The Experts below are selected from a list of 15156 Experts worldwide ranked by ideXlab platform
Johanna Ivaska - One of the best experts on this subject based on the ideXlab platform.
-
Fluctuation-Based Super-Resolution Traction Force Microscopy.
Nano letters, 2020Co-Authors: Aki Stubb, Mitro Miihkinen, Camilo Guzman, Ricardo Henriques, Guillaume Jacquemet, Hellyeh Hamidi, Romain F. Laine, Johanna IvaskaAbstract:Cellular mechanics play a crucial role in tissue homeostasis and are often misregulated in disease. Traction Force microscopy is one of the key methods that has enabled researchers to study fundamental aspects of mechanobiology; however, Traction Force microscopy is limited by poor resolution. Here, we propose a simplified protocol and imaging strategy that enhances the output of Traction Force microscopy by increasing i) achievable bead density and ii) the accuracy of bead tracking. Our approach relies on super-resolution microscopy, enabled by fluorescence fluctuation analysis. Our pipeline can be used on spinning-disk confocal or widefield microscopes and is compatible with available analysis software. In addition, we demonstrate that our workflow can be used to gain biologically relevant information and is suitable for fast long-term live measurement of Traction Forces even in light-sensitive cells. Finally, using fluctuation-based Traction Force microscopy, we observe that filopodia align to the Force field generated by focal adhesions.
-
fluctuation based super resolution Traction Force microscopy
Nano Letters, 2020Co-Authors: Aki Stubb, Mitro Miihkinen, Camilo Guzman, Ricardo Henriques, Guillaume Jacquemet, Hellyeh Hamidi, Romain F. Laine, Johanna IvaskaAbstract:Cellular mechanics play a crucial role in tissue homeostasis and are often misregulated in disease. Traction Force microscopy is one of the key methods that has enabled researchers to study fundame...
Huw Colinyork - One of the best experts on this subject based on the ideXlab platform.
-
spatiotemporally super resolved volumetric Traction Force microscopy
Nano Letters, 2019Co-Authors: Huw Colinyork, Yousef Javanmardi, Liliana Barbieri, Kseniya Korobchevskaya, Chloe M Hall, Aaron B Taylor, Satya Khuon, Graham K Sheridan, Di Li, Tengleong ChewAbstract:Quantification of mechanical Forces is a major challenge across biomedical sciences. Yet such measurements are essential to understanding the role of biomechanics in cell regulation and function. Traction Force microscopy remains the most broadly applied Force probing technology but typically restricts itself to single-plane two-dimensional quantifications with limited spatiotemporal resolution. Here, we introduce an enhanced Force measurement technique combining 3D super-resolution fluorescence structural illumination microscopy and Traction Force microscopy (3D-SIM-TFM) offering increased spatiotemporal resolution, opening-up unprecedented insights into physiological three-dimensional Force production in living cells.
-
the future of Traction Force microscopy
Current Opinion in Biomedical Engineering, 2018Co-Authors: Huw Colinyork, Marco FritzscheAbstract:Abstract Animal cells continuously sense and respond to mechanical Force. Quantifying these Forces remains a major challenge in bioengineering; yet such measurements are essential for the understanding of cellular function. Traction Force microscopy is one of the most successful and broadly-used Force probing technologies, chosen for the simplicity of its implementation, flexibility to mimic cellular conditions, and well-established analysis pipe-line. Here, we review the accomplishments, and discuss the applicability and limitations of Traction Force microscopy. We explain fundamental shortcomings of the method, summarise latest improvements, and outline future pathways towards the impact of the method, especially considering latest developments in state-of-the-art super-resolution fluorescence imaging. In light of the increasing discovery of the importance of mechanobiology in cell physiology, we envisage Traction Force microscopy to remain a major player for quantifying mechanical Forces in living cells.
-
super resolved Traction Force microscopy stfm
Nano Letters, 2016Co-Authors: Huw Colinyork, Dilip Shrestha, James H Felce, Dominic Waithe, Emad Moeendarbary, Simon J Davis, Christian Eggeling, Marco FritzscheAbstract:Measuring small Forces is a major challenge in cell biology. Here we improve the spatial resolution and accuracy of Force reconstruction of the well-established technique of Traction Force microscopy (TFM) using STED microscopy. The increased spatial resolution of STED-TFM (STFM) allows a greater than 5-fold higher sampling of the Forces generated by the cell than conventional TFM, accessing the nano instead of the micron scale. This improvement is highlighted by computer simulations and an activating RBL cell model system.
Cai Huang - One of the best experts on this subject based on the ideXlab platform.
-
Roles of Talin2 in Traction Force Generation, Tumor Metastasis and Cardiovascular Integrity.
Current protein & peptide science, 2018Co-Authors: Tomasz Kolodziej, Zenon Rajfur, Cai HuangAbstract:There are two vertebrate talin genes, TLN1 and TLN2, which encode talin1 and talin2. Talin1 governs integrin activation, thus regulating focal adhesion (FA) assembly, cell migration and invasion, but the biological function of talin2 remains to be elucidated and not too long ago talin2 was presumed to function redundantly with talin1. Recent studies have shown distinct differences between talin2 and talin1. The promoter of TLN2 is different from that of TLN1 in their size and binding to different transcription factors. Talin2 has a higher affinity to β -integrins than talin1. Talin2 regulates Traction Force generation, focal adhesion dynamics and invadopodium formation, thus controlling tumor cell migration, invasion and metastasis. Also, talin2 is enriched in the myotendinous junction (MTJ) in striated muscle, costameres and intercalated disks (ICDs) of cardiac myofibrils, and atherosclerotic plaques of blood vessels, thus regulating cardiovascular integrity. In this review, we discuss the differences between talin1 and talin2, in genome, protein expression pattern, affinity with integrins, Traction Force generation, and provide a glance at the roles of talin2 in cancer cell invasion and cardiovascular function.
-
Talin2-mediated Traction Force drives matrix degradation and cell invasion
Journal of cell science, 2016Co-Authors: Naser Jafari, Zaozao Chen, Vesa P. Hytönen, Benjamin T. Goult, Chang-guo Zhan, Cai HuangAbstract:ABSTRACT Talin binds to β-integrin tails to activate integrins, regulating cell migration, invasion and metastasis. There are two talin genes, TLN1 and TLN2, encoding talin1 and talin2, respectively. Talin1 regulates focal adhesion dynamics, cell migration and invasion, whereas the biological function of talin2 is not clear and, indeed, talin2 has been presumed to function redundantly with talin1. Here, we show that talin2 has a much stronger binding to β-integrin tails than talin1. Replacement of talin2 Ser339 with Cys significantly decreased its binding to β1-integrin tails to a level comparable to that of talin1. Talin2 localizes at invadopodia and is indispensable for the generation of Traction Force and invadopodium-mediated matrix degradation. Ablation of talin2 suppressed Traction Force generation and invadopodia formation, which were restored by re-expressing talin2 but not talin1. Furthermore, re-expression of wild-type talin2 (but not talin2S339C) in talin2-depleted cells rescued development of Traction Force and invadopodia. These results suggest that a strong interaction of talin2 with integrins is required to generate Traction, which in turn drives invadopodium-mediated matrix degradation, which is key to cancer cell invasion.
Gwang Lee - One of the best experts on this subject based on the ideXlab platform.
-
Decrease in membrane fluidity and Traction Force induced by silica-coated magnetic nanoparticles.
Journal of nanobiotechnology, 2021Co-Authors: Tae Hwan Shin, Abdurazak Aman Ketebo, Da Yeon Lee, Seungah Lee, Seong Ho Kang, Shaherin Basith, Balachandran Manavalan, Hyeon Kwon, Sungsu Park, Gwang LeeAbstract:BACKGROUND Nanoparticles are being increasingly used in biomedical applications owing to their unique physical and chemical properties and small size. However, their biophysical assessment and evaluation of side-effects remain challenging. We addressed this issue by investigating the effects of silica-coated magnetic nanoparticles containing rhodamine B isothiocyanate [MNPs@SiO2(RITC)] on biophysical aspects, such as membrane fluidity and Traction Force of human embryonic kidney 293 (HEK293) cells. We further extended our understanding on the biophysical effects of nanoparticles on cells using a combination of metabolic profiling and transcriptomic network analysis. RESULTS Overdose (1.0 μg/µL) treatment with MNPs@SiO2(RITC) induced lipid peroxidation and decreased membrane fluidity in HEK293 cells. In addition, HEK293 cells were morphologically shrunk, and their aspect ratio was significantly decreased. We found that each Traction Force (measured in micropillar) was increased, thereby increasing the total Traction Force in MNPs@SiO2(RITC)-treated HEK293 cells. Due to the reduction in membrane fluidity and elevation of Traction Force, the velocity of cell movement was also significantly decreased. Moreover, intracellular level of adenosine triphosphate (ATP) was also decreased in a dose-dependent manner upon treatment with MNPs@SiO2(RITC). To understand these biophysical changes in cells, we analysed the transcriptome and metabolic profiles and generated a metabotranscriptomics network, which revealed relationships among peroxidation of lipids, focal adhesion, cell movement, and related genes and metabolites. Furthermore, in silico prediction of the network showed increment in the peroxidation of lipids and suppression of focal adhesion and cell movement. CONCLUSION Taken together, our results demonstrated that overdose of MNPs@SiO2(RITC) impairs cellular movement, followed by changes in the biophysical properties of cells, thus highlighting the need for biophysical assessment of nanoparticle-induced side-effects.
-
Decrease in Membrane Fluidity and Traction Force Induced by Silica-Coated Magnetic Nanoparticles
2020Co-Authors: Tae Hwan Shin, Abdurazak Aman Ketebo, Seungah Lee, Seong Ho Kang, Shaherin Basith, Balachandran Manavalan, Sungsu Park, Da Yeon Lee, Gwang LeeAbstract:Abstract Background Nanoparticles are being used increasingly due to their unique physical and chemical properties and small size. It is well-known that nanoparticles cause side effects, however their biophysical assessment remains challenging. We addressed this issue by investigating the effects of silica-coated magnetic nanoparticles containing rhodamine B isothiocyanate [MNPs@SiO2(RITC)] on the biophysical aspects, such as membrane fluidity and Traction Force of human embryonic kidney 293 (HEK293) cells. We further extended our understanding on the biophysical effects of nanoparticles on cells using a combination of metabolic profiling and transcriptomic network analysis. Results Overdose (1.0 μg/µl) treatment of MNPs@SiO2(RITC) induced lipid peroxidation and decreased membrane fluidity in HEK293 cells. During membrane damage, HEK293 cells were morphologically shrunk and aspect ratio of the cells were significantly decreased upon MNPs@SiO2(RITC) treatment. Each of Traction Force (measured in micropillar) was found to be increased, thereby increasing the total Traction Force in MNPs@SiO2(RITC)-treated HEK293 cells. Due to the reduction in membrane fluidity and elevation of Traction Force, velocity of the cell movement was significantly decreased in MNPs@SiO2(RITC)-treated HEK293 cells. Moreover, intracellular ATP also decreased in a dose dependent manner upon MNPs@SiO2(RITC) treatment. To understand the biophysical changes in cells, we analysed transcriptome and metabolic profiles and generated metabotranscriptomics network. The network showed relationships among peroxidation of lipid, focal adhesion, cell movement, and related genes and metabolites. Furthermore, in silico prediction of the network showed increment in the peroxidation of lipid and suppression of focal adhesion and cell movement.Conclusion Taken together, our results demonstrate that overdosage of MNPs@SiO2(RITC) impairs cellular movement, followed by changes in the biophysical properties of cells, thus highlighting the need for biophysical assessment of nanoparticle-induced side effects.
Marco Fritzsche - One of the best experts on this subject based on the ideXlab platform.
-
the future of Traction Force microscopy
Current Opinion in Biomedical Engineering, 2018Co-Authors: Huw Colinyork, Marco FritzscheAbstract:Abstract Animal cells continuously sense and respond to mechanical Force. Quantifying these Forces remains a major challenge in bioengineering; yet such measurements are essential for the understanding of cellular function. Traction Force microscopy is one of the most successful and broadly-used Force probing technologies, chosen for the simplicity of its implementation, flexibility to mimic cellular conditions, and well-established analysis pipe-line. Here, we review the accomplishments, and discuss the applicability and limitations of Traction Force microscopy. We explain fundamental shortcomings of the method, summarise latest improvements, and outline future pathways towards the impact of the method, especially considering latest developments in state-of-the-art super-resolution fluorescence imaging. In light of the increasing discovery of the importance of mechanobiology in cell physiology, we envisage Traction Force microscopy to remain a major player for quantifying mechanical Forces in living cells.
-
super resolved Traction Force microscopy stfm
Nano Letters, 2016Co-Authors: Huw Colinyork, Dilip Shrestha, James H Felce, Dominic Waithe, Emad Moeendarbary, Simon J Davis, Christian Eggeling, Marco FritzscheAbstract:Measuring small Forces is a major challenge in cell biology. Here we improve the spatial resolution and accuracy of Force reconstruction of the well-established technique of Traction Force microscopy (TFM) using STED microscopy. The increased spatial resolution of STED-TFM (STFM) allows a greater than 5-fold higher sampling of the Forces generated by the cell than conventional TFM, accessing the nano instead of the micron scale. This improvement is highlighted by computer simulations and an activating RBL cell model system.