scholarly journals Computational 4D-OCM for label-free imaging of collective cell invasion and force-mediated deformations in collagen

2020 ◽  
Author(s):  
Jeffrey A. Mulligan ◽  
Lu Ling ◽  
Claudia Fischbach ◽  
Steven G. Adie
Keyword(s):  
Cancer Progression ◽  
Imaging Techniques ◽  
Traction Force ◽  
Time Lapse ◽  
Label Free ◽  
Imaging Data ◽  
Scattering Media ◽  
Current Standard ◽  
Isolated Cells ◽  
Large Tumor

AbstractTraction force microscopy (TFM) is an important family of techniques used to measure and study the role of cellular traction forces (CTFs) associated with many biological processes. However, current standard TFM methods rely on imaging techniques that do not provide the experimental capabilities necessary to study CTFs within 3D collective and dynamic systems embedded within optically scattering media. Traction force optical coherence microscopy (TF-OCM) was developed to address these needs, but has only been demonstrated for the study of isolated cells embedded within optically clear media. Here, we present computational 4D-OCM methods that enable the study of dynamic invasion behavior of large tumor spheroids embedded in collagen. Our multi-day, time-lapse imaging data provided detailed visualizations of evolving spheroid morphology, collagen degradation, and collagen deformation, all using label-free scattering contrast. These capabilities, which provided insights into how stromal cells affect cancer progression, significantly expand access to critical data about biophysical interactions of cells with their environment, and lay the foundation for future efforts toward volumetric, time-lapse reconstructions of collective CTFs with TF-OCM.

scholarly journals Computational 4D-OCM for label-free imaging of collective cell invasion and force-mediated deformations in collagen

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jeffrey A. Mulligan ◽  
Lu Ling ◽  
Nichaluk Leartprapun ◽  
Claudia Fischbach ◽  
Steven G. Adie
Keyword(s):  
Cancer Progression ◽  
Imaging Techniques ◽  
Traction Force ◽  
Time Lapse ◽  
Label Free ◽  
Imaging Data ◽  
Scattering Media ◽  
Current Standard ◽  
Isolated Cells ◽  
Large Tumor

AbstractTraction force microscopy (TFM) is an important family of techniques used to measure and study the role of cellular traction forces (CTFs) associated with many biological processes. However, current standard TFM methods rely on imaging techniques that do not provide the experimental capabilities necessary to study CTFs within 3D collective and dynamic systems embedded within optically scattering media. Traction force optical coherence microscopy (TF-OCM) was developed to address these needs, but has only been demonstrated for the study of isolated cells embedded within optically clear media. Here, we present computational 4D-OCM methods that enable the study of dynamic invasion behavior of large tumor spheroids embedded in collagen. Our multi-day, time-lapse imaging data provided detailed visualizations of evolving spheroid morphology, collagen degradation, and collagen deformation, all using label-free scattering contrast. These capabilities, which provided insights into how stromal cells affect cancer progression, significantly expand access to critical data about biophysical interactions of cells with their environment, and lay the foundation for future efforts toward volumetric, time-lapse reconstructions of collective CTFs with TF-OCM.

scholarly journals Three-dimensional label-free observation of individual bacteria upon antibiotic treatment using optical diffraction tomography

2019 ◽  
Author(s):  
Jeonghun Oh ◽  
Jea Sung Ryu ◽  
Moosung Lee ◽  
Jaehwang Jung ◽  
Seung yun Han ◽  
...  
Keyword(s):  
Imaging Techniques ◽  
Three Dimensional ◽  
Time Lapse ◽  
Optical Diffraction ◽  
Response Analysis ◽  
Diffraction Tomography ◽  
Label Free ◽  
Optical Diffraction Tomography ◽  
Quantitative Manner ◽  
Basic Studies

AbstractMeasuring alterations in bacteria upon antibiotic application is important for basic studies in microbiology, drug discovery, and clinical diagnosis, and disease treatment. However, imaging and 3D time-lapse response analysis of individual bacteria upon antibiotic application remain largely unexplored mainly due to limitations in imaging techniques. Here, we present a method to systematically investigate the alterations in individual bacteria in 3D and quantitatively analyze the effects of antibiotics. Using optical diffraction tomography, in-situ responses of Escherichia coli and Bacillus subtilis to various concentrations of ampicillin were investigated in a label-free and quantitative manner. The presented method reconstructs the dynamic changes in the 3D refractive-index distributions of living bacteria in response to antibiotics at sub-micrometer spatial resolution.

scholarly journals OrBITS: A High-throughput, time-lapse, and label-free drug screening platform for patient-derived 3D organoids

2021 ◽  
Author(s):  
Christophe Deben ◽  
Edgar Cardenas De La Hoz ◽  
Maxim Le Compte ◽  
Paul Van Schil ◽  
Jeroen M. Hendriks ◽  
...  
Keyword(s):  
Personalized Medicine ◽  
High Throughput ◽  
Basic Research ◽  
Automated Analysis ◽  
Time Lapse ◽  
Industrial Applications ◽  
Great Promise ◽  
Label Free ◽  
Current Standard ◽  
Convolutional Network

AbstractPatient-derived organoids are invaluable for fundamental and translational cancer research and holds great promise for personalized medicine. However, the shortage of available analysis methods, which are often single-time point, severely impede the potential and routine use of organoids for basic research, clinical practise, and pharmaceutical and industrial applications. Here, we report the development of a high-throughput automated organoid analysis platform that allows for kinetic monitoring of organoids, named Organoid Brightfield Identification-based Therapy Screening (OrBITS). The combination of computer vision with a convolutional network machine learning approach allowed for the detection and tracking of organoids in routine extracellular matrix domes, advanced Gri3D®-96 well plates, and high-throughput 384-well microplates, solely based on brightfield imaging. We used OrBITS to screen chemotherapeutics and targeted therapies, and incorporation of a fluorescent cell death marker, revealed further insight into the mechanistic action of the drug, a feature not achievable with the current gold standard ATP-assay. This manuscript describes the validation of the OrBITS deep learning analysis approach against current standard assays for kinetic imaging and automated analysis of organoids. OrBITS, as a scalable, high-throughput technology, would facilitate the use of patient-derived organoids for drug development, therapy screening, and guided clinical decisions for personalized medicine. The developed platform also provides a launching point for further brightfield-based assay development to be used for fundamental research.

Cleavage of Human Embryos: Options and Diversity

Acta Naturae ◽  
2016 ◽  
Vol 8 (3) ◽  
pp. 88-96
Author(s):  
Yu. K. Doronin ◽  
I. V. Senechkin ◽  
L. V. Hilkevich ◽  
M. A. Kurcer
Keyword(s):  
Time Lapse ◽  
Reproductive Technologies ◽  
Human Embryos ◽  
Imaging Data ◽  
Noninvasive Methods ◽  
Topographic Features ◽  
Time Parameters ◽  
Embryo Cleavage ◽  
Time Lapse Imaging ◽  
Developmental Dynamics

In order to estimate the diversity of embryo cleavage relatives to embryo progress (blastocyst formation), time-lapse imaging data of preimplantation human embryo development were used. This retrospective study is focused on the topographic features and time parameters of the cleavages, with particular emphasis on the lengths of cleavage cycles and the genealogy of blastomeres in 2- to 8-cell human embryos. We have found that all 4-cell human embryos have four developmental variants that are based on the sequence of appearance and orientation of cleavage planes during embryo cleavage from 2 to 4 blastomeres. Each variant of cleavage shows a strong correlation with further developmental dynamics of the embryos (different cleavage cycle characteristics as well as lengths of blastomere cycles). An analysis of the sequence of human blastomere divisions allowed us to postulate that the effects of zygotic determinants are eliminated as a result of cleavage, and that, thereafter, blastomeres acquire the ability of own syntheses, regulation, polarization, formation of functional contacts, and, finally, of specific differentiation. This data on the early development of human embryos obtained using noninvasive methods complements and extend our understanding of the embryogenesis of eutherian mammals and may be applied in the practice of reproductive technologies.

scholarly journals Time lapse synchrotron IR chemical imaging for observing the acclimation of a single algal cell to CO2 treatment

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ghazal Azarfar ◽  
Ebrahim Aboualizadeh ◽  
Simona Ratti ◽  
Camilla Olivieri ◽  
Alessandra Norici ◽  
...  
Keyword(s):  
Algal Cell ◽  
Principal Component ◽  
External Perturbation ◽  
Time Lapse ◽  
Chemical Imaging ◽  
Hyperspectral Data ◽  
Aqueous Environment ◽  
Ir Absorption ◽  
Imaging Data ◽  
Mid Infrared

AbstractAlgae are the main primary producers in aquatic environments and therefore of fundamental importance for the global ecosystem. Mid-infrared (IR) microspectroscopy is a non-invasive tool that allows in principle studying chemical composition on a single-cell level. For a long time, however, mid-infrared (IR) imaging of living algal cells in an aqueous environment has been a challenge due to the strong IR absorption of water. In this study, we employed multi-beam synchrotron radiation to measure time-resolved IR hyperspectral images of individual Thalassiosira weissflogii cells in water in the course of acclimation to an abrupt change of CO2 availability (from 390 to 5000 ppm and vice versa) over 75 min. We used a previously developed algorithm to correct sinusoidal interference fringes from IR hyperspectral imaging data. After preprocessing and fringe correction of the hyperspectral data, principal component analysis (PCA) was performed to assess the spatial distribution of organic pools within the algal cells. Through the analysis of 200,000 spectra, we were able to identify compositional modifications associated with CO2 treatment. PCA revealed changes in the carbohydrate pool (1200–950 cm$$^{-1}$$ - 1 ), lipids (1740, 2852, 2922 cm$$^{-1}$$ - 1 ), and nucleic acid (1160 and 1201 cm$$^{-1}$$ - 1 ) as the major response of exposure to elevated CO2 concentrations. Our results show a local metabolism response to this external perturbation.

scholarly journals Advanced Imaging Techniques for Radiotherapy Planning of Gliomas

Cancers ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 1063
Author(s):  
Antonella Castellano ◽  
Michele Bailo ◽  
Francesco Cicone ◽  
Luciano Carideo ◽  
Natale Quartuccio ◽  
...  
Keyword(s):  
Radiation Treatment ◽  
Imaging Modality ◽  
Imaging Techniques ◽  
Clinical Results ◽  
Target Volume ◽  
Radiotherapy Planning ◽  
Tumor Control ◽  
Current Standard ◽  
Volume Delineation ◽  
Hemodynamic Information

The accuracy of target delineation in radiation treatment (RT) planning of cerebral gliomas is crucial to achieve high tumor control, while minimizing treatment-related toxicity. Conventional magnetic resonance imaging (MRI), including contrast-enhanced T1-weighted and fluid-attenuated inversion recovery (FLAIR) sequences, represents the current standard imaging modality for target volume delineation of gliomas. However, conventional sequences have limited capability to discriminate treatment-related changes from viable tumors, owing to the low specificity of increased blood-brain barrier permeability and peritumoral edema. Advanced physiology-based MRI techniques, such as MR spectroscopy, diffusion MRI and perfusion MRI, have been developed for the biological characterization of gliomas and may circumvent these limitations, providing additional metabolic, structural, and hemodynamic information for treatment planning and monitoring. Radionuclide imaging techniques, such as positron emission tomography (PET) with amino acid radiopharmaceuticals, are also increasingly used in the workup of primary brain tumors, and their integration in RT planning is being evaluated in specialized centers. This review focuses on the basic principles and clinical results of advanced MRI and PET imaging techniques that have promise as a complement to RT planning of gliomas.

scholarly journals Multi-pronged approach to human mesenchymal stromal cells senescence quantification with a focus on label-free methods

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Weichao Zhai ◽  
Jerome Tan ◽  
Tobias Russell ◽  
Sixun Chen ◽  
Dennis McGonagle ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Preclinical Model ◽  
Label Free ◽  
Current Standard ◽  
Differentiation Potential ◽  
Human Mesenchymal Stromal Cells ◽  
Endogenous Fluorophores

AbstractHuman mesenchymal stromal cells (hMSCs) have demonstrated, in various preclinical settings, consistent ability in promoting tissue healing and improving outcomes in animal disease models. However, translation from the preclinical model into clinical practice has proven to be considerably more difficult. One key challenge being the inability to perform in situ assessment of the hMSCs in continuous culture, where the accumulation of the senescent cells impairs the culture’s viability, differentiation potential and ultimately leads to reduced therapeutic efficacies. Histochemical $$\upbeta $$ β -galactosidase staining is the current standard for measuring hMSC senescence, but this method is destructive and not label-free. In this study, we have investigated alternatives in quantification of hMSCs senescence, which included flow cytometry methods that are based on a combination of cell size measurements and fluorescence detection of SA-$$\upbeta $$ β -galactosidase activity using the fluorogenic substrate, C$${_{12}}$$ 12 FDG; and autofluorescence methods that measure fluorescence output from endogenous fluorophores including lipopigments. For identification of senescent cells in the hMSC batches produced, the non-destructive and label-free methods could be a better way forward as they involve minimum manipulations of the cells of interest, increasing the final output of the therapeutic-grade hMSC cultures. In this work, we have grown hMSC cultures over a period of 7 months and compared early and senescent hMSC passages using the advanced flow cytometry and autofluorescence methods, which were benchmarked with the current standard in $$\upbeta $$ β -galactosidase staining. Both the advanced methods demonstrated statistically significant values, (r = 0.76, p $$\le $$ ≤ 0.001 for the fluorogenic C$${_{12}}$$ 12 FDG method, and r = 0.72, p $$\le $$ ≤ 0.05 for the forward scatter method), and good fold difference ranges (1.120–4.436 for total autofluorescence mean and 1.082–6.362 for lipopigment autofluorescence mean) between early and senescent passage hMSCs. Our autofluroescence imaging and spectra decomposition platform offers additional benefit in label-free characterisation of senescent hMSC cells and could be further developed for adoption for future in situ cellular senescence evaluation by the cell manufacturers.

scholarly journals Recovery of photoacoustic images based on accurate ultrasound positioning

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Yinhao Pan ◽  
Ningbo Chen ◽  
Liangjian Liu ◽  
Chengbo Liu ◽  
Zhiqiang Xu ◽  
...  
Keyword(s):  
Biological Tissue ◽  
Large Field ◽  
Photoacoustic Microscopy ◽  
Label Free ◽  
Imaging Data ◽  
Motor Speed ◽  
Image Correction ◽  
Microscopy Imaging ◽  
Scanning Method

AbstractPhotoacoustic microscopy is an in vivo imaging technology based on the photoacoustic effect. It is widely used in various biomedical studies because it can provide high-resolution images while being label-free, safe, and harmless to biological tissue. Polygon-scanning is an effective scanning method in photoacoustic microscopy that can realize fast imaging of biological tissue with a large field of view. However, in polygon-scanning, fluctuations of the rotating motor speed and the geometric error of the rotating mirror cause image distortions, which seriously affect the photoacoustic-microscopy imaging quality. To improve the image quality of photoacoustic microscopy using polygon-scanning, an image correction method is proposed based on accurate ultrasound positioning. In this method, the photoacoustic and ultrasound imaging data of the sample are simultaneously obtained, and the angle information of each mirror used in the polygon-scanning is extracted from the ultrasonic data to correct the photoacoustic images. Experimental results show that the proposed method can significantly reduce image distortions in photoacoustic microscopy, with the image dislocation offset decreasing from 24.774 to 10.365 μm.

scholarly journals Anti-neoplastic Potential of Flavonoids and Polysaccharide Phytochemicals in Glioblastoma

Molecules ◽  
2020 ◽  
Vol 25 (21) ◽  
pp. 4895
Author(s):  
Ayesha Atiq ◽  
Ishwar Parhar
Keyword(s):  
Cancer Progression ◽  
Therapeutic Option ◽  
Normal Brain ◽  
Current Standard ◽  
Pharmacological Agents ◽  
Initiation Phase ◽  
Therapeutic Outcomes ◽  
Effective Therapeutic Option ◽  
Patient Prognosis

Clinically, gliomas are classified into four grades, with grade IV glioblastoma multiforme being the most malignant and deadly, which accounts for 50% of all gliomas. Characteristically, glioblastoma involves the aggressive proliferation of cells and invasion of normal brain tissue, outcomes as poor patient prognosis. With the current standard therapy of glioblastoma; surgical resection and radiotherapy followed by adjuvant chemotherapy with temozolomide, it remains fatal, because of the development of drug resistance, tumor recurrence, and metastasis. Therefore, the need for the effective therapeutic option for glioblastoma remains elusive. Previous studies have demonstrated the chemopreventive role of naturally occurring pharmacological agents through preventing or reversing the initiation phase of carcinogenesis or arresting the cancer progression phase. In this review, we discuss the role of natural phytochemicals in the amelioration of glioblastoma, with the aim to improve therapeutic outcomes, and minimize the adverse side effects to improve patient’s prognosis and enhancing their quality of life.

Managing the Edentulous Mandible using Recent Technological Developments: A Case Study

2013 ◽  
Vol 2 (2) ◽  
pp. 50-54
Author(s):  
Ashok Sethi ◽  
Thomas Kaus ◽  
Naresh Sharma ◽  
Peter Sochor
Keyword(s):  
Computed Tomography ◽  
Spatial Information ◽  
Imaging Techniques ◽  
Accurate Information ◽  
Imaging Data ◽  
Surgical Guide ◽  
Prosthetic Reconstruction ◽  
Anterior Teeth ◽  
Edentulous Mandible ◽  
Cad Cam

Safe clinical practice in implant dentistry requires an accurate investigation of the availability of bone for implant placement and the avoidance of critical anatomical structures. Modern imaging techniques using computed tomography (CT) and cone beam computed tomography (CBCT) provide the clinician with the required information. The imaging thus obtained provides accurate representation of the height, width and length of the available bone.1 In addition, whenever adequate radiation dose is used, accurate information about the bone density in Hounsfield units can be obtained. Important spatial information regarding the orientation of the ridges and the relationship to the proposed prosthetic reconstruction can be obtained with the aid of radiopaque templates during the acquisition of CT scan data. Modern software also provides the facility to decide interactively upon the positioning of the implants and is able to relate this to a stereolithographic model constructed from the imaging data. A surgical guide for the accurate positioning of the implants can be constructed. The construction of screw retained prostheses is fraught with difficulties regarding the accuracy of the construction. Accurate fit of the prosthesis is difficult to obtain due to the inherent errors in impression taking, component discrepancies, investing and casting inaccuracies.2,3 CAD/CAM technology eliminates the inaccuracies involved with the investing and casting of superstructures. Clinical Case This case describes the management of an 84 year old female patient, who had recently lost her remaining mandibular anterior teeth. This resulted in the patient's inability to wear conventional dentures in the mandible.

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