Article
Optics
Zhaoning Zhang, Weibing Kuang, Bing Shi, Zhen - Li Huang
Summary: Colorimetry camera-based fluorescence microscopy (CCFM) is a single-frame imaging method for observing multiple biological events simultaneously. However, under low signal levels, CCFM may have poor color resolution due to high crosstalk, which can be addressed through new algorithms and dye selection methods.
Article
Cell Biology
Clara-Marie Guerth, Maria Augusta do Rego Barros Fernandes Lima, Victor Macarron Palacios, Angel Rafael Cereceda Delgado, Jasmine Hubrich, Elisa D'Este
Summary: This study quantified the occurrence of neurofilament isoforms at the postsynapse and compared them with markers of synaptic strength and activity. The results showed that different isoforms rarely colocalize with each other within dendritic spines, and NFL is the most abundant isoform. The presence of neurofilaments at the postsynapse correlates with synaptic status, shedding new light on their regulation and possible contribution to synaptopathies.
Article
Optics
Angela Pizzuto, Pingchuan Ma, Daniel M. Mittleman
Summary: The coupling of terahertz optical techniques to scattering-type scanning near-field microscopy (s-SNOM) has opened up new possibilities for studying nanoscale phenomena in wide bandgap materials. In this study, researchers successfully demonstrated s-SNOM using blue light, which allowed for the generation of terahertz pulses directly from bulk silicon with nanoscale resolution. This breakthrough enables the extraction of spectroscopic information that is not obtainable with near-infrared excitation, providing a new realm of possibilities for studying wide-bandgap materials.
LIGHT-SCIENCE & APPLICATIONS
(2023)
Article
Biochemical Research Methods
Yuxuan Zhao, Meng Zhang, Wenting Zhang, Yao Zhou, Longbiao Chen, Qing Liu, Peng Wang, Rong Chen, Xinxin Duan, Feifan Chen, Huan Deng, Yunfei Wei, Peng Fei, Yu-Hui Zhang
Summary: Long-term visualization of dynamic interactions between intracellular structures in live cells is critical for understanding cellular functions. This study presents a novel approach that combines deep-learning super-resolution strategy and double-ring-modulated selective plane illumination microscopy design to achieve fast and high-resolution imaging of organelle interactions and dynamics in live cells.
Review
Physiology
Jorik H. Amesz, Lu Zhang, Bian R. Everts, Natasja M. S. De Groot, Yannick J. H. J. Taverne
Summary: Living myocardial slices (LMS) offer a comprehensive model for cardiac arrhythmia research. The introduction of biomimetic electromechanical cultivation chambers has facilitated long-term cultivation and control of living myocardial slices culture conditions. This review aims to present the potential of this biomimetic interface in electrophysiological studies,discussing advantages, disadvantages, and future perspectives of the model, as well as various electrophysiological techniques and their application on living myocardial slices.
FRONTIERS IN PHYSIOLOGY
(2023)
Article
Optics
Jianwei Chen, Benxi Yao, Zhichao Yang, Wei Shi, Tingdan Luo, Peng Xi, Dayong Jin, Yiming Li
Summary: In this study, a new ratiometric multicolor imaging strategy for 4Pi-SMLM was developed, achieving optimal 3D resolution and color separation.
Article
Optics
M. Chen, T. V. A. G. de Oliveira, I. Ilyakov, T. Noerenberg, F. Kuschewski, J-C Deinert, N. Awari, A. Ponomaryov, M. Kuntzsch, S. C. Kehr, L. M. Eng, M. Gensch, S. Kovalev
Summary: This article presents a conceptually new approach to synchronizing accelerator-based light sources and external laser systems. The method utilizes an intense accelerator-based single-cycle terahertz pulse to slice a femtosecond-level part of a longer picosecond laser pulse, achieving precise synchronization of approximately 10 fs for real-time lock-in amplifier signal demodulation. The concept was successfully demonstrated in three benchmark experiments using a 4th generation accelerator-based terahertz light source.
Article
Medicine, General & Internal
Enrico Ripamonti, Arvid Eden, Staffan Nilsson, Anders Sonnerborg, Henrik Zetterberg, Magnus Gisslen
Summary: This retrospective follow-up study investigates the longitudinal change of plasma neurofilament light (NfL) levels after antiretroviral therapy (ART) initiation in people living with HIV. The results show that plasma NfL levels are higher at baseline and decline faster during the follow-up in patients with CD4(+) count 100 cells/μl or higher. There is no significant correlation between baseline HIV RNA levels, treatment regimen, and sex with plasma NfL levels.
JOURNAL OF INTERNAL MEDICINE
(2023)
Article
Chemistry, Multidisciplinary
Kai Xie, Zixun Wang, Lin Qi, Xi Zhao, Yuan Wang, Jin Qu, Ping Xu, Linfeng Huang, Wenjun Zhang, Yang Yang, Xin Wang, Peng Shi
Summary: The described platform allows for multiplexed in situ miRNA profiling in acute tissue slices, providing relative spatial dynamics of cellular miRNAs in associated cell populations. The technique isolates targeted miRNAs from the cytosol of a large population of cells to achieve quasi-single-cell analysis for the tissue sample.
Article
Multidisciplinary Sciences
Tianyan Liu, Till Stephan, Peng Chen, Jan Keller-Findeisen, Jingting Chen, Dietmar Riedel, Zhongtian Yang, Stefan Jakobs, Zhixing Chen
Summary: This study reports a mitochondrial inner membrane fluorescent marker, PK Mito Orange (PKMO), which features reduced phototoxicity and high stability, enabling super-resolution recordings of mitochondrial inner membrane dynamics. The PKMO dye is optically orthogonal with green and far-red markers, allowing multiplexed recordings. By using multi-color STED microscopy, interactions between mitochondria and other cellular components can be captured at sub-100 nm resolution.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2022)
Article
Biotechnology & Applied Microbiology
R. Nunez-Toldra, A. Del Canizo, I. Secco, L. Nicastro, M. Giacca, C. M. Terracciano
Summary: Gene therapy based on viral vectors has great potential for the study and treatment of cardiac diseases. This study explores the use of Living Myocardial Slices (LMS) as a platform for nucleic acid-based therapies, using rat and human cardiac tissues to optimize and analyze gene transfer efficiency.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
Article
Pharmacology & Pharmacy
Qin Wu, Abigail J. Ross, Tugce Ipek, Georgina H. Thompson, Robert D. Johnson, Changhao Wu, Patrizia Camelliti
Summary: The cardiotoxicity risk of hydroxychloroquine (HCQ) and azithromycin (AZM) has been extensively studied due to safety concerns in COVID-19 patients. While these drugs have been associated with QT interval prolongation and drug-induced arrhythmias, other mechanisms of cardiotoxicity remain largely unexplored.
FRONTIERS IN PHARMACOLOGY
(2023)
Article
Endocrinology & Metabolism
Luciana Mateus Goncalves, Joana Almaca
Summary: The study utilized pancreas slices from non-diabetic human donors to investigate the function of human islet blood vessels, revealing the vascular responses to vasoactive stimuli and the critical role of mural cells in regulating blood perfusion in islets. The findings highlight the differences between human and rodent islet microvasculature and emphasize the importance of understanding these alterations in disease progression.
FRONTIERS IN ENDOCRINOLOGY
(2021)
Article
Multidisciplinary Sciences
Xin Guo, Rui Pu, Zhimin Zhu, Shuqian Qiao, Yusen Liang, Bingru Huang, Haichun Liu, Lucia Labrador-Paez, Uliana Kostiv, Pu Zhao, Qiusheng Wu, Jerker Widengren, Qiuqiang Zhan
Summary: The authors introduce a new nanoscopy technique called stimulated-emission induced excitation depletion (STExD), which uses a single pair of low-power, near-infrared, continuous-wave lasers to inhibit the emission of multichromatic probes. By manipulating the common sensitizer in lanthanide upconversion systems, emission inhibition is achieved for a wide range of emitters, with high depletion efficiency and low saturation intensity.
NATURE COMMUNICATIONS
(2022)
Article
Optics
Wenhao Zhang, Kaidi Zhang, Wenting Wang, Yuhang Chen
Summary: Using s-SNOM and energy dispersive spectroscopy, we studied the effects of ion implantation in a silicon substrate, finding correlations between contrast and residual element concentration with ion dose, indicating the sensitivity of s-SNOM imaging to implanted ions.
Article
Biotechnology & Applied Microbiology
Kanuj Mishra, Juan Pablo Fuenzalida-Werner, Francesca Pennacchietti, Robert Janowski, Andriy Chmyrov, Yuanhui Huang, Christian Zakian, Uwe Klemm, Ilaria Testa, Dierk Niessing, Vasilis Ntziachristos, Andre C. Stiel
Summary: Reversibly photo-switchable proteins have been utilized for super-resolution and optoacoustic imaging methods, and in this study, a prototype of a photo-switchable Ca2+ sensor was constructed based on GCaMP5G. The molecular mechanisms of the sensor were described at the structural level, demonstrating the ability to image calcium and other analytes at super-resolution and in vivo.
NATURE BIOTECHNOLOGY
(2022)
Article
Biology
Lea Rems, Xinru Tang, Fangwei Zhao, Sergio Perez-Conesa, Ilaria Testa, Lucie Delemotte
Summary: The study investigates the molecular mechanisms of electroporation on the plasma membrane of biological cells. By combining molecular dynamics simulations, machine learning, and Bayesian survival analysis, the study identifies that the formation of pores depends on the local lipid organization and specific features of membrane domains. The study also reveals the importance of membrane mechanical properties and the polarity of the electric field in pore formation. The findings provide valuable insights for the design of future experiments and the development of accurate descriptions of plasma membrane electroporation.
Article
Biology
Waja Wegner, Heinz Steffens, Carola Gregor, Fred Wolf, Katrin Willig, Yukiko Goda
Summary: This study investigated the effects of environmental enrichment on the morphology and dynamics of individual synapses. The results showed that environmental enrichment can enhance the uniformity of synaptic strength and lead to a more dynamic topography of the PSD95 nanoorganization.
Article
Biochemical Research Methods
Mark Bates, Jan Keller-Findeisen, Adrian Przybylski, Andreas Hueper, Till Stephan, Peter Ilgen, Angel R. Cereceda Delgado, Elisa D'Este, Alexander Egner, Stefan Jakobs, Steffen J. Sahl, Stefan W. Hell
Summary: The dynamic model of the 4Pi point spread function allows for high three-dimensional resolution in localization microscopy. 4Pi-STORM imaging reveals nanoscale details of protein and nucleic acid organization in neurons and mitochondria. Despite challenges in instrumentation and data analysis, the development of a 4Pi-STORM microscope with dynamic PSF modeling and simpler optical design offers optimal resolution and accuracy for molecular organization studies.
Article
Chemistry, Physical
Jonatan Alvelid, Andrea Bucci, Ilaria Testa
Summary: This study introduces a multicolour STED nanoscopy technique using far red-shifted semiconductor CdTe quantum dots (QDs). By optimizing the STED imaging of QDs, blinking effects are minimized and the number of detected photons is maximized. This method enables straightforward three-colour STED imaging and can be applied to study the internalization of QDs in cells.
Article
Biotechnology & Applied Microbiology
Andrea Volpato, Dirk Ollech, Jonatan Alvelid, Martina Damenti, Barbara Mueller, Andrew G. York, Maria Ingaramo, Ilaria Testa
Summary: The authors introduce a method called STARSS, which uses reversible molecular transitions of switchable fluorescent proteins to resolve the rotational diffusion rate of large macromolecular complexes, thus extending the observable mass range. They applied STARSS to investigate the rotational mobility of several molecular complexes in cells.
NATURE BIOTECHNOLOGY
(2023)
Article
Biochemical Research Methods
Lynn M. Ostersehlt, Daniel C. Jans, Anna Wittek, Jan Keller-Findeisen, Kaushik Inamdar, Steffen J. Sahl, Stefan W. Hell, Stefan Jakobs
Summary: The combination of MINFLUX nanoscopy with DNA-PAINT labeling enables multi-color imaging of multiple molecular targets, leading to improved nanoscopy in fixed cells and increased multiplexing in MINFLUX imaging, as demonstrated by three-color imaging of mitochondria in mammalian cells.
Article
Biochemical Research Methods
Jonatan Alvelid, Martina Damenti, Chiara Sgattoni, Ilaria Testa
Summary: Event-triggered STED is an automated imaging method that rapidly initiates STED imaging after detecting cellular events, which can enhance live cell imaging capabilities and enable real-time biological observations and discoveries.
Article
Multidisciplinary Sciences
Elisabeth Grimm, Franciscus van der Hoeven, Donato Sardella, Katrin Willig, Ulrike Engel, Nisha Veits, Robert Engel, Elisabetta Ada Cavalcanti-Adam, Felix Bestvater, Luca Bordoni, Richard Jennemann, Kai Schoenig, Ina Maria Schiessl, Roger Sandhoff
Summary: Clathrin-mediated endocytosis is a well-studied cellular uptake pathway, but the dynamic cooperation of its components in vivo is still not well understood. This study generated a reporter mouse model to track endocytosis by fusing eGFP to clathrin light chain a. The fusion protein was expressed in all tissues and could be visualized using fluorescence microscopy. The functionality of the reporter was confirmed by recruitment to nanobeads recorded by TIRF microscopy.
Article
Multidisciplinary Sciences
Katrin Willig
Summary: Super-resolution fluorescence microscopy has been widely used in biological laboratories and can be applied to imaging the brain of living mice to study sub-cellular structures and brain functions. These studies demonstrate the potential of in vivo super-resolution imaging and provide evidence for investigating the role of nanostructures in memory and learning.
Article
Neurosciences
Martin Fuhrmann, Nala Gockel, Misa Arizono, Yulia Dembitskaya, U. Valentin Naegerl, Francesca Pennacchietti, Martina Damenti, Ilaria Testa, Katrin I. Willig
Summary: Super-resolution fluorescence microscopy has great potential in neuroscience for visualizing nanoscale structures and biochemical activities. Recent technical progress has enhanced the performance and accessibility of this technology, but in vivo applications still face challenges. This review presents vivid examples based on STED and RESOLFT super-resolution microscopy, providing insights into the nanoscale mechanisms of cellular communication between neurons and glia.
JOURNAL OF NEUROSCIENCE
(2022)
Article
Neurosciences
Ajit Ray, Joseph A. Christian, Matthew B. Mosso, Eunsol Park, Waja Wegner, Katrin I. Willig, Alison L. Barth
Summary: High-throughput anatomic data can generate new hypotheses about neural circuit changes. This study used fluorescence-based reagents to examine synapse changes in somatosensory cortex during whisker-dependent learning. The researchers found rapid morphologic changes in thalamocortical synapses at the early stages of training.
JOURNAL OF NEUROSCIENCE
(2023)
Correction
Biology
Vahid Ebrahimi, Till Stephan, Jiah Kim, Pablo Carravilla, Christian Eggeling, Stefan Jakobs, Kyu Young Han
COMMUNICATIONS BIOLOGY
(2023)
Article
Biology
Vahid Ebrahimi, Till Stephan, Jiah Kim, Pablo Carravilla, Christian Eggeling, Stefan Jakobs, Kyu Young Han
Summary: This study demonstrates that restoring STED microscopy images using deep learning can reduce the effects of photobleaching and photodamage by decreasing the pixel dwell time. Our method effectively and reliably restores noisy 2D and 3D STED images, enabling long-term observation of mitochondrial dynamics.
COMMUNICATIONS BIOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Dojin Kim, Stefan Stoldt, Michael Weber, Stefan Jakobs, Vladimir N. N. Belov, Stefan W. W. Hell
Summary: Disulfonated rhodamines, commonly used in life science and optical microscopy, were previously thought to be impermeable to living cells. This assumption was challenged by using five popular rhodamines combined with a HaloTag (TM) amine (O-2) ligand (x) to successfully label living cells. Three compounds with two negative charges, Rho590-x, Rho565-x, and Rho530-x, showed specific and bright staining in living cells and performed well in STED microscopy. Other probes with one negative charge, prepared by native chemical ligation and esterification, exhibited specific staining and red shifts in absorption and emission bands.