scholarly journals Three-Dimensional Super-Resolved Imaging of Paraffin-Embedded Kidney Samples

Kidney360 ◽  
2021 ◽  
pp. 10.34067/KID.0005882021
Author(s):  
David Unnersjö-Jess ◽  
Amer Ramdedovic ◽  
Martin Höhne ◽  
Linus Butt ◽  
Felix C. Koehler ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Kidney Tissue ◽  
Confocal Imaging ◽  
Disease Assessment ◽  
Preparation Methods ◽  
Gold Standard Method ◽  
Filtration Barrier ◽  
Disease Entities ◽  
Kidney Imaging ◽  
Monitoring Treatment

Background: Diseases of the glomeruli, the renal filtration units, are a leading cause of progressive kidney disease. Assessment of the ultrastructure of podocytes at the glomerular filtration barrier is essential for diagnosing diverse disease entities, providing insight into the disease pathogenesis as well as monitoring treatment responses. Methods: We here apply previously published sample preparation methods together with STED and confocal microscopy for resolving nanoscale podocyte substructure. The protocols are modified and optimized in order to be applied to samples which have been formalin fixed and paraffin-embedded (FFPE). Results: We successfully modify our protocols to allow for deep three-dimensional STED and confocal imaging of FFPE kidney tissue with similar staining and image quality as compared to our previous approaches. We further show that quantitative analysis can be applied to extract morphometrics of healthy and diseased samples from both mice and humans. Conclusions: The results from this study could increase the feasibility to implement optical kidney imaging protocols in clinical routines, as FFPE is the gold standard method for storage of patient samples.

scholarly journals Three-dimensional Super-resolved Imaging of Paraffin-embedded Kidney Samples

2021 ◽  
Author(s):  
David Unnersjoe-Jess ◽  
Amer Ramdedovic ◽  
Martin Hoehne ◽  
Linus Butt ◽  
Felix C Koehler ◽  
...  
Keyword(s):  
New Technologies ◽  
Three Dimensional ◽  
Kidney Tissue ◽  
Confocal Imaging ◽  
Disease Assessment ◽  
Gold Standard Method ◽  
Filtration Barrier ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Monitoring Treatment

Diseases of the glomeruli, the renal filtration units, are a leading cause of progressive kidney disease. Assessment of the ultrastructure of podocytes at the glomerular filtration barrier is essential for diagnosing diverse disease entities, providing insight into the disease pathogenesis as well as monitoring treatment responses. New technologies, including super-resolved nanoscopy and expansion microscopy, as well as new sample preparation techniques, are starting to revolutionize imaging of biopsy specimens. However, our previous approaches for simple and fast three-dimensional imaging of optically cleared samples are to date not compatible with formalin fixed paraffin-embedded (FFPE) tissue, impeding application in clinical routine. Here we provide protocols that circumvent these limitations and allow for three dimensional STED and confocal imaging of FFPE kidney tissue with similar staining and image quality as compared to our previous approaches. This would increase the feasibility to implement these protocols in clinical routines, as FFPE is the gold standard method for storage of patient samples.

scholarly journals First Asian population study of stereotactic body radiation therapy for ventricular arrhythmias

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Li-Ting Ho ◽  
Jenny Ling-Yu Chen ◽  
Hsing-Min Chan ◽  
Yu-Cheng Huang ◽  
Mao-Yuan Su ◽  
...  
Keyword(s):  
Radiation Therapy ◽  
Three Dimensional ◽  
Asian Population ◽  
Target Volume ◽  
Dual Energy ◽  
Detection Sensitivity ◽  
Dual Energy Ct ◽  
Stereotactic Body Radiation ◽  
Integrated Boost ◽  
Monitoring Treatment

AbstractWe report the first Asian series on stereotactic body radiation (SBRT) for refractory ventricular arrhythmia (VA) in Taiwanese patients. Three-dimensional electroanatomic maps, delayed-enhancement magnetic resonance imaging (DE-MRI), and dual-energy computed tomography (CT) were used to identify scar substrates. The main target volume was treated with a single radiation dose of 25 Gy and the margin volume received 20 Gy using simultaneous integrated boost delivered by the Varian TrueBeam system. Efficacy was assessed according to VA events recorded by an implantable cardioverter-defibrillator (ICD) or a 24-h Holter recorder. Pre- and post-radiation therapy imaging studies were performed. From February 2019 to December 2019, seven patients (six men, one woman; mean age, 55 years) were enrolled and treated. One patient died of hepatic failure. In the remaining six patients, at a median follow-up of 14.5 months, the VA burden and ICD shocks significantly decreased (only one patient with one ICD shock after treatment). Increased intensity on DE-MRI might be associated with a lower risk for VA recurrence, whereas dual-energy CT had lower detection sensitivity. No acute or minimal late adverse events occurred. In patients with refractory VA, SBRT is associated with a marked reduction in VA burden and ICD shocks, and DE-MRI might be useful for monitoring treatment effects.

Hydrogel-Based Bioinks for Three-Dimensional Bioprinting: Patent Analysis

2021 ◽  
Vol 7 (1) ◽  
pp. 3
Author(s):  
Ahmed Fatimi
Keyword(s):  
Tissue Engineering ◽  
Detailed Analysis ◽  
State Of The Art ◽  
Three Dimensional ◽  
Patent Analysis ◽  
The State ◽  
Driving Factors ◽  
3D Bioprinting ◽  
Preparation Methods

There are a variety of hydrogel-based bioinks commonly used in three-dimensional bioprinting. In this study, in the form of patent analysis, the state of the art has been reviewed by introducing what has been patented in relation to hydrogel-based bioinks. Furthermore, a detailed analysis of the patentability of the used hydrogels, their preparation methods and their formulations, as well as the 3D bioprinting process using hydrogels, have been provided by determining publication years, jurisdictions, inventors, applicants, owners, and classifications. The classification of patents reveals that most inventions intended for hydrogels used as materials for prostheses or for coating prostheses are characterized by their function or properties Knowledge clusters and expert driving factors show that biomaterials, tissue engineering, and biofabrication research is concentrated in the most patents.

scholarly journals Bowel preparation for performing a colonoscopy: prospective randomized comparison study between a low-volume solution of polyethylene glycol and bisacodyl versus bisacodyl and a mannitol solution

2012 ◽  
Vol 49 (2) ◽  
pp. 162-168 ◽  
Author(s):  
Manoel Carlos Vieira ◽  
Claudio Lyoiti Hashimoto ◽  
Flair José Carrilho
Keyword(s):  
Polyethylene Glycol ◽  
Vital Signs ◽  
Abdominal Distension ◽  
Similar Efficacy ◽  
Preparation Methods ◽  
Gold Standard Method ◽  
Mannitol Solution ◽  
Before And After ◽  
Glycol Solution

CONTEXT: Colonoscopy is currently the gold standard method to examine the colon, the rectum and the terminal ileum. In order to perform the colonoscopy, it is necessary to clean the bowel and use medications that are generally poorly tolerated by the patients. OBJECTIVE: Compare the tolerability, acceptability, safety and efficacy of two solutions used for intestinal preparation for a colonoscopy. METHODS: One hundred patients matched for sex and age were prospective randomized into two groups. Polyethylene glycol group received bisacodyl 10 mg plus 1 L of polyethylene glycol the night before and 1 L on the day of the exam. Mannitol group received bisacodyl 20 mg the day before and 1 L of a 10% mannitol solution on the day of the exam. The diet was the same for both groups. Tolerability and acceptability were measured using previously validated questionnaires. In terms of safety, variations in vital signs before and after the preparation were recorded, in addition to any complications. The quality of the preparation was graded based on the Boston and Ottawa scales. RESULTS: Ninety-six percent (96%) completed the study. As for tolerability, the mannitol preparation group exhibited a significantly higher frequency of nausea, vomiting, abdominal pain, and abdominal distension than polyethylene glycol group (P < 0.05). Acceptability was significantly better in polyethylene glycol group. The polyethylene glycol solution has also previously been shown to be safer than mannitol. No difference was observed in the quality of the preparation between the two preparation methods. CONCLUSIONS: The following conclusions can be made: polyethylene glycol solution had higher tolerability, acceptability, and safety than the mannitol and should be used instead of mannitol. Both preparation solutions have similar efficacy.

scholarly journals Three-Dimensional High-Resolution Digital Inline Hologram Reconstruction with a Volumetric Deconvolution Method

Sensors ◽  
2018 ◽  
Vol 18 (9) ◽  
pp. 2918 ◽  
Author(s):  
Junseong Eom ◽  
Sangjun Moon
Keyword(s):  
High Resolution ◽  
Whole Blood ◽  
Tissue Sample ◽  
Three Dimensional ◽  
Kidney Tissue ◽  
Wide Field ◽  
Deconvolution Technique ◽  
Micron Size ◽  
Volumetric Images ◽  
Focus Image

The digital in-line holographic microscope (DIHM) was developed for a 2D imaging technology and has recently been adapted to 3D imaging methods, providing new approaches to obtaining volumetric images with both a high resolution and wide field-of-view (FOV), which allows the physical limitations to be overcome. However, during the sectioning process of 3D image generation, the out-of-focus image of the object becomes a significant impediment to obtaining evident 3D features in the 2D sectioning plane of a thick biological sample. Based on phase retrieved high-resolution holographic imaging and a 3D deconvolution technique, we demonstrate that a high-resolution 3D volumetric image, which significantly reduces wave-front reconstruction and out-of-focus artifacts, can be achieved. The results show a 3D volumetric image that is more finely focused compared to a conventional 3D stacked image from 2D reconstructed images in relation to micron-size polystyrene beads, a whole blood smear, and a kidney tissue sample. We believe that this technology can be applicable for medical-grade images of smeared whole blood or an optically cleared tissue sample for mobile phytological microscopy and laser sectioning microscopy.

Three-dimensional graphic reconstruction of the insect exoskeleton through confocal imaging of endogenous fluorescence

2000 ◽  
Vol 48 (6) ◽  
pp. 367-384 ◽  
Author(s):  
Sasha Zill ◽  
S. Faith Frazier ◽  
David Neff ◽  
Laura Quimby ◽  
Michelle Carney ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Confocal Imaging ◽  
Endogenous Fluorescence

scholarly journals The Influence of Astaxanthin on the Proliferation of Adipose-derived Mesenchymal Stem Cells in Gelatin-Methacryloyl (GelMA) Hydrogels

Materials ◽  
2019 ◽  
Vol 12 (15) ◽  
pp. 2416 ◽  
Author(s):  
Bo Young Choi ◽  
Elna Paul Chalisserry ◽  
Myoung Hwan Kim ◽  
Hyun Wook Kang ◽  
Il-Whan Choi ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cellular Proliferation ◽  
Three Dimensional ◽  
Confocal Imaging ◽  
Alamar Blue Assay ◽  
Ratio Test ◽  
Alamar Blue ◽  
Chemical Structures

Recently, astaxanthin, a red lipophilic pigment belonging to the xanthophyllic family of carotenoids, has shown the feasibility of its uses in tissue engineering and regenerative medicine, due to its excellent antioxidant activities and its abilities to enhance the self-renewal potency of stem cells. In this study, we demonstrate the influence of astaxanthin on the proliferation of adipose-derived mesenchymal stem cells in tissue-engineered constructs. The tissue engineered scaffolds were fabricated using photopolymerizable gelatin methacryloyl (GelMA) with different concentrations of astaxanthin. The effects of astaxanthin on cellular proliferation in two-dimensional environments were assessed using alamar blue assay and reverse transcription polymerase chain reaction (RT-PCR). Then, rheological properties, chemical structures and the water absorption of the fabricated astaxanthin-incorporated GelMA hydrogels were characterized using NMR analysis, rheological analysis and a swelling ratio test. Finally, the influence in three-dimensional environments of astaxanthin-incorporated GelMA hydrogels on the proliferative potentials of adipose-derived stem cells was assessed using alamar blue assay and the confocal imaging with Live/dead staining. The experimental results of the study indicate that an addition of astaxanthin promises to induce stem cell potency via proliferation, and that it can be a useful tool for a three-dimensional culture system and various tissue engineering applications.

scholarly journals Renal reabsorption in 3D vascularized proximal tubule models

2019 ◽  
Vol 116 (12) ◽  
pp. 5399-5404 ◽  
Author(s):  
Neil Y. C. Lin ◽  
Kimberly A. Homan ◽  
Sanlin S. Robinson ◽  
David B. Kolesky ◽  
Nathan Duarte ◽  
...  
Keyword(s):  
Proximal Tubule ◽  
Disease Modeling ◽  
Three Dimensional ◽  
Kidney Tissue ◽  
Renal Reabsorption ◽  
Native Kidney ◽  
Glucose Reabsorption ◽  
Diseased State ◽  
And Function

Three-dimensional renal tissues that emulate the cellular composition, geometry, and function of native kidney tissue would enable fundamental studies of filtration and reabsorption. Here, we have created 3D vascularized proximal tubule models composed of adjacent conduits that are lined with confluent epithelium and endothelium, embedded in a permeable ECM, and independently addressed using a closed-loop perfusion system to investigate renal reabsorption. Our 3D kidney tissue allows for coculture of proximal tubule epithelium and vascular endothelium that exhibits active reabsorption via tubular–vascular exchange of solutes akin to native kidney tissue. Using this model, both albumin uptake and glucose reabsorption are quantified as a function of time. Epithelium–endothelium cross-talk is further studied by exposing proximal tubule cells to hyperglycemic conditions and monitoring endothelial cell dysfunction. This diseased state can be rescued by administering a glucose transport inhibitor. Our 3D kidney tissue provides a platform for in vitro studies of kidney function, disease modeling, and pharmacology.

scholarly journals Contributions of adipose tissue architectural and tensile properties toward defining healthy and unhealthy obesity

2014 ◽  
Vol 306 (3) ◽  
pp. E233-E246 ◽  
Author(s):  
Denise E. Lackey ◽  
David H. Burk ◽  
Mohamed R. Ali ◽  
Rouzbeh Mostaedi ◽  
William H. Smith ◽  
...  
Keyword(s):  
Tensile Strength ◽  
Adipose Tissue ◽  
Proteolytic Enzymes ◽  
Three Dimensional ◽  
Confocal Imaging ◽  
Obese Women ◽  
Multivariate Statistical ◽  
Related Factors ◽  
Ecm Remodeling ◽  
Stress Signals

The extracellular matrix (ECM) plays an important role in the maintenance of white adipose tissue (WAT) architecture and function, and proper ECM remodeling is critical to support WAT malleability to accomodate changes in energy storage needs. Obesity and adipocyte hypertrophy place a strain on the ECM remodeling machinery, which may promote disordered ECM and altered tissue integrity and could promote proinflammatory and cell stress signals. To explore these questions, new methods were developed to quantify omental and subcutaneous WAT tensile strength and WAT collagen content by three-dimensional confocal imaging, using collagen VI knockout mice as a methods validation tool. These methods, combined with comprehensive measurement of WAT ECM proteolytic enzymes, transcript, and blood analyte analyses, were used to identify unique pathophenotypes of metabolic syndrome and type 2 diabetes mellitus in obese women, using multivariate statistical modeling and univariate comparisons with weight-matched healthy obese individuals. In addition to the expected differences in inflammation and glycemic control, approximately 20 ECM-related factors, including omental tensile strength, collagen, and enzyme transcripts, helped discriminate metabolically compromised obesity. This is consistent with the hypothesis that WAT ECM physiology is intimately linked to metabolic health in obese humans, and the studies provide new tools to explore this relationship.

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