Optical manipulation of single molecules in the living cell

2014 ◽  
Vol 16 (25) ◽  
pp. 12614-12624 ◽  
Author(s):  
Kamilla Norregaard ◽  
Liselotte Jauffred ◽  
Kirstine Berg-Sørensen ◽  
Lene B. Oddershede
Keyword(s):  
Optical Tweezers ◽  
Living Cell ◽  
Single Molecules ◽  
Optical Manipulation ◽  
Force Measurements

Optical tweezers are the only nano-tools capable of manipulating and performing force-measurements on individual molecules and organelles inside the living cell. We present methodologies for in vivo calibration and exciting recent results.

scholarly journals Hemodynamic forces can be accurately measured in vivo with optical tweezers

2017 ◽  
Author(s):  
Sébastien Harlepp ◽  
Fabrice Thalmann ◽  
Gautier Follain ◽  
Jacky G. Goetz
Keyword(s):  
Optical Tweezers ◽  
Cell Biology ◽  
Accurate Determination ◽  
Force Measurements ◽  
Hemodynamic Forces ◽  
Non Invasive ◽  
Drag Forces ◽  
Measured Flow ◽  
Living Organisms

AbstractForce sensing and generation at the tissular and cellular scale is central to many biological events. There is a growing interest in modern cell biology for methods enabling force measurements in vivo. Optical trapping allows non-invasive probing of pico-Newton forces and thus emerged as a promising mean for assessing biomechanics in vivo. Nevertheless, the main obstacles rely in the accurate determination of the trap stiffness in heterogeneous living organisms, at any position where the trap is used. A proper calibration of the trap stiffness is thus required for performing accurate and reliable force measurements in vivo. Here, we introduce a method that overcomes these difficulties by accurately measuring hemodynamic profiles in order to calibrate the trap stiffness. Doing so, and using numerical methods to assess the accuracy of the experimental data, we measured flow profiles and drag forces imposed to trapped red blood cells of living zebrafish embryos. Using treatments enabling blood flow tuning, we demonstrated that such method is powerful in measuring hemodynamic forces in vivo with accuracy and confidence. Altogether, this study demonstrates the power of optical tweezing in measuring low range hemodynamic forces in vivo and offers an unprecedented tool in both cell and developmental biology.

scholarly journals Calibration of Optical Tweezers for In Vivo Force Measurements: How do Different Approaches Compare?

2014 ◽  
Vol 107 (6) ◽  
pp. 1474-1484 ◽  
Author(s):  
Yonggun Jun ◽  
Suvranta K. Tripathy ◽  
Babu R.J. Narayanareddy ◽  
Michelle K. Mattson-Hoss ◽  
Steven P. Gross
Keyword(s):  
Optical Tweezers ◽  
Force Measurements

scholarly journals Hemodynamic forces can be accurately measured in vivo with optical tweezers

2017 ◽  
Vol 28 (23) ◽  
pp. 3252-3260 ◽  
Author(s):  
Sébastien Harlepp ◽  
Fabrice Thalmann ◽  
Gautier Follain ◽  
Jacky G. Goetz
Keyword(s):  
Optical Tweezers ◽  
Cell Biology ◽  
Accurate Determination ◽  
Force Measurements ◽  
Hemodynamic Forces ◽  
Drag Forces ◽  
Measured Flow ◽  
Living Organisms

Force sensing and generation at the tissue and cellular scale is central to many biological events. There is a growing interest in modern cell biology for methods enabling force measurements in vivo. Optical trapping allows noninvasive probing of piconewton forces and thus emerged as a promising mean for assessing biomechanics in vivo. Nevertheless, the main obstacles lie in the accurate determination of the trap stiffness in heterogeneous living organisms, at any position where the trap is used. A proper calibration of the trap stiffness is thus required for performing accurate and reliable force measurements in vivo. Here we introduce a method that overcomes these difficulties by accurately measuring hemodynamic profiles in order to calibrate the trap stiffness. Doing so, and using numerical methods to assess the accuracy of the experimental data, we measured flow profiles and drag forces imposed to trapped red blood cells of living zebrafish embryos. Using treatments enabling blood flow tuning, we demonstrated that such a method is powerful in measuring hemodynamic forces in vivo with accuracy and confidence. Altogether this study demonstrates the power of optical tweezing in measuring low range hemodynamic forces in vivo and offers an unprecedented tool in both cell and developmental biology.

Solving the mechanisms responsible for organelle behavior in vivo by same-cell correlative light and electron microscopy

1992 ◽  
Vol 50 (1) ◽  
pp. 14-15
Author(s):  
Conly L. Rieder
Keyword(s):  
Electron Microscopy ◽  
Quantitative Analysis ◽  
Light Microscopy ◽  
Living Cell ◽  
Light And Electron Microscopy ◽  
Time Behavior ◽  
Dynamic Interactions ◽  
Positive Identification ◽  
Cellular Components

The behavior of many cellular components, and their dynamic interactions, can be characterized in the living cell with considerable spatial and temporal resolution by video-enhanced light microscopy (video-LM). Indeed, under the appropriate conditions video-LM can be used to determine the real-time behavior of organelles ≤ 25-nm in diameter (e.g., individual microtubules—see). However, when pushed to its limit the structures and components observed within the cell by video-LM cannot be resolved nor necessarily even identified, only detected. Positive identification and a quantitative analysis often requires the corresponding electron microcopy (EM).

scholarly journals Nano‐Optical Tweezers: Methods and Applications for Trapping Single Molecules and Nanoparticles

ChemPhysChem ◽  
2021 ◽  
Vol 22 (14) ◽  
pp. 1408-1408
Author(s):  
Joshua D. Kolbow ◽  
Nathan C. Lindquist ◽  
Christopher T. Ertsgaard ◽  
Daehan Yoo ◽  
Sang‐Hyun Oh
Keyword(s):  
Optical Tweezers ◽  
Single Molecules

scholarly journals Force generation by protein–DNA co-condensation

2021 ◽  
Author(s):  
Thomas Quail ◽  
Stefan Golfier ◽  
Maria Elsner ◽  
Keisuke Ishihara ◽  
Vasanthanarayan Murugesan ◽  
...  
Keyword(s):  
Optical Tweezers ◽  
Self Assembly ◽  
Spider Silk ◽  
Regulatory Elements ◽  
Heterochromatin Formation ◽  
First Order Phase Transition ◽  
Dna Strand ◽  
First Order Phase

AbstractInteractions between liquids and surfaces generate forces1,2 that are crucial for many processes in biology, physics and engineering, including the motion of insects on the surface of water3, modulation of the material properties of spider silk4 and self-assembly of microstructures5. Recent studies have shown that cells assemble biomolecular condensates via phase separation6. In the nucleus, these condensates are thought to drive transcription7, heterochromatin formation8, nucleolus assembly9 and DNA repair10. Here we show that the interaction between liquid-like condensates and DNA generates forces that might play a role in bringing distant regulatory elements of DNA together, a key step in transcriptional regulation. We combine quantitative microscopy, in vitro reconstitution, optical tweezers and theory to show that the transcription factor FoxA1 mediates the condensation of a protein–DNA phase via a mesoscopic first-order phase transition. After nucleation, co-condensation forces drive growth of this phase by pulling non-condensed DNA. Altering the tension on the DNA strand enlarges or dissolves the condensates, revealing their mechanosensitive nature. These findings show that DNA condensation mediated by transcription factors could bring distant regions of DNA into close proximity, suggesting that this physical mechanism is a possible general regulatory principle for chromatin organization that may be relevant in vivo.

Accuracy of Orthodontic Force and Tooth Movement Measurements

1996 ◽  
Vol 23 (3) ◽  
pp. 241-248 ◽  
Author(s):  
Dan Lundgren ◽  
Py Owman-Moll ◽  
Jüri Kurol ◽  
Birgit Mårtensson
Keyword(s):  
Tooth Movement ◽  
Mean Value ◽  
Measurement Methods ◽  
Calibration Data ◽  
Force Measurements ◽  
Accuracy Of Measurement ◽  
Measuring Machine ◽  
Initial Force ◽  
Level Of Agreement

This study was designed to test the accuracy of measurement methods for assessment of force and tooth movement in orthodontic procedures. Daily in vivo measurements of the force produced by activated archwires showed that the initial force declined substantially (by 20 per cent of mean value) within 3 days. Both the ‘trueness’ (validity) and precision of the force measurements, obtained with a strain gauge, were found to be high (SD values were 1·0 cN and 0·4 cN, respectively). Horizontal tooth movements were measured with three different instruments: a slide calliper, a co-ordinate measuring machine, and laser measuring equipment based on holograms. There was a good level of agreement between these methods. This was also confirmed by calibration data. The precision of the methods was (SD values) 0·06, 0·07, and 0·13 mm, respectively. The benefits of the use of the co-ordinate measuring machine are obvious, since it can measure tooth movements in relation to reference planes in all directions.

scholarly journals Rational design of protein-specific folding modifiers

2020 ◽  
Author(s):  
Anirban Das ◽  
Anju Yadav ◽  
Mona Gupta ◽  
R Purushotham ◽  
Vishram L. Terse ◽  
...  
Keyword(s):  
Single Molecule ◽  
Rational Design ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Force Measurements ◽  
Folding Nucleus ◽  
Dynamics Simulations ◽  
General Method

AbstractProtein folding can go wrong in vivo and in vitro, with significant consequences for the living cell and the pharmaceutical industry, respectively. Here we propose a general design principle for constructing small peptide-based protein-specific folding modifiers. We construct a ‘xenonucleus’, which is a pre-folded peptide that resembles the folding nucleus of a protein, and demonstrate its activity on the folding of ubiquitin. Using stopped-flow kinetics, NMR spectroscopy, Förster Resonance Energy transfer, single-molecule force measurements, and molecular dynamics simulations, we show that the ubiquitin xenonucleus can act as an effective decoy for the native folding nucleus. It can make the refolding faster by 33 ± 5% at 3 M GdnHCl. In principle, our approach provides a general method for constructing specific, genetically encodable, folding modifiers for any protein which has a well-defined contiguous folding nucleus.

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