Nanosphere size standard

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom

The Nanosphere size standard is a reference material for calibrating and verifying the performance of instruments used to measure the size of nanoparticles. It consists of a suspension of uniform polystyrene microspheres with a known diameter, which can be used to validate the accuracy and precision of nanoparticle size measurement techniques.

Automatically generated - may contain errors

Lab products found in correlation

Zetaview nanoparticle tracker, by Particle Metrix (2 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Dynapro, by Wyatt Technology (1 mentions) Ns300, by Malvern Panalytical (1 mentions) Zetasizer nano zs90, by Malvern Panalytical (1 mentions)

Close spelling variants of this product

Nanosphere (2 citations) 3000 Series Nanosphere Size Standards (2 citations) 3100A Nanosphere Size Standard (2 citations)

7 protocols using nanosphere size standard

Quantifying EV Size and Concentration

Check if the same lab product or an alternative is used in the 5 most similar protocols

Size and number of EVs were quantified using a ZetaView Nanoparticle Tracker (Particle Metrix GmBH, Meerbusch, Germany), and corresponding ZetaVeiw software (8.03.04.01). A nanosphere size standard (100 nm diameter; Thermo Scientific; Waltham, MA) was used to calibrate the instrument prior to readings. Instrument pre-acquisition parameters were set to a temperature of 23°C, sensitivity of 65, frame rate of 30 frames per second (fps), shutter speed of 100, and a laser pulse duration equal to that of shutter duration. Post-acquisition parameters were set to a minimum brightness of 25, maximum size of 200 pixels, and a minimum size of 10 pixels. For each sample 1 mL of diluted EVs were injected into the sample-carrier cell and the particle count was measured at five positions, with two cycles of reading per position. The cell was washed with PBS after every sample. The mean size and concentration of EVs/mL (±SEM) was calculated from four replicate experiments. The coefficient of variation (CV) as determined from a pooled sample was 4.8% for size and 4.4% for concentration of EVs.

Chen S., Datta-Chaudhuri A., Deme P., Dickens A., Dastgheyb R., Bhargava P., Bi H, & Haughey N.J. (2019). Lipidomic characterization of extracellular vesicles in human serum. Journal of Circulating Biomarkers, 8, 1849454419879848.

+ Open protocol

+ Expand

Quantifying Extracellular Vesicle Size and Concentration

Check if the same lab product or an alternative is used in the 5 most similar protocols

EV size and quantity were determined using ZetaView Nanoparticle Tracker (Particle Metrix, Cambridge, UK) with the ZetaView software (8.04.02 SP2). The calibration of the instrument was performed using a nanosphere size standard (100 nm diameter; Thermo Fisher Scientific, Abingdon, UK). Instrument pre-acquisition parameters were set to a temperature of 23 °C, sensitivity of 80, frame rate of 30 frames per second (fps), shutter speed of 100, and a laser pulse duration equal to that of shutter duration. Post-acquisition parameters were set to a minimum brightness of 25, maximum size of 1000 pixels, and a minimum size of 5 pixels. Each sample was diluted 1:3000 and 1 mL of diluted EVs was injected into the sample-carrier cell and the particle count was measured at eleven positions, with two cycles of reading per position. Samples were read in duplicate. The cell was washed with PBS (Gibco, Abingdon, UK) after every sample. The mean size and concentration of EVs/mL (±SEM) was calculated for each group.

Arrifano G.D., Augusto-Oliveira M., Sealey-Bright M., Zainal J., Imbiriba L., Fernandes L.M., Maia C.S., Anthony D, & Crespo-Lopez M.E. (2021). Contributing to Understand the Crosstalk between Brain and Periphery in Methylmercury Intoxication: Neurotoxicity and Extracellular Vesicles. International Journal of Molecular Sciences, 22(19), 10855.

+ Open protocol

+ Expand

Nanosphere Size Standard Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols

A sample of Nanosphere^™ size standard polystyrene spheres (1% w/v in aqueous suspension) with a calibrated mean diameter of 101 nm ± 3 nm and a standard deviation of 6.2 nm was obtained from the manufacturer (Thermo Scientific, catalog #: 3100/3100A) and diluted by a factor of 500 into 0.5% aqueous acetic acid. The calibrated mean diameter (certified batch #: 3100–009) is certified by the manufacturer via transfer by TEM from National Institute of Standards and Technology certified microspheres. A ~50 nm surface amine-functionalized polystyrene sphere sample (≥2.5% w/v aqueous suspension) was obtained from the manufacturer (Sigma Aldrich, product #: L0780) and diluted by a factor of 5,000 into 0.5% acetic acid. The manufacturer reported mean diameter range for this sample is 45–55 nm. The density of polystyrene is 1.0502 g/mL;⁴⁰ this value is used in all calculations of diameter determined from measured masses.

Balasundaram D., Tabor C.W, & Tabor H. (1999). Sensitivity of Spermidine-Deficient Saccharomyces cerevisiae to Paromomycin. Antimicrobial Agents and Chemotherapy, 43(5), 1314-1316.

+ Open protocol

+ Expand

Quantifying Size and Number of ADEVs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Size and number of ADEVs were quantified using a ZetaView Nanoparticle Tracker (Particle Metrix GmBH, Meerbusch, Germany), and corresponding ZetaVeiw software (8.03.04.01). A nanosphere size standard (100 nm diameter; Thermo Scientific) was used to calibrate the instrument prior to readings. Instrument pre-acquisition parameters were set to a temperature of 23 °C, sensitivity of 65, frame rate of 30 frames per second (fps), shutter speed of 100, and a laser pulse duration equal to that of shutter duration. Post-acquisition parameters were set to a minimum brightness of 25, maximum size of 200 pixels, and a minimum size of 10 pixels. For each sample 1 mL of diluted EVs were injected into the sample-carrier cell and the particle count was measured at five positions, with two cycles of reading per position. The cell was washed with PBS after every sample. The mean size and concentration of EVs/mL (±SEM) was calculated from four replicate experiments. The coefficient of variation as determined from a pooled sample was 4.8% for size and 4.4% for concentration of EVs.

Chaudhuri A.D., Dastgheyb R.M., Yoo S.W., Trout A., Talbot Jr C.C., Hao H., Witwer K.W, & Haughey N.J. (2018). TNFα and IL-1β modify the miRNA cargo of astrocyte shed extracellular vesicles to regulate neurotrophic signaling in neurons. Cell Death & Disease, 9(3), 363.

+ Open protocol

+ Expand

Phosphatidylserine Liposomes Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Phosphatidylserine (PS) liposomes were prepared by suspending the lipid (porcine brain; Avanti Polar Lipids #840032P) at 5 mg/ml (13 mM) in 50 mM Tris buffer, pH 7.5. The suspension was extruded 11 times through a 50 nm pore polycarbonate membrane using an Avestin LiposoFast-Basic Membrane Extruder and the solution was stored at 4°C. This method is similar to previously described methods for the preparation of phosphatidylcholine-based liposomes containing PS [52 (link)] with the exception that PS was the only lipid solubilized and it was dissolved directly in 50mM Tris pH 7.5 prior to extrusion. The resulting liposomes were characterized in triplicate by dynamic light scattering on a Wyatt DynaPro reader and were 120nm (polydispersity 16nm). The expected sizes for 100 and 300nm Nanosphere^™ size standards (ThermoFisher Scientific 3100A and 3300A, respectively) were observed. Upon addition of KRAS, the liposomes were 129nm (polydispersity 19nm) in the presence of assay buffer.

Jansen J.M., Wartchow C., Jahnke W., Fong S., Tsang T., Pfister K., Zavorotinskaya T., Bussiere D., Cheng J.M., Crawford K., Dai Y., Dove J., Fang E., Feng Y., Florent J.M., Fuller J., Gossert A.D., Hekmat-Nejad M., Henry C., Klopp J., Lenahan W.P., Lingel A., Ma S., Meyer A., Mishina Y., Narberes J., Pardee G., Ramurthy S., Rieffel S., Stuart D., Subramanian S., Tandeske L., Widger S., Widmer A., Winterhalter A., Zaror I, & Hardy S. (2017). Inhibition of prenylated KRAS in a lipid environment. PLoS ONE, 12(4), e0174706.

+ Open protocol

+ Expand

Nanoparticle Characterization using NTA

Check if the same lab product or an alternative is used in the 5 most similar protocols

Nanoparticle tracking analysis (NTA) was performed using a Malvern NanoSight NS300 instrument. Samples and standards were diluted in 0.1 μM filtered and degassed PBS. Nanosphere size standards (125nM, Thermo) were diluted to 1:5,000 and 1:10,000 and used to perform initial focus for the NTA reading, followed by washing with degassed PBS. After each sample was processed, the lines were thoroughly washed again with PBS. Parameters were set as follows: flow rate of 50 μl/min, with five 30-s videos recorded for analysis.

O’Hara B.A., Morris-Love J., Gee G.V., Haley S.A, & Atwood W.J. (2020). JC Virus infected choroid plexus epithelial cells produce extracellular vesicles that infect glial cells independently of the virus attachment receptor. PLoS Pathogens, 16(3), e1008371.

+ Open protocol

+ Expand

Measuring Astrocyte-Derived EV Size

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

To measure the size of the EVs released from astrocyte processes we performed the nanosight analysis on ultracentrifugation pellet resuspended in PBS using the Zetasizer Nano ZS90 particle sizer at a 90° fixed angle (Malvern Instruments, Worcestershire, United Kingdom), as previously described (Marimpietri et al., 2013 (link)). Nanosphere™ size standards with a mean diameter of 57 ± 4 nm (Thermo Scientific) were used for particle sizer calibration. The analysis was replicated on three different samples.

Venturini A., Passalacqua M., Pelassa S., Pastorino F., Tedesco M., Cortese K., Gagliani M.C., Leo G., Maura G., Guidolin D., Agnati L.F., Marcoli M, & Cervetto C. (2019). Exosomes From Astrocyte Processes: Signaling to Neurons. Frontiers in Pharmacology, 10, 1452.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!