like we have found in Scotty’s rib bone, appear linked to areas where the bone was healing. This is because during the healing process, those areas had increased blood flow to them,” says Barbi. “This work also provides a new way to compare how injuries healed in extinct animals, like dinosaurs, with living species, such as birds and reptiles, which helps us better understand the biology of the past, and also how life on Earth has evolved over millions of years.”

Read more on CLS website

Last meal reveals eating habits of Australian sauropod

Last meal reveals eating habits of Australian sauropod

Key Points

  • Research led by Curtin University has confirmed that at least some sauropods were plant eaters
  • The investigation is believed to be the first identification of gut contents in a sauropod
  • Advanced imaging techniques at the Australian Synchrotron and Australian Centre for Neutron Scattering provided supporting evidence of the gut content

International research led by Curtin University and supported by ANSTO, has identified and studied the first sauropod dinosaur gut contents found anywhere in the world. The stomach content was preserved with a reasonably complete skeleton of the Australian Cretaceous species Diamantinasaurus matildae found in Winton Queensland.

Imaging on the Imaging and Medical beamline at the Australian Synchrotron and the neutron tomography instrument Dingo at the Australian Centre for Neutron Scattering provided supporting evidence of the stomach contents, known as a cololite, and the first sauropod skin found in Australia, which was found associated with the cololite. 

The investigative team, led by Curtin University and Australian Age of Dinosaurs Museum of Natural History, included ANSTO, University College London (UK), University of Colorado Boulder (US), University of New England (Aus), Swedish Museum of Natural History, and CSIRO. 

“Our interpretation of this specimen as the preservation of a genuine cololite rests on the physical properties of the rock, studies of the burial decay and preservation processes that affect animal and plant remains as they become fossilised, micro-computed tomographic, and geochemical evidence, as well as the abundance of plants within the cololite relative to other rocks in the site,” explained Dr Jospeh Bevitt, a co-author on the paper published in Current Biology.

“Sauropod dinosaurs are iconic, unmistakable, and include among their ranks the largest terrestrial animals of all time, including Brontosaurus, Brachiosaurus, and Argentinasaurus ,” he added.

“There has been little doubt since the 1870s that they were herbivorous, or plant-eating. However, the specific plants eaten by sauropods, and the heights above ground at which they fed, have remained obscure. This is because of the lack of direct fossil evidence in the form of gut contents— until now,” said Dr Bevitt, an expert in analysing fossilised remains of dinosaurs and other prehistoric fossils.

The Diamantinasaurus cololite allowed the research team to draw several overarching conclusions. At least some sauropods were herbivorous, a confirmation that supported more than 150 years of scientific theory. 

Sauropods did not engage in much processing of food in their mouths, which also supported scientific consensus. 

Read more on ANSTO website

Improving steel pipelines for safe transport of hydrogen

Improving steel pipelines for safe transport of hydrogen

USask researchers use synchrotron light to capture 3D images of cracks that form inside steel.

Hydrogen is increasingly gaining attention as a promising energy source for a cleaner, more sustainable future. Using hydrogen to meet the energy demands for large-scale applications such as utility infrastructure will require transporting large volumes via existing pipelines designed for natural gas.

But there’s a catch. Hydrogen can weaken the steel that these pipelines are made of. When hydrogen atoms enter the steel, they diffuse into its microstructure and can cause the metal to become brittle, making it more susceptible to cracking. Hydrogen can be introduced into the steel during manufacturing, or while the pipeline is in service transporting oil and gas.

To better understand this problem, researcher Tonye Jack used the Canadian Light Source (CLS) at the University of Saskatchewan (USask) to capture a 3D view of the cracks formed in steels. Researchers have previously relied on two-dimensional imaging techniques, which don’t provide the same rich detail made possible with synchrotron radiation.

Tonye, a PhD candidate in USask’s Department of Mechanical Engineering, and his colleagues studied different pipeline steels and showed that microstructure plays a critical role in how much hydrogen the steel absorbs and how it is distributed in the metal. Their research also revealed that when hydrogen enters the steel while the pipeline is in service, it causes more damage than if introduced during manufacturing or other pre-charging conditions.

The risk of steel failure due to hydrogen embrittlement depends on several factors such as the amount of hydrogen in the steel, the steel’s microstructure, stress conditions, and operating environment. However, Tonye emphasizes that how much hydrogen is retained in the steel and where it accumulates largely dictates its failure behavior.

“We need to know the mechanism of failure and how to mitigate it,” he says.

While catastrophic pipeline failures are rare, his team’s findings are important as industries plan to transport hydrogen gas using high-strength natural gas pipelines. “These findings can help inform the production of safer pipelines,” he says. By refining the microstructure, manufacturers can design steels that are more resistant to cracking and hydrogen embrittlement.

Read more on CLS website