Article
Chemistry, Multidisciplinary
Xiaodan Gou, Yiwen Zhang, Zejing Xing, Cheng Ma, Changjie Mao, Jun-Jie Zhu
Summary: In this study, a site-selective heat boosting electrochemiluminescence microscopy technique was reported. By generating a micron-scale heat point in situ at the electrode-solution interface, a 63-fold enhancement in luminescence intensity and a 0.2 V advance in applied voltage were achieved. Experimental results and finite element simulation showed that the fundamental reasons for this phenomenon were accelerated reaction rate and thermal convection via a photothermal effect. The concentrated electrochemiluminescence not only boosted the contrast of single cells by 20.54 times but also enabled the site-selective cell-by-cell analysis of the heterogeneous membrane protein abundance. This electrochemical visualization method has great potential in the highly sensitive and selective analysis of local electron transfer events.
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
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)
Review
Chemistry, Analytical
Zrinko Baricevic, Zahra Ayar, Samuel M. Leitao, Miranda Mladinic, Georg E. Fantner, Jelena Ban
Summary: Time-lapse light microscopy combined with in vitro neuronal cultures has made a significant contribution to Developmental Neuroscience. Although fluorescently labeled tags and dyes have provided advancements in cell imaging, long-term recordings remain challenging due to phototoxicity and interference with cell physiology. Hence, label-free high-resolution methods, such as scanning ion conductance microscopy (SICM) or digital holography microscopy (DHM) are proposed for studying live cell dynamics during neuronal development and regeneration.
Article
Cell Biology
Joanna W. Pylvanainen, Estibaliz Gomez-de-Mariscal, Ricardo Henriques, Guillaume Jacquemet
Summary: Live imaging, particularly when combined with fluorescence microscopy, is a powerful tool for observing cellular components in real time. However, implementing live imaging and analyzing the resulting datasets is challenging due to issues such as drift, phototoxicity, and dataset size. Recent advances in bioimage analysis, including deep learning, are revolutionizing how live imaging is performed. Important computational methods for live imaging include drift correction, denoising, super-resolution imaging, artificial labeling, tracking, and time series analysis. Self-driving microscopy has also seen recent advancements.
CURRENT OPINION IN CELL BIOLOGY
(2023)
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
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)
Review
Optics
Herbert Schneckenburger, Verena Richter
Summary: This short overview discusses 3D live cell imaging, relevant samples, challenges like optical sectioning and phototoxicity, and enhanced microscopy methods with some examples and applications. Future advancements may involve increasing microscopic resolution and utilizing molecular sensing techniques.
Article
Multidisciplinary Sciences
Rajesh Ranjan, Xin Chen
Summary: This study presents a method for conducting super-resolution timelapse fluorescence live cell imaging in situ, compatible with low fluorescent intensity cells and capable of visualizing multiple subcellular structures. Special procedures in sample preparation and immobilizing specimens are necessary for this technique, but it is broadly applicable to various cell types.
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
(2021)
Review
Plant Sciences
Maria Ada Prusicki, Martina Balboni, Kostika Sofroni, Yuki Hamamura, Arp Schnittger
Summary: Live-cell imaging is a powerful tool for understanding cellular processes, especially in meiosis where chromosomes and cellular components exhibit complex dynamics over a short period of time. Recent advances in live-cell imaging now allow real-time analysis of meiotic events in plants.
FRONTIERS IN PLANT SCIENCE
(2021)
Article
Multidisciplinary Sciences
Hela Benaissa, Karim Ounoughi, Isabelle Aujard, Evelyne Fischer, Rosette Goiame, Julie Nguyen, Alison G. Tebo, Chenge Li, Thomas Le Saux, Giulia Bertolin, Marc Tramier, Lydia Danglot, Nicolas Pietrancosta, Xavier Morin, Ludovic Jullien, Arnaud Gautier
Summary: Fluorescent reporters covering the visible spectrum are essential for imaging living cells and organisms. The authors have developed a collection of fluorogenic chromophores spanning from blue to red using a single engineered protein tag. This tool allows for tuning fluorescence color and properties, providing versatility for imaging proteins in live cells and optimizing FRET biosensors.
NATURE COMMUNICATIONS
(2021)
Article
Chemistry, Multidisciplinary
Georgina C. Gavins, Katharina Groeger, Michael D. Bartoschek, Philipp Wolf, Annette G. Beck-Sickinger, Sebastian Bultmann, Oliver Seitz
Summary: DNA nanotechnology offers opportunities for manipulating and imaging proteins on cell surfaces by creating nucleic acid-protein junctions using PNA tags. This method allows for quick assembly of different fluorescent dyes, increased brightness, reversible labeling, and is applicable to various cell and protein systems.
Article
Biochemical Research Methods
Christoffer Edlund, Timothy R. Jackson, Nabeel Khalid, Nicola Bevan, Timothy Dale, Andreas Dengel, Sheraz Ahmed, Johan Trygg, Rickard Sjoegren
Summary: The LIVECell dataset consists of annotated phase-contrast images of over 1.6 million cells, aiming to improve the training of image segmentation models through deep learning. Its creation and utilization help explore biological phenomena and conduct high-throughput quantitative imaging studies.
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
Chemistry, Analytical
Jia Zhang, Xinwei Gao, Luwei Wang, Yong Guo, Yinru Zhu, Zhigang Yang, Wei Yan, Junle Qu
Summary: STED microscopy is a super-resolution imaging technology that allows studying dynamic processes of live cells at the nanometer scale. Researchers developed a low-power two-color STED microscope for live cell imaging, showing great potential for long-term dynamic imaging of live cells.
Article
Chemistry, Multidisciplinary
Congnyu Che, Ruiyang Xue, Nantao Li, Prashant Gupta, Xiaojing Wang, Bin Zhao, Srikanth Singamaneni, Shuming Nie, Brian T. Cunningham
Summary: This study presents a method using magneto-plasmonic nanoparticles to accelerate single-molecule sensing for quantification of miRNA. The method has a fast response time, low detection limit, broad dynamic range, and good single-base mismatch selectivity, making it suitable for minimally invasive biomarker quantification.
Article
Chemistry, Multidisciplinary
Nantao Li, Xiaojing Wang, Joseph Tibbs, Congnyu Che, Ana Sol Peinetti, Bin Zhao, Leyang Liu, Priyash Barya, Laura Cooper, Lijun Rong, Xing Wang, Yi Lu, Brian T. Cunningham
Summary: This study introduces a label-free biosensing method for quickly detecting and quantifying intact viruses in human saliva with high selectivity and sensitivity.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(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
Chemistry, Multidisciplinary
Xiaojing Wang, Skye Shepherd, Nantao Li, Congnyu Che, Tingjie Song, Yanyu Xiong, Isabella Rose Palm, Bin Zhao, Manish Kohli, Utkan Demirci, Yi Lu, Brian T. T. Cunningham
Summary: In this study, an efficient target recycling amplification process (TRAP) using photonic resonator absorption microscopy was developed for the digital detection of exosomal microRNAs (miRNAs). The TRAP approach demonstrated high sensitivity, robust selectivity, and a broad dynamic range, making it ideal for quantifying miRNA biomarkers in low concentrations or sample volume. Compared to traditional qRT-PCR, TRAP showed similar accuracy in profiling exosomal miRNAs derived from cancer cells, but with significantly enhanced detection limits for miRNA-375 and miRNA-21.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
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
Biophysics
Leyang Liu, Joseph Tibbs, Nantao Li, Amanda Bacon, Skye Shepherd, Hankeun Lee, Neha Chauhan, Utkan Demirci, Xing Wang, Brian T. Cunningham
Summary: In this paper, a compact Photonic Resonator Interferometric Scattering Microscope (PRISM) designed for point-of-use environments and applications is presented. It allows label-free detection and digital counting of nanometer-scaled objects such as nanoparticles, viruses, extracellular vesicles, and protein molecules. The instrument incorporates two innovative elements to facilitate operation in ordinary laboratory environments and achieves stable image contrast over time and spatial position through an automated focusing module.
BIOSENSORS & BIOELECTRONICS
(2023)
Article
Chemistry, Multidisciplinary
Priyash Barya, Yanyu Xiong, Skye Shepherd, Rohit Gupta, Lucas D. Akin, Joseph Tibbs, Hankeun Lee, Srikanth Singamaneni, Brian T. Cunningham
Summary: Assays using fluorophores are common in life science research and diagnostics. However, weak emission intensity limits the detection limits, requiring a large number of labeled target molecules to improve signal-to-noise ratio. This study demonstrates that the synergistic coupling of plasmonic and photonic modes can significantly enhance the emission from fluorophores, resulting in a 52-fold improvement in signal intensity. The method is successfully applied to a sandwich immunoassay for human interleukin-6, achieving a detection limit three orders of magnitude lower than standard immunoassays.
Review
Chemistry, Analytical
Yanyu Xiong, Skye Shepherd, Joseph Tibbs, Amanda Bacon, Weinan Liu, Lucas D. Akin, Takhmina Ayupova, Seemesh Bhaskar, Brian T. Cunningham
Summary: Nanoscale fluorescence emitters are efficient in measuring biomolecular interactions but limited in applications requiring single-unit observations due to various challenges. Photonic crystal structures offer solutions for fluorescence enhancement by optimizing numerical aperture objectives, reducing fluorescence intermittency, and improving photon collection efficiency. This review provides an overview of photonic crystals, their design, fabrication, and sensing principles, as well as recent applications in biosensors for nucleic acids, proteins, and steroids. The review also discusses current challenges and future prospects of fluorescence enhancement with photonic-plasmonics coupling in point-of-care biosensing and monitoring analytes of biological and environmental relevance.
Review
Chemistry, Analytical
Amanda Bacon, Weijing Wang, Hankeun Lee, Saurabh Umrao, Prima Dewi Sinawang, Demir Akin, Kodchakorn Khemtonglang, Anqi Tan, Sabina Hirshfield, Utkan Demirci, Xing Wang, Brian T. Cunningham
Summary: The ability for individuals to self-test for HIV is crucial in preventing transmission, especially when combined with other biomedical prevention methods like PrEP. In this article, we review recent advancements in HIV self-testing and sampling methods, and explore the potential impact of new materials and techniques developed for more effective point-of-care SARS-CoV-2 diagnostics. We discuss the limitations of current HIV self-testing technologies and the need for improvements in sensitivity, speed, simplicity, and cost to enhance diagnostic accuracy and accessibility. We also examine potential paths towards the next generation of HIV self-testing through sample collection materials, biosensing assays, and miniaturized instruments, as well as the implications for self-monitoring HIV viral load and other infectious diseases.
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)
Article
Chemistry, Analytical
Aaron M. Jankelow, Hankeun Lee, Weijing Wang, Trung-Hieu Hoang, Amanda Bacon, Fu Sun, Seol Chae, Victoria Kindratenko, Katherine Koprowski, Robert A. Stavins, Dylann D. Ceriani, Zachary W. Engelder, William P. King, Minh N. Do, Rashid Bashir, Enrique Valera, Brian T. Cunningham
Summary: In this study, we report an integrated system for rapid detection of viral pathogens. The system can perform sample processing and nucleic acid amplification in 20 minutes and can analyze the results using a smartphone. The system offers the advantages of low cost, high accuracy, and simplicity in operation while achieving rapid detection.
