Article
Optics
Kevin Affannoukoue, Simon Labouesse, Guillaume Maire, Laurent Gallais, Julien Savatier, Marc Allain, Md Rasedujjaman, Loic Legoff, Jerome Idier, Renaud Poincloux, Florence Pelletier, Christophe Leterrier, Thomas Mangeat, Anne Sentenac
Summary: A benefit of random illumination microscopy (RIM) is that it improves the resolution and linearity of structured illumination microscopy using speckled illumination. In this study, RIM was implemented in the TIRF configuration for imaging biological processes close to the coverslip surface, achieving high-resolution images on fixed samples and dynamic images on live macrophages. The simplicity of the experimental setup and imaging protocol, as well as the robustness of the data processing, make TIRF-RIM a preferred method for super-resolution TIRF imaging.
Article
Optics
Zetao Fan, Yan Kuai, Xi Tang, Yifan Zhang, Douguo Zhang
Summary: This paper proposes a novel TIRF microscope based on a photonic chip, which can replace conventional objectives or prisms for simplified optical configuration and shadowless illumination.
Article
Biology
Sarina Veit, Laura Charlotte Paweletz, Soren S-R Bohr, Anant K. Menon, Nikos S. Hatzakis, Thomas Guenther Pomorski
Summary: Reconstitution of membrane proteins into model membranes is crucial for functional analysis, but existing model-membrane systems have limitations in sample heterogeneity and insufficient knowledge of lipid and protein content. This study presents a new microscopy-based bleaching protocol that allows parallel analysis of multiple parameters of individual proteoliposomes prepared with fluorescently tagged membrane proteins and lipid markers.
Review
Chemistry, Analytical
Louis Colson, Youngeun Kwon, Soobin Nam, Avinashi Bhandari, Nolberto Martinez Maya, Ying Lu, Yongmin Cho
Summary: In this review, we provide an overview of the state-of-the-art advancements in single-molecule fluorescence imaging techniques. We systematically explore the advanced implementations of in vitro single-molecule imaging techniques using total internal reflection fluorescence (TIRF) microscopy and delve into the compatibility of microfluidic technology for single-molecule fluorescence imaging. We also summarize the current challenges and prospects of fluorescence-based single-molecule imaging techniques.
Article
Chemistry, Multidisciplinary
Yannic Kerkhoff, Latifeh Azizi, Vasyl V. Mykuliak, Vesa P. Hytonen, Stephan Block
Summary: This study presents a microfluidics-based single-molecule force spectroscopy method that allows for parallel quantification of hundreds of single-molecule targets. By combining total internal reflection microscopy and microfluidics, the method achieves high spatial resolution and sensitivity. A cluster analysis is implemented to handle the large amount of data generated. The method is validated by studying the mechanical response of polyethylene glycol linkers and the binding strength of biotin-NeutrAvidin complexes.
Article
Multidisciplinary Sciences
Alan M. Szalai, Bruno Siarry, Jeronimo Lukin, David J. Williamson, Nicolas Unsain, Alfredo Caceres, Mauricio Pilo-Pais, Guillermo Acuna, Damian Refojo, Dylan M. Owen, Sabrina Simoncelli, Fernando D. Stefani
Summary: The photometric method SIMPLER allows decoding of the axial position of single molecules in a total internal reflection fluorescence microscope without hardware modification, delivering nearly isotropic nanometric resolution 3D images in single-molecule localization microscopy. Achieving high axial resolution remains challenging in this field.
NATURE COMMUNICATIONS
(2021)
Article
Chemistry, Analytical
Teliang Zhang, Xueliang Wang, Youjun Zeng, Songfeng Huang, Xiaoqi Dai, Weifu Kong, Qian Liu, Jiajie Chen, Junle Qu, Yonghong Shao
Summary: Surface plasmon resonance microscopy (SPRM) is widely used in biological fields for its high spatial resolution and label-free detection. In this study, a home-built SPRM system based on total internal reflection (TIR) was utilized to study the imaging of a single nanoparticle. By using a ring filter and deconvolution algorithm, the parabolic tail of the nanoparticle image was removed, achieving a spatial resolution of 248 nm. Additionally, specific binding between human IgG antigen and goat anti-human IgG antibody was successfully measured using TIR-based SPRM, demonstrating its capability in imaging sparse nanoparticles and monitoring biomolecular interactions.
Review
Endocrinology & Metabolism
Bastian Gaus, Dennis Bruening, Sofie Gross, Michael Mueller, Ingo Rustenbeck
Summary: Before the development of TIRF microscopy, the fate of insulin granules before secretion was inferred through biochemical investigations, electron microscopy, and electrophysiological measurements. The application of TIRF microscopy allowed for a better understanding of insulin granule dynamics in pancreatic beta cells, revealing the relationship between granule fusion and binding to the plasma membrane and cortical actin web. This highlighted the dynamic and heterogeneous nature of submembrane granules in contributing to the metabolic memory of beta cells.
FRONTIERS IN ENDOCRINOLOGY
(2022)
Article
Optics
Daniel Fan, Jelmer Cnossen, Shih-Te Hung, Dimitri Kromm, Nynke H. Dekker, Gerard J. Verbiest, Carlas S. Smith
Summary: In order to better understand the interactions between biological molecules, it is crucial to have a high optical resolution in all three dimensions. However, the inherently lower axial resolution of microscopes poses a limitation in fluorescence imaging, especially in fluorescence-based single molecule localization microscopy (SMLM). In this study, a method is presented to improve the axial localization precision in SMLM by combining point-spread-function engineering with total internal reflection fluorescence (TIRF) fields. This approach allows for the extraction of additional axial location information from the emitted photons, resulting in a two-fold improvement in axial localization compared to TIRF-based SMLM using astigmatic PSFs. Theoretical resolution gains under various imaging conditions are calculated using the Cramer Rao Lower Bound (CRLB), a commonly used metric to compute the best attainable localization precision in SMLM.
OPTICS COMMUNICATIONS
(2023)
Article
Chemistry, Analytical
Jiying Xu, Caixin Huang, Liangju Li, Ying Zhao, Zhenpeng Guo, Yi Chen, Pengfei Zhang
Summary: ESM technology allows for the measurement of ligand interactions with membrane proteins and analysis of cell adhesion properties and cell activity changes, providing a new method for studying cellular processes and drug screening.
Article
Biochemistry & Molecular Biology
Joerg Nikolaus, Kasey Hancock, Maria Tsemperouli, David Baddeley, Erdem Karatekin
Summary: Researchers used a polarization-controlled total internal reflection fluorescence (pTIRF) microscope to monitor the fusion of proteoliposomes with planar lipid bilayers, investigating membrane fusion pore dynamics. By varying excitation polarization angle, they identified a dye-dependent optimal polarization for maximal fluorescence increase upon fusion, aiding event detection and lipid transfer kinetics analysis.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2021)
Article
Biology
Yerdos A. Ordabayev, Larry J. Friedman, Jeff Gelles, Douglas L. Theobald, Ruben L. Gonzalez
Summary: Researchers have developed an unsupervised machine learning method called Tapqir, which incorporates Bayesian probabilistic programming, to automatically analyze CoSMoS data. This method takes into account the uncertainties in image analysis and enables accurate analysis of biochemical reaction mechanisms.
Article
Biochemistry & Molecular Biology
Balint Kiss, Luca Annamaria Kiss, Zsombor David Lohinai, Dorottya Mudra, Hedvig Tordai, Levente Herenyi, Gabriella Csik, Miklos Kellermayer
Summary: This study used TIRF microscopy to uncover the spatial dynamics of target recognition and binding by individual T7 phage particles, and the details of phage anchoring were observed by atomic force microscopy. The results show that viral infection leads to structural changes and lysis of the host cell, and genome release is mechanically controlled. The single-particle approach provides unprecedented insights into the complete viral cycle.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Polymer Science
J. Alejandro Rivera-Moran, Peter R. Lang
Summary: This paper investigates the influence of noise effects, particle size, and sampling time on total internal reflection microscopy (TIRM) measurement results. It finds that detector shot noise and prolonged sampling times may cause erroneous results in steep parts of the interaction potential, while background noise has negligible effects. Noise does not significantly affect dynamic data, but lengthy sampling times and small probe particle radii can cause issues. Additionally, the study observes that dynamic results may differ from standard hydrodynamic predictions due to partial slip.
Article
Chemistry, Analytical
Hongqi Chen, Wanying Xia, Qian Gao, Lun Wang
Summary: A fast and efficient quantitative image analysis method has been developed for the determination of HSO3- concentration, using upconversion nanoparticles and cyanine dye molecules to build a luminescence energy transfer system. The method significantly reduces sample consumption, allows for quick and efficient analysis, and shows a wider linear range, lower detection limit, and higher detection speed compared to the classical luminescence spectrometer.
Review
Food Science & Technology
Georg Sandner, Alice Koenig, Melanie Wallner, Julian Weghuber
Summary: In the field of (food) toxicology, there is a growing trend to replace animal trials with alternative methods to assess adverse health effects in humans. While advancements have been made in in vitro cell culture and in silico modeling, there is a lack of accurate predictions of metabolic actions. Alternative in vivo models are expected to bridge the gap between cell culture and animal trials.
CRITICAL REVIEWS IN FOOD SCIENCE AND NUTRITION
(2022)
Review
Chemistry, Analytical
Mara Heckmann, Gerald Klanert, Georg Sandner, Peter Lanzerstorfer, Manfred Auer, Julian Weghuber
Summary: This article reviews the methods used to quantify the translocation of GLUT4 to the plasma membrane, including microscopy-based techniques and biochemical and spectrometric approaches. These methods are of great importance in researching and identifying substances with insulinomimetic properties.
METHODS AND APPLICATIONS IN FLUORESCENCE
(2022)
Article
Chemistry, Analytical
Tina Karimian, Roland Hager, Andreas Karner, Julian Weghuber, Peter Lanzerstorfer
Summary: This article introduces a simplified activation procedure for depositing biomolecule micropatterns on untreated solid substrates, allowing for direct microcontact printing and strong protein attachment via avidity binding. The specificity and suitability of this method have been demonstrated in proof-of-concept experiments, enhancing the accessibility of protein micropatterning for cell biological research.
Article
Chemistry, Multidisciplinary
Carmen M. Dominguez, Miguel Garcia-Chame, Ulrike Mueller, Andreas Kraus, Klavdiya Gordiyenko, Ahmad Itani, Heiko Haschke, Peter Lanzerstorfer, Kersten S. Rabe, Christof M. Niemeyer
Summary: This article reports on the high dynamic interaction between streptavidin and DNA origami nanostructures, and provides detailed insights into this interaction using a novel high-speed atomic force microscope. The study also demonstrates a correlation between this interaction and cellular activation. This work is significant for further standardizing the use of DNA nanostructures in the study of biological processes.
Article
Biochemistry & Molecular Biology
Mara Heckmann, Nadiia Sadova, Ivana Drotarova, Stefanie Atzmuller, Bettina Schwarzinger, Roberto Mauricio Carvalho Guedes, Paula Angelica Correia, Stefan Hirtenlehner, Christine Potthast, Gerald Klanert, Julian Weghuber
Summary: Lignans as feed supplements have antioxidant, anti-inflammatory, and barrier-strengthening effects on intestinal health.
Article
Chemistry, Multidisciplinary
M. Florencia Sanchez, Marina S. Dietz, Ulrike Mueller, Julian Weghuber, Karl Gatterdam, Ralph Wieneke, Mike Heilemann, Peter Lanzerstorfer, Robert Tampe
Summary: Membrane receptor clustering is important for cell-cell communication, but its physiological role in cell signaling is still unclear. In this study, the researchers developed a dynamic platform to induce cluster formation of neuropeptide Y2 hormone receptors (Y2R) and found that receptor clustering enhanced the cell's response to ligands and played a key role in cell signaling.
Article
Biochemistry & Molecular Biology
Bernhard Blank-Landeshammer, Gerald Klanert, Lisa Mitter, Sophia Turisser, Nicolas Nusser, Alice Koenig, Marcus Iken, Julian Weghuber
Summary: Intestinal absorption of lipophilic micronutrients and phytochemicals is usually low, but plant extracts can enhance their bioavailability. This study investigated the effects of extracts from curcuma, ginger, and black pepper on the uptake and transport rates of lutein, zeaxanthin, and soy isoflavones. It was found that curcuma extract increased lutein uptake, while curcuma and ginger extracts enhanced isoflavone transport rates.
Article
Materials Science, Multidisciplinary
Roland Hager, Christian Forsich, Jiri Duchoslav, Christoph Burgstaller, David Stifter, Julian Weghuber, Peter Lanzerstorfer
Summary: Polymeric materials have emerged as important components in biosensing interfaces, providing superior surfaces for biomolecule binding and printing. This study characterized 11 different polymer foils and evaluated their suitability for surface functionalization, biomolecule micropatterning, and fluorescence microscopy. Results showed that COC, COP, and PMMA were the most promising alternatives to glass coverslips, demonstrating excellent applicability in subcellular micropatterning experiments.
ACS APPLIED POLYMER MATERIALS
(2022)
Article
Chemistry, Multidisciplinary
Saanfor Hubert Suh, Yongzheng Xing, Alexia Rottensteiner, Rong Zhu, Yoo Jin Oh, Stefan Howorka, Peter Hinterdorfer
Summary: This study investigates molecular recognition in a synthetic confined nanopore with controllable pore dimension and molecular DNA receptors at different depth positions. The results show that the association rates are slower and the dissociation rates are faster for receptors positioned deeper inside the pore lumen. This phenomenon can be explained by the steric constraints on molecular interactions in confinement.
Article
Virology
Antonio Real-Hohn, Martin Groznica, Georg Kontaxis, Rong Zhu, Otavio Augusto Chaves, Leonardo Vazquez, Peter Hinterdorfer, Heinrich Kowalski, Dieter Blaas
Summary: Rhinoviruses are the main cause of the common cold and can lead to complications in patients with other ailments. Colds have a significant socioeconomic impact, but there are no vaccines or approved treatments available.
Article
Virology
Markus Kastner, Andreas Karner, Rong Zhu, Qiang Huang, Andreas Geissner, Anne Sadewasser, Markus Lesch, Xenia Woermann, Alexander Karlas, Peter H. Seeberger, Thorsten Wolff, Peter Hinterdorfer, Andreas Herrmann, Christian Sieben
Summary: The study compared influenza virus receptor specificity with virus binding to live cells using glycan arrays and single-virus force spectroscopy. The findings revealed that the receptor binding preference of the virus does not necessarily reflect its specificity to cells. Single-virus force spectroscopy was proposed as a tool to understand the cell binding preference of influenza A viruses.
Article
Food Science & Technology
Nicole Ollinger, Alexandra Malachova, Michael Sulyok, Lisa Schutz-Kapl, Nicole Wiesinger, Rudolf Krska, Julian Weghuber
Summary: In this study, three reliable methods were used to identify mould species in food, aiming to estimate the safety of apparently non-moldy slices of bread and reduce food waste. The results showed that it might be safe to consume bread slices from lower layers of bread stacks, even if mould growth is visible on the outer slices.
