Article
Biotechnology & Applied Microbiology
Andreas Boden, Francesca Pennacchietti, Giovanna Coceano, Martina Damenti, Michael Ratz, Ilaria Testa
Summary: A new 3D pRESOLFT microscope method with sub-80-nm resolution has been developed to visualize the volumetric architecture of organelles and molecules inside whole living cells, allowing for targeted 3D confinement of fluorescence and observation of dynamic structural alterations in cells.
NATURE BIOTECHNOLOGY
(2021)
Article
Biology
Jeremy Copperman, Sean M. Gross, Young Hwan Chang, Laura M. Heiser, Daniel M. Zuckerman
Summary: Time-lapse imaging is a powerful tool to study cell dynamics, but analyzing morphological changes over time is challenging. In this study, we use trajectory embedding to analyze cellular behavior based on multiple time points simultaneously. We apply this method to study the effects of microenvironmental perturbagens on MCF10A mammary epithelial cells. Our analysis reveals ligand-specific regulation of cell state transitions and provides descriptive models of single-cell trajectories. Incorporating trajectories improves the characterization of cell states and enables better identification of phenotypes compared to snapshot-based analysis. This approach has broad applications in live-cell imaging for studying cell responses.
COMMUNICATIONS BIOLOGY
(2023)
Article
Multidisciplinary Sciences
Alice Sandmeyer, Lili Wang, Wolfgang Huebner, Marcel Mueller, Benjamin K. Chen, Thomas Huser
Summary: Cost-effective, highly portable, and easy to use high-resolution live-cell imaging systems can revolutionize research in challenging environments, enabling real-time tracking of virus particles and studying infection mechanisms.
Article
Multidisciplinary Sciences
Chenfei Hu, Shenghua He, Young Jae Lee, Yuchen He, Edward M. Kong, Hua Li, Mark A. Anastasio, Gabriel Popescu
Summary: The study introduces a method for assessing cell viability without the need for labeling, utilizing deep learning techniques combined with quantitative phase imaging to accurately determine cell survival status without causing damage to the cells.
NATURE COMMUNICATIONS
(2022)
Article
Biochemistry & Molecular Biology
Anayat Bhat, Shuang Li, Daniel Hammler, Martin J. Winterhalder, Andreas Marx, Andreas Zumbusch
Summary: The hydrolysis of nucleotides is important for cellular energy, and the transfer of phosphates to proteins is crucial for post-translational modification. A novel fluorogenic adenosine 5'-tetraphosphate (Ap4) analog has been developed for real-time monitoring of ATP hydrolysis within live cells, with potential applications in studying cellular processes involving Ap4 hydrolysis.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Biology
Anthony Santella, Irina Kolotuev, Caroline Kizilyaprak, Zhirong Bao
Summary: Analyzing brain development, structure, and function across imaging modalities can provide valuable information. This study combines electron microscopy and time-lapse fluorescence microscopy to study the emergence of the nervous system in Caenorhabditis elegans embryogenesis.
Article
Chemistry, Multidisciplinary
Tom Baladi, Jesper R. Nilsson, Audrey Gallud, Emanuele Celauro, Cecile Gasse, Fabienne Levi-Acobas, Ivo Sarac, Marcel R. Hollenstein, Anders Dahlen, Elin K. Esbjorner, L. Marcus Wilhelmsson
Summary: The study introduces a stealth labeling strategy to efficiently generate RNA transcripts and translate them into correct proteins, providing an important tool for biological research and RNA delivery studies.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2021)
Article
Materials Science, Multidisciplinary
Franziska Weigl, Carina Blum, Ana Sancho, Juergen Groll
Summary: In recent decades, hybrid characterization systems have become important in cellular biomechanics research. This paper introduces a multifunctional device that combines Fluorescence Microscopy (FL) with FluidFM technology, allowing for real-time optical tracking of cell detachment processes while acquiring force-distance curves simultaneously.
ADVANCED MATERIALS TECHNOLOGIES
(2022)
Article
Chemistry, Analytical
Hong Wang, Guangmei Han, Hesen Tang, Ruilong Zhang, Zhengjie Liu, Yingqiang Sun, Bianhua Liu, Junlong Geng, Zhongping Zhang
Summary: Fluorescence microscopy is an important tool in cell biology research, but its use for long-term and strong-illumination imaging of live cells is limited by photobleaching and phototoxicity. This study introduces a new concept of fluorophore engineering chemistry, using synchronous photoactivation-imaging (SPI) fluorophores to overcome these limitations. The SPI fluorophores allow continuous replenishing of bright-state probes and reduce photocytotoxicity, enabling long-term imaging of live cells under intensive illumination.
ANALYTICAL CHEMISTRY
(2023)
Article
Chemistry, Multidisciplinary
Martin Priessner, Peter A. Summers, Benjamin W. Lewis, Magdalena Sastre, Liming Ying, Marina K. Kuimova, Ramon Vilar
Summary: A new optical probe based on BODIPY has been developed, which exhibits fluorescence intensity switch-on upon binding to copper(I) and can be used to visualize copper(I) pools in lysosomes of live cells.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Article
Cell Biology
Amy H. Ponsford, Thomas A. Ryan, Andrea Raimondi, Emanuele Cocucci, Susanne A. Wycislo, Florian Froehlich, Laura E. Swan, Massimiliano Stagi
Summary: Peripheral lysosomes are not less acidic compared to perinuclear lysosomes, and lysosomal pH has a certain impact on the transmission of lysosomal stress signals.
Article
Biochemical Research Methods
Arindam Ghosh, Alexey Chizhik, Narain Karedla, Joerg Enderlein
Summary: Super-resolution fluorescence imaging is commonly used to study the spatial organization of biological structures at molecular scales, achieving resolutions beyond the classical optical limit. Various methods have been developed for super-resolution imaging along the optical axis, with some reaching nanometer accuracy in 3D localization. However, these approaches can be technically complex and have limited applicability. Metal-induced energy transfer imaging offers a simpler alternative for nanometer-scale axial localization of fluorophores.
Article
Chemistry, Analytical
Su Liang, Zeyu Wang, Luyu Qi, Chao Tang, Yiming Zhang, Qun Luo, Yayun Wu, Jinghe Yuan, Yao Zhao, Yanyan Zhang, Xiaohong Fang, Shijun Wang, Fuyi Wang
Summary: This study designed a fluorescent probe, ATTO565-WGN, which showed similar anti-proliferative activity to wogonin against human cancer cell lines and demonstrated mitochondrial localization through confocal fluorescence imaging. The conjugated ATTO565 fluorophore enabled STED fluorescence living cell imaging with ultrahigh spatial resolution, facilitating the investigation of wogonin-biological target interactions in living cells at mitochondria. This work provides novel insights into rational design of mitochondrion targeting fluorescence probes for ultrahigh resolution living cell imaging.
Article
Optics
Zewei Luo, Ge Wu, Mengting Kong, Zhi Chen, Zhengfei Zhuang, Junchao Fan, Tongsheng Chen
Summary: Forster resonance energy transfer (FRET) microscopy is a valuable tool for studying biological systems, but super-resolution FRET imaging in living cells is challenging. In this study, we propose a SR FRET method called SIM-FRET that combines SR structured illumination microscopy (SIM) imaging and acceptor sensitized emission FRET imaging to achieve live-cell quantitative SR FRET imaging. Our method provides enhanced spatial resolution and maintains the advantages of quantitative FRET analysis. We validate the effectiveness of SIM-FRET through simulated models and live-cell samples, and demonstrate its ability to reveal the intricate structure of FRET signals.
PHOTONICS RESEARCH
(2023)
Article
Chemistry, Analytical
Ian R. D. Johnson, Elley E. Rudebeck, Martin J. Sweetman, Alexandra Sorvina, Trent D. Ashton, Frederick M. Pfeffer, Douglas A. Brooks, Shane M. Hickey
Summary: Lipid droplets (LDs) are dynamic organelles that not only serve as lipid reservoirs, but also play important roles in cell metabolism and growth. The study of LDs and their interactions with other cellular compartments provides new insights into normal cell biology and disease processes. In this study, two newly synthesized compounds were evaluated as selective LD stains, showing impressive versatility in staining LDs in various cellular contexts. These new imaging agents have the potential to further unravel the complex biology of LDs and their role in disease states.
SENSORS AND ACTUATORS B-CHEMICAL
(2022)
Article
Engineering, Biomedical
Victoria Barnhouse, Nathan Petrikas, Cody Crosby, Janet Zoldan, Brendan Harley
Summary: The study developed and characterized a three-dimensional perivascular tissue model to investigate the influence of the perivascular secretome on HSC behavior, finding that perivascular conditioned media promoted maintenance of a greater fraction of hematopoietic stem and progenitor cells within a 4-day culture period.
ANNALS OF BIOMEDICAL ENGINEERING
(2021)
Article
Engineering, Biomedical
Marley J. Dewey, Andrey Nosatov, Kiran Subedi, Ramille Shah, Adam Jakus, Brendan A. C. Harley
Summary: Incorporating 3D-printed polymer or ceramic-based mesh into mineralized collagen scaffolds improves mechanical and biological activity, supporting mesenchymal stem cell osteogenesis and new bone formation. The mesh can instruct cell processes that aid osteogenicity, specifically inhibiting osteoclast-mediated bone resorption through increased secretion of osteoprotegerin, an endogenous inhibitor of osteoclast activity.
ACTA BIOMATERIALIA
(2021)
Article
Multidisciplinary Sciences
N. R. Richbourg, M. Wancura, A. E. Gilchrist, S. Toubbeh, B. A. C. Harley, E. Cosgriff-Hernandez, N. A. Peppas
Summary: This study investigates how changes in synthesis conditions affect the network structure of hydrogels, and identifies linear correlations between synthesis variables and swelling behavior. It also demonstrates that complex biopolymer structures can disrupt the linear synthesis-swelling correlations. These findings provide insight into controlling hydrogel physical properties through structural design and can be used to optimize biomedically relevant hydrogels.
Article
Chemistry, Multidisciplinary
Mai T. Ngo, Victoria R. Barnhouse, Aidan E. Gilchrist, Bhushan P. Mahadik, Christine J. Hunter, Joy N. Hensold, Nathan Petrikas, Brendan A. C. Harley
Summary: Biomaterials mimicking microenvironmental signals from the stem cell niche could improve platforms regulating stem cell behavior. Vascular-derived signals offer an important alternative signaling axis, but dose-dependent relationships between angiocrine signals and stem cell fate are difficult to determine in animal models and 2D cell cultures. This study uses microfluidic devices to create 3D hydrogels with lateral gradients in vascular density, revealing spatial patterns of HSC phenotype in response to angiocrine signals and showcasing the influence of local angiocrine signals on HSC fate.
ADVANCED FUNCTIONAL MATERIALS
(2021)
Article
Neurosciences
Edward R. Neves, Brendan A. C. Harley, Sara Pedron
Summary: Brain tumors lack effective treatments, and understanding tumor progression and resistance mechanisms is unclear. Technological advances in in vitro modeling are being used to increase the efficiency of preclinical systems. Microphysiological brain tumor models may offer insights into disease mechanisms and predict patient response to anticancer treatments.
BRAIN RESEARCH BULLETIN
(2021)
Article
Cell & Tissue Engineering
Samantha G. Zambuto, Shemona Rattila, Gabriela Dveksler, Brendan A. C. Harley
Summary: This study investigates the effects of PSG1 and PSG9, along with epidermal growth factor and Nodal, on trophoblast motility using 3D trophoblast motility assays. The results show that PSG9 reduces trophoblast motility while PSG1 increases motility, highlighting the importance of these factors in regulating trophoblast response during early pregnancy.
CELLULAR AND MOLECULAR BIOENGINEERING
(2022)
Review
Engineering, Biomedical
Aidan E. Gilchrist, Brendan A. C. Harley
Summary: This review describes recent advances in biomaterials systems to replicate features of the hematopoietic niche, providing stimuli and signals to support the encapsulation and expansion of hematopoietic stem cells. These engineered systems enable the study of the dynamic nature of matrix and cell-cell signaling.
ADVANCED HEALTHCARE MATERIALS
(2022)
Review
Engineering, Biomedical
Aleczandria S. Tiffany, Brendan A. C. Harley
Summary: This review provides an overview of growth plates, their role in skeletal development, injuries and diseases associated with growth plates, and the opportunities and challenges for developing biomaterial models for studying growth plate biology and diseases. It also describes the potential of sophisticated in vitro biomaterial models for studying growth plate remodeling and developing tissue engineering applications.
ADVANCED HEALTHCARE MATERIALS
(2022)
Article
Chemistry, Analytical
Isamar Pastrana-Otero, Sayani Majumdar, Aidan E. Gilchrist, Brendan A. C. Harley, Mary L. Kraft
Summary: This study demonstrates that a combination of Raman microspectroscopy and partial least squares discriminant analysis (PLS-DA) can be used to location-specifically identify individual living cells from the six most immature hematopoietic cell populations. By utilizing spectral features associated with cell biochemistries, the differentiation stages of cells can be accurately identified.
ANALYTICAL CHEMISTRY
(2022)
Article
Materials Science, Biomaterials
Samantha G. Zambuto, Ishita Jain, Kathryn B. H. Clancy, Gregory H. Underhill, Brendan A. C. Harley
Summary: The endometrium undergoes significant changes in tissue architecture and composition during the menstrual cycle and pregnancy. The remodeling of the endometrial extracellular matrix (ECM) plays a crucial role in maintaining endometrial homeostasis. The endometrial basement membrane, which separates the endometrial epithelium from the underlying ECM, is an important component of the tissue microenvironment. This study focuses on investigating the attachment and expression of cytokeratin 18 (CK18) in endometrial epithelial cells (EEC) in response to different combinations of basement membrane proteins. The findings provide insights into tissue-engineered models of the endometrial microenvironment.
ACS BIOMATERIALS SCIENCE & ENGINEERING
(2022)
Article
Chemistry, Multidisciplinary
Mai T. Ngo, Jann N. Sarkaria, Brendan A. C. Harley
Summary: This article introduces a 3D in vitro model of the perivascular niche, which helps to understand the behavior and progression of brain tumor cells around blood vessels, and reveals the role of pericytes and astrocytes in regulating tumor cell invasion, proliferation, and therapeutic response.
Article
Engineering, Biomedical
Marley J. Dewey, Alan J. Collins, Aleczandria Tiffany, Victoria R. Barnhouse, Crislyn Lu, Vasiliki Kolliopoulos, Isha Mutrejag, Noreen J. Hickokh, Brendan A. C. Harley
Summary: The design of biomaterials for bone regeneration needs to reduce bacterial attachment and biofilm formation to prevent infection. This study investigates the ability of mineralized collagen biomaterials to resist infection and examines the effect of manuka honey on bacterial colonization, stem cell osteogenesis, and vasculature formation.
Article
Engineering, Biomedical
Marley J. Dewey, Kyle B. Timmer, Ashley Blystone, Crislyn Lu, Brendan A. C. Harley
Summary: Current treatments for craniomaxillofacial (CMF) defects require the design of instructive biomaterials. This study shows that the incorporation of ascorbic acid into a mineralized collagen scaffold can promote osteogenesis of human mesenchymal stem cells (hMSCs) and influence gene expression related to bone formation. These findings highlight the importance of ascorbic acid in the design of mineralized collagen scaffolds for improved CMF outcomes.
JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART A
(2023)
Article
Materials Science, Biomaterials
Vasiliki Kolliopoulos, Maxwell Polanek, Hui Xu, Brendan Harley
Summary: This study investigates the influence of the initial inflammatory stance of human mesenchymal stem cells (hMSCs) on key osteogenic, angiogenic, and immuno-modulatory criteria in a class of mineralized collagen scaffolds for craniomaxillofacial (CMF) bone repair. The results highlight the importance of scaffold properties on the sustained kinetics of cell response to an inflammatory stimulus.
ACS BIOMATERIALS SCIENCE & ENGINEERING
(2023)
Review
Chemistry, Multidisciplinary
Marley J. Dewey, Brendan A. C. Harley
Summary: The bone microenvironment is complex, requiring biomaterial design to overcome obstacles such as infection, fibrous tissue formation, non-bone tissue formation, and voids in bone infill. Novel biomaterial designs may provide approaches to effectively induce osteogenesis and new bone formation while preventing failure due to unaddressed issues.