Tuning the Music: Acoustic Force Spectroscopy (AFS) 2.0

Kamsma, Douwe; Creyghton, Ramon; Sitters, Gerrit; Wuite, Gijs J. L.; Peterman, Erwin J. G.

doi:10.1016/j.ymeth.2016.05.002

Cited by 56 publications

(51 citation statements)

References 28 publications

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“…Buoyancy and gravity acting on the cells can be calculated independently on the basis of their size and weight. The acoustic force is in balance with these three forces and can thus be determined for each individual cell (Kamsma et al, 2016) ( Figure S4). Cell velocities are measured during application of successive acoustic pulses of increasing amplitude, spaced by $1 min to allow the cells to sediment back to the bottom of the AFS chamber ( Figure 5A).…”

Section: Calibration Of the Acoustic Force Acting On The Cellsmentioning

confidence: 99%

See 1 more Smart Citation

Single-Cell Acoustic Force Spectroscopy: Resolving Kinetics and Strength of T Cell Adhesion to Fibronectin

Kamsma

Bochet

Oswald³

et al. 2018

Cell Reports

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Assessing the strength and kinetics of molecular interactions of cells with the extracellular matrix is fundamental to understand cell adhesion processes. Given the relevance of these processes, there is a strong need for physical methods to quantitatively assess the mechanism of cell adhesion at the single-cell level, allowing discrimination of cells with different behaviors. Here we introduce single-cell acoustic force spectroscopy (scAFS), an approach that makes use of acoustic waves to exert controlled forces, up to 1 nN, to hundreds of individual cells in parallel. We demonstrate the potential of scAFS by measuring adhesion forces and kinetics of CD4 T lymphocytes (CD4) to fibronectin. We determined that CD4 adhesion is accelerated by interleukin-7, their main regulatory cytokine, whereas CD4 binding strength remains the same. Activation of these cells likely increases their chance to bind to the vessel wall in the blood flow to infiltrate inflamed tissues and locally coordinate the immune response.

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Section: Calibration Of the Acoustic Force Acting On The Cellsmentioning

confidence: 99%

“…The acoustic radiation force F rad was calibrated using the method previously described by Kamsma et al (2016). Here the acoustic force was determined by writing down the force balance during the uplift of unbound cells.…”

Section: Acoustic Force Calibration On Cellsmentioning

confidence: 99%

Single-Cell Acoustic Force Spectroscopy: Resolving Kinetics and Strength of T Cell Adhesion to Fibronectin

Kamsma

Bochet

Oswald³

et al. 2018

Cell Reports

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“…By manipulation of microbeads, single-molecule force spectroscopy techniques revealed the mechanical properties of biomolecules such as DNA or RNA with unprecedented detail [2][3][4][5]. In addition, the interactions with proteins, like the DNA compaction by histones in eukaryotic chromatin [6][7][8][9][10][11] and prokaryotic architectural proteins [12][13][14][15][16], supercoiling [17][18][19][20], and repair processes [21][22][23] were extensively studied with magnetic tweezers (MT) or optical tweezers (OT), Acoustic Force Spectroscopy (AFS) [24][25][26] or tethered particle motion (TPM) [13,27,28]. These bead manipulation techniques have also been used to quantify the mechanical properties of other biological structures, such as extracellular protein collagen [29][30][31], or even entire cells [32].…”

Section: Introductionmentioning

confidence: 99%

Multiplexed Nanometric 3D Tracking of Microbeads using a FFT-Phasor Algorithm

Brouwer

Hermans

Noort

2019

Preprint

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Many single-molecule biophysical techniques rely on nanometric tracking of microbeads to obtain quantitative information about the mechanical properties of biomolecules such as chromatin fibers. Their three-dimensional position can be resolved by holographic analysis of the diffraction pattern in wide-field imaging. Fitting this diffraction pattern to Lorentz Mie scattering theory yields the bead position with nanometer accuracy in three dimensions but is computationally expensive. Real-time multiplexed bead tracking therefore requires a more efficient tracking method. Here, we introduce 3D phasor tracking, a fast and robust bead tracking algorithm with nanometric localization accuracy in arange of over 10 µm. The algorithm is based on a 2D cross-correlation using Fast Fourier Transforms with computer-generated reference images, yielding a processing rate of up to 10.000 regions of interest per second. We implemented the technique in magnetic tweezers and tracked the 3D position of over 100 beads in real-time on a generic CPU. Its easy implementation, efficiency, and robustness can improve multiplexed biophysical bead tracking applications, especially where high throughput is required. SignificanceMicrobeads are often used in biophysical single-molecule manipulation experiments and accurately tracking their position in 3 dimensions is key for quantitative analysis. Holographic imaging of these beads allows for multiplexing bead tracking but image analysis can be a limiting factor. Here we present a 3D tracking algorithm based on Fast Fourier Transforms that is fast, has nanometric precision, is robust against common artifacts and is accurate over 10's of micrometers. We show its real-time application for magnetic tweezers based force spectroscopy on more than 100 chromatin fibers in parallel, but we anticipate that many other bead-based biophysical essays can benefit from this simple and robust 3 phasor algorithm.

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“…Sound waves can be used to exert forces on objects; this concept is key to the application of controlled forces on surface-tethered microparticles using a method called acoustic force spectroscopy (AFS) [1,14]. By applying force on the microparticle, force is exerted on the tether, DNA or a protein-DNA complex [1,14]. The experimental layout for studying protein-DNA complexes is similar to that used for tethered particle motion (TPM) [2] and magnetic tweezers (MT) (see Chap.…”

Section: Introductionmentioning

confidence: 99%

Unraveling the Biophysical Properties of Chromatin Proteins and DNA Using Acoustic Force Spectroscopy

Lin

Qin

Wuite

et al. 2018

Methods in Molecular Biology

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Acoustic Force Spectroscopy (AFS) is a single-molecule micromanipulation technique that uses sound waves to exert force on surface-tethered DNA molecules in a microfluidic chamber. As large numbers of individual protein-DNA complexes are tracked in parallel, AFS provides insight into the individual properties of such complexes as well as their population averages. In this chapter, we describe in detail how to perform AFS experiments specifically on bare DNA, protein-DNA complexes, and how to extract their (effective) persistence length and contour length from force-extension relations.

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Tuning the Music: Acoustic Force Spectroscopy (AFS) 2.0

Cited by 56 publications

References 28 publications

Single-Cell Acoustic Force Spectroscopy: Resolving Kinetics and Strength of T Cell Adhesion to Fibronectin

Single-Cell Acoustic Force Spectroscopy: Resolving Kinetics and Strength of T Cell Adhesion to Fibronectin

Multiplexed Nanometric 3D Tracking of Microbeads using a FFT-Phasor Algorithm

Unraveling the Biophysical Properties of Chromatin Proteins and DNA Using Acoustic Force Spectroscopy

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