Real‐Time Feedback‐Driven Single‐Particle Tracking: A Survey and Perspective

Heerden, Bertus van; Vickers, Nicholas A.; Krüger, Tjaart P. J.; Andersson, Sean B.

doi:10.1002/smll.202206874

Cited by 1 publication

(2 citation statements)

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“…Even longer trajectories could be anticipated using a feedback loop to keep the particle of interest in the detection volume. 1,57 This will provide information on the long-term dynamics of the targeted components, such as, for instance, the accumulation of receptors in particular domains on the plasma membrane. In principle, it should be possible to track the same FcεRI receptor before and after its antigen-induced clustering with other receptors until its recycling and internalization by the cell.…”

Section: Discussionmentioning

confidence: 99%

“…As a control, and to validate our microscopy setup and our analysis, we tracked the Brownian diffusion of polymer-coated UCNPs in water−glycerol mixtures of known viscosity. 57 The protocol and results of the experiment are given in Figure S12. Moreover, we also assessed possible heating effects induced by laser illumination in our microscope setup by monitoring the Brownian diffusion of coated UCNPs in water−glycerol mixtures for different laser intensities (Figure S12).…”

Section: Spt Of Ucnp-labeled Fcεri Receptors On Living Mast Cellsmentioning

confidence: 99%

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Targeted Single Particle Tracking with Upconverting Nanoparticles

Dukhno,

Ghosh,

Greiner

et al. 2024

ACS Appl. Mater. Interfaces

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Single particle tracking (SPT) is a powerful technique for real-time microscopic visualization of the movement of individual biomolecules within or on the surface of living cells. However, SPT often suffers from the suboptimal performance of the photon-emitting labels used to tag the biomolecules of interest. For example, fluorescent dyes have poor photostability, while quantum dots suffer from blinking that hampers track acquisition and interpretation. Upconverting nanoparticles (UCNPs) have recently emerged as a promising anti-Stokes luminescent label for SPT. In this work, we demonstrated targeted SPT using UCNPs. For this, we synthesized 30 nm diameter doped UCNPs and coated them with amphiphilic polymers decorated with polyethylene glycol chains to make them waterdispersible and minimize their nonspecific interactions with cells. Coated UCNPs highly homogeneous in brightness (as confirmed by a single particle investigation) were functionalized by immunoglobulin E (IgE) using a biotin−streptavidin strategy. Using these IgE-UCNP SPT labels, we tracked high-affinity IgE receptors (FcεRI) on the membrane of living RBL-2H3 mast cells at 37 °C in the presence and absence of antigen and obtained good agreement with the literature. Moreover, we used the FcεRI-IgE receptorantibody system to directly compare the performance of UCNP-based SPT labels to organic dyes (AlexaFluor647) and quantum dots (QD655). Due to their photostability as well as their backgroundless and continuous luminescence, SPT trajectories obtained with UCNP labels are no longer limited by the photophysics of the label but only by the dynamics of the system and, in particular, the movement of the label out of the field of view and/or focal plane.

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Section: Discussionmentioning

confidence: 99%

Section: Spt Of Ucnp-labeled Fcεri Receptors On Living Mast Cellsmentioning

confidence: 99%