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Optical tweezers

Learn all about optical tweezers and what they can do

What are optical tweezers and what can they do?

In 2018 Arthur Ashkin won the Nobel Prize in Physics “for the optical tweezers and their application to biological systems”. In essence, he discovered that light’s momentum could serve as an incredibly sensitive set of tweezers to catch and study biomolecules.

By shooting a laser through a microscope, he created a highly focused light beam, strong enough to trap and “trap” small objects, such as plastic beads. These beads can be coated to stick to various biomolecules, such as DNA, RNA, proteins, or filaments. Two trapped beads can be used as hooks to hold a single molecule on its respective ends. We can then control the lasers and move the beads with the tethered biomolecule and, for instance, stretch it. The ability to manipulate molecules in this way allows us to measure forces (for example, upon stretching) and monitor the beads’ positions. These outcomes can then serve us to calculate the tethered molecule (for example, its elasticity) and study structural transitions.

Discovery of optical tweezers

First introduced by Arthur Ashkin and colleagues in 1986 at Bell labs, optical tweezers quickly emerged as an indispensable tool that can be used for a variety of different applications in chemistry and biology1.

Not long after this initial breakthrough, optical tweezers, or optical traps as they are otherwise known, were successfully used to physically trap and control viruses, bacteria and single-cells, paving the way for the mechanical and kinetic study of biomolecules at the single-molecule level2-3.

Presently, optical tweezers have been used in a variety of different applications, such as in the study of the interactions between proteins and DNA involved in DNA organization, replication, transcription and repair; the study of the energy landscape of proteins and the kinetics of molecular motors4-7.

Simply put, the value of optical tweezers lies in the fact that they can be used to perform experiments to probe the properties of single-molecules by applying forces in the range of picoNewtons and by measuring distance displacements in the range of nanometers.
Artistic sketch of a Arthur Ashkin, Nobel Prize winner by single-molecule trap inviation: a smiling man with a beard and short hair in simple black and gold strokes.

Working principle of optical tweezers

Optical tweezers are based on the principle of light carrying momentum proportional to its energy and propagation direction.

When a laser beam passes through an object, it bends and changes direction (called refraction) and alters its momentum. According to Newton’s third law, the object undergoes an equal and opposite momentum change, a reaction force, for the system to conserve the total momentum.

Figure 1 illustrates the transfer of light momentum occurring when a light beam travels through a bead. In a typical optical tweezers configuration, the incoming light originates from a focused laser beam through a microscope objective and focuses on a spot in the sample. The spot subsequently creates a trap able to hold a small object in place.
Diagram showing light path refraction and momentum change passing through a bead, bending downward and to the side.
Figure 1: Re-direction of a light path and change of momentum as it passes through a microsphere or “bead” with high index of refraction related to the medium (left). Momentum of equal and opposite force is transferred from the photons to the bead according to Newton’s Law of energy conservation (right).

Laser trapping of beads

In a typical optical tweezers configuration, the incoming light originates from a focused laser beam through a microscope objective and focuses on a spot in the sample. The spot creates a trap able to hold a small dielectric object at place.

The total forces experienced by the object, or bead in most experimental settings, consist of a scattering force and a gradient force8. The scattering force arises when a light beam is scattered by the surface of the object. This scattering produces a net momentum transfer from the light photons to the object and causes the bead to be pushed towards the beam propagation. The gradient force results from the intensity profile of the laser beam which acts as an attractive force, drawing the bead towards the region with greater light intensity. In the case of a focused laser beam with a Gaussian intensity profile (a normal distribution), the gradient force pulls the object into the center of the focal plane.

The reason the object stays in the center of the beam is because of the sum of the forces acting upon it. In the center, rays of light refract or scatter through the object the same way on both sides of the vertical plane, which cancels forces from moving the object sideways. If the object drifts to one side, it returns to the center. Think of a spring that accelerates back to the center when displaced from its equilibrium position. Figure 2 shows how the gradient force restores an off-centered bead towards the center of the focal plane, eff ectively trapping the object in all dimensions
Figure 2: Two light paths passing through a dielectric micron sized bead. Due to the light gradient, the path originating from the center of the beam carries more photons than the light path commencing from the outlines of the beam, resulting to a larger force pulling the bead towards the focal point.

References

1. Ashkin, A. et al.
1986
Optic Letters
https://doi.org/10.1364/OL.11.000288
1. Ashkin, A. et al.
2. Ashkin, A. et al.
1987
Science
https://doi.org/10.1126/science.3547653
2. Ashkin, A. et al.
3. Ashkin, A. et al.
1987
Nature
https://doi.org/10.1038/330769a0
3. Ashkin, A. et al.
4. Dame, R. et al.
2006
Nature
https://doi.org/10.1038/nature05283
4. Dame, R. et al.
5. Heller, I. et al.
2014
Chemical Reviews
https://doi.org/10.1021/cr4003006
5. Heller, I. et al.
6. Cecconi, C. et al.
2005
Science
https://doi.org/10.1126/science.1116702
6. Cecconi, C. et al.
7. Nishizaka, T. et al.
1995
Nature
https://doi.org/10.1038/377251a0
7. Nishizaka, T. et al.
8. Neuman, K. et al.
2004
Review of Scientific Instruments
https://doi.org/10.1063/1.1785844
8. Neuman, K. et al.
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The CAR Immune Synapse Interactome: How Membrane Organization Shapes CAR T–Tumor Cell Interactions
The CAR Immune Synapse Interactome: How Membrane Organization Shapes CAR T–Tumor Cell Interactions
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Unlike conventional drugs, whose activity is shaped by binding to a single target, CAR T-cell (CART) engagement with tumor cells involves multiple receptors, adhesion molecules, and signaling proteins. These components are partially organized within cholesterol-rich membrane microdomains called lipid rafts, forming the CAR immune synapse (CARIS)—the interface through which a CART recognizes, attaches to, and kills a tumor cell.

To evaluate CARIS as a pharmacological interface, we compared CD28- and 4-1BB-CARTs, which exhibit distinct therapeutic profiles. The rapid antitumor activity of CD28-CARTs was associated with brief, dynamic synapses supporting rapid signaling and serial killing. Conversely, 4-1BB-CARTs formed longer, mechanically stable synapses consistent with their slower but sustained activity. Synapse strength was measured using z-Movi acoustic force microscopy.

Lipid rafts were enriched at CARIS. Attenuating membrane lipid raft cholesterol preferentially compromised 4-1BB CARIS mechanical stability, while predominantly impairing CD28 CARIS activation and cytolytic output. The molecular networks underlying these differences were mapped by integrating lipid-raft lipidomics, differential SILAC proteomics to distinguish CART-derived from tumor-derived components, and T-cell transcriptomics.

CD28-associated lipid rafts were enriched in diacylglycerol signaling components, PKCθ, small GTPases, Src-family kinases, and AKT/PI3K-proximal modules, supporting rapid signal-to-function coupling. Stomatin-family and flotillin scaffolds favored dynamic remodeling, while CD8-dominant transcriptional programs supported cytolytic degranulation. Engaged tumor cells displayed membrane-stress signatures consistent with late-stage senescence and apoptotic commitment.

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Accelerate your cell engager discovery with high throughput measurements of Cell Avidity
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T cells play a pivotal role in tumor immunosurveillance. Multispecific cell engagers (CEs) have been adopted in the field of immuno-oncology to redirect T cells toward cancer cells, thereby unleashing the anti-tumor potential of the patient’s immune system. CE-mediated cell binding induces T cell activation and the formation of an immunological synapse, which is a prerequisite for effective tumor cell lysis.

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