Size of Submicrometer Particles Measured by FCS: Correction of the Confocal Volume

Deptuła, Tobiasz; Buitenhuis, Johan; Jarzębski, Maciej; Patkowski, A.; Gapiński, Jacek

doi:10.1021/acs.langmuir.5b01225

Cited by 14 publications

(20 citation statements)

References 14 publications

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“…In our previous publications we have shown that the FCS experiment can be successfully performed at nonstandard conditions, also using long-distance objectives. 2 We have also proposed a method to determine the correct size of uniformly labeled particles comparable to and larger than the confocal volume using FCS, 3 which was in line with the results of other researchers. 4,5 Two recent reviews of FCS applications also deal with that subject.…”

Section: Introductionsupporting

confidence: 79%

“…where a c denotes the core radius. Following the approach presented in our previous study, 3 the interpretation of the current results of both numerical calculations and FCS measurements was initially based on the concept of the effective confocal volume, that is, on the assumption that the product I 0 2 (q) |B(q)| 2 in eq 2 represents an effectively larger (but still Gaussian) beam profile and the correlation functions have the standard mathematical form. With increasing particle size the effective confocal volume becomes less elongated (decreasing fitted c value) because the apparent enlargement is isotropic.…”

mentioning

confidence: 99%

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Structure and Dimensions of Core–Shell Nanoparticles Comparable to the Confocal Volume Studied by Means of Fluorescence Correlation Spectroscopy

Gapiński¹,

Jarzębski

Buitenhuis

et al. 2016

Langmuir

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In some applications the dye distribution within fluorescently labeled nanoparticles and its stability over long periods of time are important issues. In this article we study numerically and experimentally the applicability of fluorescence correlation spectroscopy (FCS) to resolve such questions. When the size of fluorescently labeled particles is comparable to or larger than the confocal volume, the effective confocal volume seen in FCS experiments is increasing. Such an effect has already been studied for uniformly labeled spherical particles. In this work we analyze the form of the FCS correlation functions (CFs) for core-labeled and shell-labeled core-shell particles. For shell-labeled particles an additional fast decay was found both in simulations and in experiments on custom-made surface-labeled particles. Universal scaling of FCS correlation times based on the squared ratio of the labeled part radius of gyration to the Gaussian radius of the beam profile was found. Recipes based on the analysis of simulated CFs, proposed for interpretation of experimental results, were successfully applied to the FCS results on suspensions of large core-labeled and surface-labeled particles.

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Section: Introductionsupporting

confidence: 79%

mentioning

confidence: 99%

Structure and Dimensions of Core–Shell Nanoparticles Comparable to the Confocal Volume Studied by Means of Fluorescence Correlation Spectroscopy

Gapiński¹,

Jarzębski

Buitenhuis

et al. 2016

Langmuir

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“…We applied the core–shell type of nanoparticles to avoid the impact of nanoparticle size on FCS measurements. 32 , 33 Full information on tracers used in the experiments is presented in Supporting Information 1 and 2 ( SI 1, SI 2 ). Cells filled with tracers at final concentrations of 1–100 nM in the cytoplasm were further examined under the confocal microscope.…”

Section: Length-scale Dependent Viscosity Of Cytoplasmmentioning

confidence: 99%

Nanoscale Viscosity of Cytoplasm Is Conserved in Human Cell Lines

Kwapiszewska

Szczepański

Kalwarczyk

et al. 2020

J. Phys. Chem. Lett.

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Metabolic reactions in living cells are limited by diffusion of reagents in the cytoplasm. Any attempt to quantify the kinetics of biochemical reactions in the cytosol should be preceded by careful measurements of the physical properties of the cellular interior. The cytoplasm is a complex, crowded fluid characterized by effective viscosity dependent on its structure at a nanoscopic length scale. In this work, we present and validate the model describing the cytoplasmic nanoviscosity, based on measurements in seven human cell lines, for nanoprobes ranging in diameters from 1 to 150 nm. Irrespective of cell line origin (epithelial–mesenchymal, cancerous–noncancerous, male–female, young–adult), we obtained a similar dependence of the viscosity on the size of the nanoprobes, with characteristic length-scales of 20 ± 11 nm (hydrodynamic radii of major crowders in the cytoplasm) and 4.6 ± 0.7 nm (radii of intercrowder gaps). Moreover, we revealed that the cytoplasm behaves as a liquid for length scales smaller than 100 nm and as a physical gel for larger length scales.

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“…Their unique physical, optical, and electronic features may provide high activity and unusual properties through nanotechnology means [1]. It seems clear that in recent years nanoparticles are popular not only with respect to their potential bioapplications (i.e., drug delivery, biosensors, and bioscaffolds) but also due to the intense development of nanomaterials themselves resulting in numerous model systems as the probes to investigate and/or detect different physical phenomena [2][3][4]. None of the less polymeric nanoparticles have been widely used for biological applications including drug delivery since they must be responsive to external stimuli such as temperature and pH [5].…”

Section: Introductionmentioning

confidence: 99%

pNiPAM-Nanoparticle-Based Antiapoptotic Approach for Pro-Regenerative Capacity of Skeletal Myogenic Cells

et al. 2021

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The biocompatibility of pNiPAM (Poly N-isopropylacrylamide) copolymers has been examined and they did not exert any cytotoxic effects. Their properties and vulnerable temperature characteristics make them candidates for use in medical applications. We synthesized a well-characterized nanoparticles-based cargo system that would effectively deliver a biological agent to human skeletal myogenic cells (SkMCs); among other aspects, a downregulating apoptotic pathway potentially responsible for poor regeneration of myocardium. We confirmed the size of the pNiPAM based spheres at around 100 nm and the nanomeric shape of nanoparticles (NP) obtained. We confirmed that 33 °C is the adequate temperature for phase transition. We performed the dynamics of cargo release. A small amount of examined protein was detected at 10 min after reaching LCTS (lower critical solution temperature). The presented results of the test with BSA (bovine serum albumin) and doxorubicin loaded into nanoparticles showed a similar release profile for both substances. SkMCs incubated with NP loaded with antiapoptotic agent, BCB (Bax channel blocker), significantly diminished cell apoptosis (p < 0.01). Moreover, the lowest apoptotic level was detected in SkMCs treated with camptothecin and simultaneously incubated with pNiPAMs loaded with BCB. Application of nanoparticles loaded with BCB or subjected to BCB alone did not, however, diminish the amount of apparently necrotic cells.

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Size of Submicrometer Particles Measured by FCS: Correction of the Confocal Volume

Cited by 14 publications

References 14 publications

Structure and Dimensions of Core–Shell Nanoparticles Comparable to the Confocal Volume Studied by Means of Fluorescence Correlation Spectroscopy

Structure and Dimensions of Core–Shell Nanoparticles Comparable to the Confocal Volume Studied by Means of Fluorescence Correlation Spectroscopy

Nanoscale Viscosity of Cytoplasm Is Conserved in Human Cell Lines

pNiPAM-Nanoparticle-Based Antiapoptotic Approach for Pro-Regenerative Capacity of Skeletal Myogenic Cells

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