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Dynamic light scattering

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Dynamic light scattering is an analytical technique used to measure the size and distribution of particles in a solution. It works by detecting the Brownian motion of particles and converting this into size information.

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Lab products found in correlation

Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Cup horn sonicator, by Qsonica (1 mentions) Mini extruder, by Avanti Polar Lipids (1 mentions) 1h nmr spectroscopy, by Bruker (1 mentions)

7 protocols using dynamic light scattering

1

Hydrodynamic Radius of PEGylated Proteins

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The Rh of five PEGylated proteins was measured using Dynamic Light Scattering (Wyatt Technology). Proteins in the range of 0.5-1mg/ml were spun down at 13,000rpm for 5 min to pellet aggregates and 20μl was transferred to a quartz cuvette slowly to avoid bubbles. Each sample was measured at least two times, and each time constitutes an average of 30 measurements. Rh values were extracted and plotted against the half-life, cytotoxicity, and anti-tumor activity using GraphPad. Linear regression analysis was performed to obtain a line of best-fit and R2 values.
Zheng Z., Okada R., Kobayashi H., Nagaya T., Wei J., Zhou Q., Lee F., Bera T.K., Gao Y., Kuhlman W., Tai C.H, & Pastan I. (2019). Site-specific PEGylation of anti-mesothelin recombinant immunotoxins increases half-life and anti-tumor activity. Molecular cancer therapeutics, 19(3), 812-821.
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2

Characterizing Lipid Nanoparticle Size

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LNP Z-average hydrodynamic diameter was measured using dynamic light scattering (Wyatt Technologies). LNPs were diluted in sterile 1× PBS to a concentration of ~0.06 μg/ml and analyzed. LNPs were used if they met three criteria: diameter > 20 nm, diameter < 200 nm, and autocorrelation function with only one inflection point. Particles that met these criteria were dialyzed in 1× PBS (Invitrogen) and sterile-filtered with a 0.22-μm filter.
Paunovska K., Da Silva Sanchez A., Foster M.T., Loughrey D., Blanchard E.L., Islam F.Z., Gan Z., Mantalaris A., Santangelo P.J, & Dahlman J.E. (2020). Increased PIP3 activity blocks nanoparticle mRNA delivery. Science Advances, 6(30), eaba5672.
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3

Dynamic Light Scattering of Cationic Liposomes

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The hydrodynamic
size and size distribution of cMLVs were measured by dynamic light
scattering (Wyatt Technology, Santa Barbara, CA).
Liu Y., Fang J., Kim Y.J., Wong M.K, & Wang P. (2014). Codelivery of Doxorubicin and Paclitaxel by Cross-Linked Multilamellar Liposome Enables Synergistic Antitumor Activity. Molecular Pharmaceutics, 11(5), 1651-1661.
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4

Purification and Aggregation of α-Synuclein

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Human monomeric α-Syn (monomer) was purified using size-exclusion chromatography followed by anion exchange. The Pierce high-capacity endotoxin kit was used to remove endotoxins to < 0.1 EU/μg. α-Syn PFFs were generated as described (37 (link)). Monomer concentration was measured using A280 and the extinction coefficient of 5960 M−1cm−1. Monomer was diluted to 5 mg/ml in 150 mM KCl and 50 mM Tris-HCl buffer and shaken for seven days to generate PFFs. After seven days, PFF protein concentration was determined and previously noted buffer was used to bring concentration to 5 mg/ml. On the day of stereotaxic injections, PFFs were sonicated using a cup horn sonicator (QSonica) with a 15°C water bath. Dynamic light scattering (Wyatt Technology) confirmed < 50 nm fragmentation of PFFs. Immediately before injection, 5 mg/ml of monomer was spun at 20,000 × g, and only the supernatant was injected to prevent aggregate formation.
Hong H., Wang Y., Menard M., Buckley J., Zhou L., Volpicelli-Daley L., Standaert D., Qin H, & Benveniste E. (2024). Suppression of the JAK/STAT Pathway Inhibits Neuroinflammation in the Line 61-PFF Mouse Model of Parkinson’s Disease. Research Square.
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5

Preparation and Characterization of DMPG Vesicles

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The stock solution for lipid was prepared by dissolving DMPG in chloroform and then drying under vacuum at ~45 °C for 2 h. The lipid was then mixed with D2O in appropriate amount and vortex mixed for ~10 minutes. The solution then went through four freeze-thaw cycles by alternately placing the suspension in a warm water bath (50 °C) and in a freezer (−80 °C) each for ~30 minutes. The suspension was then vortexed before extruding through a mini-extruder from Avanti Polar Lipids (Alabaster, AL.) fitted with Whatman® Nuclepore™ membranes with an average pore diameter of 100 nm and equilibrated at ~40 °C. The extrusion was performed in sets of 10. Dynamic light scattering (DLS) (Wyatt Technology Corp., Santa Barbara, CA) was used to determine hydrodynamic size distribution and dynamic parameters. DLS clearly indicated uniform vesicle size for both DMPG and DMPG/Aβ vesicles, respectively.
Rai D.K., Sharma V.K., Anunciado D., O’Neill H., Mamontov E., Urban V., Heller W.T, & Qian S. (2016). Neutron Scattering Studies of the Interplay of Amyloid β Peptide(1–40) and An Anionic Lipid 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol. Scientific Reports, 6, 30983.
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6

Nanogel Characterization by GPC and DLS

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The molecular weight and hydrodynamic radius for EHEMA-UDMA, EHEMA-TTEGDMA, and EHEMA-PEG750DMA nanogels were determined using triple-detection gel permeation chromatography (Viscotek) with I-Series column sets (7.8 mm × 30 cm) and linear poly(methyl methacrylate) as an internal standard. Number and weight average molecular weight, hydrodynamic radius, and intrinsic viscosity were collected using in-line refractive index detector, right/low-angle light scattering detector, and capillary viscometer respectively. Sample injections were 100 μL and were eluted at 1 mL/min and 35 °C with THF as solvent. All samples were passed through a 0.45 μm filter prior to injection. Hydrodynamic radius of EHEMA-PEG550DMA nanogels was determined in water using dynamic light scattering (Wyatt Technology) at 20 °C. A sample concentration of 0.2 mg/mL was used and the sample was passed through a 0.45 μm filter prior to analysis. The ratio of EHEMA to UDMA in the isolated nanogel was determined using peak integrals from 1H NMR spectroscopy (Bruker) (400 MHz, CDCl3, EHEMA: δ 3.55–3.80 ppm (m, -CH2CH2O-), UDMA: δ 4.20–4.40 ppm (m, -OCH2CH2O-)).
Dailing E.A., Setterberg W.K., Shah P.K, & Stansbury J.W. (2015). Photopolymerizable Nanogels as Macromolecular Precursors to Covalently Crosslinked Water-Based Networks. Soft matter, 11(28), 5647-5655.
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7

Hydrodynamic Size of cMLVs

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The hydrodynamic size of cMLVs was measured by dynamic light scattering (Wyatt Technology, Santa Barbara, CA). The particles were suspended in filtered water, vortexed and sonicated prior to analysis.
Zhang X., Liu Y., Kim Y.J., Mac J., Zhuang R, & Wang P. (2017). Co-delivery of carboplatin and paclitaxel via cross-linked multilamellar liposomes for ovarian cancer treatment †Electronic supplementary information (ESI) available. See DOI: 10.1039/c7ra01100h Click here for additional data file.. Rsc Advances, 7(32), 19685-19693.
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