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关键词:'tweezers'
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Tinka V M Clement et al.
Methods in molecular biology (Clifton, N.J.), 2694, 91-107 (2023-10-12)
During mitosis, cells compact their DNA into rodlike shapes, four orders of magnitude shorter than the DNA backbone contour length. We describe an experimental protocol to isolate and study these intricate mitotic chromosomes using optical tweezers. We touch upon the
Anna-Karin Gustavsson et al.
Current protocols in cell biology, 82(1), e70-e70 (2018-10-18)
In this unit, we provide a clear exposition of the methodology employed to study dynamic responses in individual cells, using microfluidics for controlling and adjusting the cell environment, optical tweezers for precise cell positioning, and fluorescence microscopy for detecting intracellular
L Gardini et al.
Methods in molecular biology (Clifton, N.J.), 1805, 151-184 (2018-07-05)
Mechanical transitions in molecular motors often occur on a submillisecond time scale and rapidly follow binding of the motor with its cytoskeletal filament. Interactions of nonprocessive molecular motors with their filament can be brief and last for few milliseconds or
Alexey N Semenov et al.
Biomolecules, 10(10) (2020-10-21)
An elevated concentration of fibrinogen in blood is a significant risk factor during many pathological diseases, as it leads to an increase in red blood cells (RBC) aggregation, resulting in hemorheological disorders. Despite the biomedical importance, the mechanisms of fibrinogen-induced
Eleanor M Fletcher et al.
Current protocols, 3(8), e854-e854 (2023-08-09)
Plant organelles are associated with each other through tethering proteins at membrane contact sites (MCS). Methods such as total internal reflection fluorescence (TIRF) optical tweezers allow us to probe organelle interactions in live plant cells. Optical tweezers (focused infrared laser
Artem K Efremov et al.
Biomaterials, 284, 121477-121477 (2022-04-09)
Filopodia are ubiquitous membrane projections that play crucial role in guiding cell migration on rigid substrates and through extracellular matrix by utilizing yet unknown mechanosensing molecular pathways. As recent studies show that Ca2+ channels localized to filopodia play an important
Anna Wypijewska Del Nogal et al.
International journal of molecular sciences, 22(16) (2021-08-28)
Pre-miRNA-377 is a hairpin-shaped regulatory RNA associated with heart failure. Here, we use single-molecule optical tweezers to unzip pre-miRNA-377 and study its stability and dynamics. We show that magnesium ions have a strong stabilizing effect, and that sodium ions stabilize
Rajeev Yadav et al.
Methods in molecular biology (Clifton, N.J.), 2478, 141-240 (2022-09-06)
We present an instrument that combines high-resolution optical tweezers and multicolor confocal fluorescence spectroscopy. Biological macromolecules exhibit complex conformation and stoichiometry changes in coordination with their motion and activity. To further our understanding of the complex machinery of life, we
Kalpana Mandal et al.
Proceedings of the National Academy of Sciences of the United States of America, 113(46), E7159-E7168 (2016-11-02)
The mechanical properties of cells impact on their architecture, their migration, intracellular trafficking, and many other cellular functions and have been shown to be modified during cancer progression. We have developed an approach to map the intracellular mechanical properties of
Varsha P Desai et al.
Molecular cell, 75(5), 1007-1019 (2019-09-01)
The movement of ribosomes on mRNA is often interrupted by secondary structures that present mechanical barriers and play a central role in translation regulation. We investigate how ribosomes couple their internal conformational changes with the activity of translocation factor EF-G
Yuxin Hao et al.
Methods in molecular biology (Clifton, N.J.), 2394, 485-498 (2022-01-31)
Mutations in protein kinases are often associated with the development of cancer, and application of mutant-specific inhibitors as therapeutic measures have shown a remarkable improvement in prolonging patient survival. However, it has also been observed that tumors bearing certain mutation
Tung T Le et al.
Cell, 179(3), 619-631 (2019-10-19)
DNA replication in eukaryotes generates DNA supercoiling, which may intertwine (braid) daughter chromatin fibers to form precatenanes, posing topological challenges during chromosome segregation. The mechanisms that limit precatenane formation remain unclear. By making direct torque measurements, we demonstrate that the
Toyotaka Ishibashi et al.
Proceedings of the National Academy of Sciences of the United States of America, 111(9), 3419-3424 (2014-02-20)
Transcription factors IIS (TFIIS) and IIF (TFIIF) are known to stimulate transcription elongation. Here, we use a single-molecule transcription elongation assay to study the effects of both factors. We find that these transcription factors enhance overall transcription elongation by reducing
Hannah M Stephens et al.
Methods in molecular biology (Clifton, N.J.), 2478, 727-753 (2022-09-06)
T-cell antigen receptors (TCRs) are mechanosensors, which initiate a signaling cascade upon ligand recognition resulting in T-cell differentiation, homeostasis, effector and regulatory functions. An optical trap combined with fluorescence permits direct monitoring of T-cell triggering in response to force application
Eva Vergucht et al.
Scientific reports, 5, 9049-9049 (2015-03-13)
We report on a radically new elemental imaging approach for the analysis of biological model organisms and single cells in their natural, in vivo state. The methodology combines optical tweezers (OT) technology for non-contact, laser-based sample manipulation with synchrotron radiation
Juan Carlos Cordova et al.
Methods in molecular biology (Clifton, N.J.), 1486, 317-341 (2016-11-16)
Energy-dependent protein degradation is studied through the dual bead ClpXP motility assay. Processing of folded proteins involves recognition, unfolding, translocation, and degradation stages. A dual optical trap, in a passive force-clamp geometry, exhibits bead-to-bead displacements that directly follow subprocesses underlying
Jerome Pine et al.
IEEE transactions on bio-medical engineering, 56(4), 1184-1188 (2009-03-11)
The use of an optical tweezer for moving dissociated neurons was studied. The main features of the tweezers are outlined as well as the general principles of its operation. Infrared beams at 980 and 1064 nm were used, focused so
Matthew P Nicholas et al.
Methods in molecular biology (Clifton, N.J.), 1136, 171-246 (2014-03-19)
Numerous microtubule-associated molecular motors, including several kinesins and cytoplasmic dynein, produce opposing forces that regulate spindle and chromosome positioning during mitosis. The motility and force generation of these motors are therefore critical to normal cell division, and dysfunction of these
Songyu Hu et al.
Journal of biomedical nanotechnology, 9(2), 281-285 (2013-05-01)
This paper presents an approach to manipulating cell adhesions using optical tweezers for cell-to-cell interactions at single cell level. A case study of investigating the adhesions between leukemia cells and bone marrow stromal cells is reported. First, the trapping force
Michael J Greenberg et al.
Methods in molecular biology (Clifton, N.J.), 1486, 483-509 (2016-11-16)
The myosin superfamily of molecular motors utilizes energy from ATP hydrolysis to generate force and motility along actin filaments in a diverse array of cellular processes. These motors are structurally, kinetically, and mechanically tuned to their specific molecular roles in
Eric M Patrick et al.
Nature chemical biology, 16(7), 801-809 (2020-02-19)
Telomere maintenance by telomerase is essential for continuous proliferation of human cells and is vital for the survival of stem cells and 90% of cancer cells. To compensate for telomeric DNA lost during DNA replication, telomerase processively adds GGTTAG repeats
Nga Tsing Tang et al.
The Analyst, 148(17), 4099-4108 (2023-07-24)
Bladder cancer is a common cancer that is relatively hard to detect at an early stage because of its non-obvious symptoms. It is known that bladder cells can be found in urine samples which potentially could be used for early
Lucia Gardini et al.
Methods in molecular biology (Clifton, N.J.), 2478, 513-557 (2022-09-06)
Interactions between biological molecules occur on very different time scales, from the minutes of strong protein-protein bonds, down to below the millisecond duration of rapid biomolecular interactions. Conformational changes occurring on sub-ms time scales and their mechanical force dependence underlie
Lu Rao et al.
Methods in molecular biology (Clifton, N.J.), 1665, 53-89 (2017-09-25)
Cytoplasmic dynein is the largest and most intricate cytoskeletal motor protein. It is responsible for a vast array of biological functions, ranging from the transport of organelles and mRNAs to the movement of nuclei during neuronal migration and the formation
Špela Zemljič Jokhadar et al.
PloS one, 15(12), e0231606-e0231606 (2021-01-01)
Keratin intermediate filaments are the principal structural element of epithelial cells. Their importance in providing bulk cellular stiffness is well recognized, but their role in the mechanics of cell cortex is less understood. In this study, we therefore compared the
Julian Kirch et al.
Proceedings of the National Academy of Sciences of the United States of America, 109(45), 18355-18360 (2012-10-24)
In this study, the mobility of nanoparticles in mucus and similar hydrogels as model systems was assessed to elucidate the link between microscopic diffusion behavior and macroscopic penetration of such gels. Differences in particle adhesion to mucus components were strongly
Pooja Patheja et al.
Journal of biophotonics, 8(9), 694-704 (2014-10-31)
The use of optical trap and microbeam for investigating mechanical and transport properties of inter cellular tunneling nanotubes (TnTs) in tumor spheroids has been demonstrated. TnTs in tumor spheroids have been visualized by manipulating TnT connected cells using optical tweezers.
Kalpana Mandal et al.
Nano letters, 19(11), 7691-7702 (2019-10-01)
Molecular motors play important roles in force generation, migration, and intracellular trafficking. Changes in specific motor activities are altered in numerous diseases. KIF20A, a motor protein of the kinesin-6 family, is overexpressed in bladder cancer, and KIF20A levels correlate negatively
Richard W Bowman et al.
Physical review letters, 110(9), 095902-095902 (2013-03-19)
Diamond anvil cells allow the behavior of materials to be studied at pressures up to hundreds of gigapascals in a small and convenient instrument. However, physical access to the sample is impossible once it is pressurized. We show that optical
Heloise P Fernandes et al.
Scandinavian journal of clinical and laboratory investigation, 73(3), 262-264 (2013-02-14)
Classic immunohematology approaches, based on agglutination techniques, have been used in manual and automated immunohematology laboratory routines. Red blood cell (RBC) agglutination depends on intermolecular attractive forces (hydrophobic bonds, Van der Walls, electrostatic forces and hydrogen bonds) and repulsive interactions
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