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Syringe pump

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The Syringe Pump is a laboratory instrument designed to accurately and precisely dispense fluids in controlled volumes. It utilizes a motor-driven mechanism to push or pull a syringe plunger, allowing for the delivery of precise amounts of liquid samples or reagents. The Syringe Pump is a versatile tool commonly used in various analytical and experimental procedures that require metered fluid delivery.

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Lab products found in correlation

Scmos camera, by Malvern Panalytical (5 mentions) Nta 3, by Malvern Panalytical (5 mentions) Lm10 nanoparticle analysis system, by Malvern Panalytical (4 mentions) Nanosight ns500, by Malvern Panalytical (2 mentions) Nanosight lm10, by Malvern Panalytical (2 mentions) Nanosight ns300, by Malvern Panalytical (2 mentions)

12 protocols using syringe pump

1

Isolation and Characterization of MSC-Derived Extracellular Vesicles

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WBM or human/murine MSCs were cultured in medium with vesicle depleted FBS for 7 days. Only less than 8 passages of MSCs were used to produce MSC-EVs. The vesicles were isolated from culture medium using differential ultracentrifugation as previously described20 . Unless otherwise noted, all vesicle separations in this study were by differential centrifugation at 300g for 10 minutes, 2,000g for 30 minutes, 10,000g for 1 hour and 100,000g for 1 hour with collection of the 100,000g pellet (exosomes). The vesicles were washed two times with PBS and either tested after storage for 1–7 days at 4°C or resuspended in PBS with 10%DMSO and stored at −80°C. EVs were used within one week after harvested for the in vivo studies. EV functional effects in vitro were maintained for up to 6 months when stored in 10% DMSO at −80°C.
Human and mouse marrow derived MSC-EVs and WBMC-EVs were analyzed by electron microscopy as previously described33 . The pictures are shown in Supplemental Figure 2. Surface epitope protein expression (CD9, CD63 and CD81) in human and mouse marrow derived MSC-EVs and WBMC-EVs were analyzed by Western blot (Supplemental Figure 3 and Table 1). The number and size distribution of vesicles was determined on a NanoSight NS500 (Malvern Instruments, Malvern, UK) with a Syringe Pump (Supplemental Figure 4).
Wen S., Dooner M., Cheng Y., Papa E., Del Tatto M., Pereira M., Deng Y., Goldberg L., Aliotta J., Chatterjee D., Stewart C., Carpanetto A., Collino F., Bruno S., Camussi G, & Quesenberry P. (2016). Mesenchymal stromal cell derived extracellular vesicles rescue radiation damage to murine marrow hematopoietic cells. Leukemia, 30(11), 2221-2231.
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2

Nanoparticle Tracking Analysis of Fungal EVs

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Nanoparticle tracking analysis (NTA) of fungal EVs was performed on an LM10 nanoparticle analysis system, coupled with a 488-nm laser and equipped with a scientific complementary metal oxide semiconductor (SCMOS) camera and a syringe pump (Malvern Panalytical, Malvern, UK) following the conditions used by Reis and colleagues (46 (link)). All samples were 833- to 2,500-fold diluted in filtered PBS and measured within the optimal dilution range of 9 × 107 to 2.9 × 109 particles/mL. Samples were injected using a syringe pump speed of 50, and three videos of 60 s were captured per sample, with the camera level set to 15, gain set to 3, and viscosity set to that of water (0.954 to 0.955 cP). For data analysis, the gain was set to 10 to 15, and the detection threshold was set to 2 to 3 for all samples. Levels of blur and maximum jump distance were automatically set. The data were acquired and analyzed using the NTA 3.0 software (Malvern Panalytical).
Honorato L., de Araujo J.F., Ellis C.C., Piffer A.C., Pereira Y., Frases S., de Sousa Araújo G.R., Pontes B., Mendes M.T., Pereira M.D., Guimarães A.J., da Silva N.M., Vargas G., Joffe L., Del Poeta M., Nosanchuk J.D., Zamith-Miranda D., dos Reis F.C., de Oliveira H.C., Rodrigues M.L., de Toledo Martins S., Alves L.R., Almeida I.C, & Nimrichter L. (2022). Extracellular Vesicles Regulate Biofilm Formation and Yeast-to-Hypha Differentiation in Candida albicans. mBio, 13(3), e00301-22.
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3

Nanoparticle Tracking Analysis of Calcific EV Aggregation

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NTA by NanoSight LM10 (Malvern Instruments Ltd, Malvern, UK) assessed vesicle size and concentration within the media samples46 (link). Calcific SMC-derived EVs collected from conditioned media by ultracentrifugation at 100,000 × g were resuspended in 1 ml calcifying media and divided into five 200 μl portions. One portion was collected for initial time point analyses. Aggregation was detected by NTA in the remaining portions incubated at 37 °C for 1, 3, 5, or 7 days. NTA determines nanoparticle number and size distributions from the Brownian motions of objects illuminated by laser light47 (link). The samples were diluted 1:7 in PBS prior to injection into the NanoSight chamber. Samples were continuously injected via a syringe pump (Malvern), and five NTA videos were collected for 1 min each. The camera gain was set at a constant value of 9, and the threshold value for vesicle detection was set to 2. The resultant size and concentration output data from each video were averaged to generate the final distribution for each sample.
Hutcheson J.D., Goettsch C., Bertazzo S., Maldonado N., Ruiz J.L., Goh W., Yabusaki K., Faits T., Bouten C., Franck G., Quillard T., Libby P., Aikawa M., Weinbaum S, & Aikawa E. (2016). Genesis and growth of extracellular vesicle-derived microcalcification in atherosclerotic plaques. Nature materials, 15(3), 335-343.
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4

Extracellular Vesicle Size Analysis

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Nanoparticle Tracking Analysis (NTA) was performed on a NanoSight NS300 (Malvern Panalytical, Malvern, UK) at RT. EV concentrates obtained post-SEC were diluted 1:10 in Hepes buffer, then underwent bath sonication in a Branson 2510 bath sonicator (Branson, Danbury CT) for 30 seconds at RT to reduce sample aggregation. EVs were then diluted (1:1,000 to 1:10,000 depending on sample) and added to a 1 mL syringe, then set on a syringe pump (Malvern Panalytical) and loaded into the NanoSight low volume flow cell. Each sample was analyzed using a 405 nm laser with 5 consecutive 1 minute video recordings with a constant flow rate set at 10 (no units), flow rates are set in the software and do not contain units. Videos were compiled and analyzed in the NTA software (Version 3.4). All videos were compiled and analyzed together in the NTA software and data were collected and saved in raw form.
Marsh S.R., Pridham K.J., Jourdan J, & Gourdie R.G. (2021). Novel Protocols for Scalable Production of High Quality Purified Small Extracellular Vesicles from Bovine Milk. Nanotheranostics, 5(4), 488-498.
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5

Nanoparticle Tracking Analysis of Extracellular Vesicles

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The method used for monitoring the presence of EVs in our samples was the nanoparticle tracking analysis (NTA). This analysis was performed on an LM10 nanoparticle analysis system, coupled with a 488-nm laser and equipped with an SCMOS camera and a syringe pump (Malvern Panalytical, Malvern, United Kingdom). All samples were measured according to the conditions previously described by our group (2 (link)). The data were acquired and analyzed using the NTA 3.0 software (Malvern Panalytical). Particle numbers obtained by NTA were normalized to the number of cells in the cultures for determination of the EV/cell ratios. NTA histograms were also used for the quantification of EV peaks in different samples using the Image J software (https://imagej.nih.gov/ij/). In the gradient samples, diameter ranges corresponded to 300 to 400 nm, 400 to 600 nm, and 600 to 900 nm. In the kinetics studies, EV diameters were simply separated into 0 to 200 nm and larger than 200 nm. In both assays, these regions of the histograms were manually colored using the Image J paintbrush tool, and their areas were automatically measured using the wand/tracing tool.
Reis F.C., Gimenez B., Jozefowicz L.J., Castelli R.F., Martins S.T., Alves L.R., de Oliveira H.C, & Rodrigues M.L. (2021). Analysis of Cryptococcal Extracellular Vesicles: Experimental Approaches for Studying Their Diversity Among Multiple Isolates, Kinetics of Production, Methods of Separation, and Detection in Cultures of Titan Cells. Microbiology Spectrum, 9(1), 10.1128/spectrum.00125-21.
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6

Nanoparticle Analysis of Extracellular Vesicles

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EVs were analysed using the NanoSight NS300 (Malvern Instrument, Westborough, MA) for size, shape and concentration. NanoSight employs laser illumination to provoke light scattering and video capture of Brownian motion to determine the size distribution and concentration of particles in solution. EVs (diluted 1:250; ~107–109 particles/mL) were injected with a syringe pump (infuse = 20, Malvern Instrument) and recorded (3 × 30-second captures) to determine the mean size and concentration of nanoparticles in each sample.
Flemming J.P., Hill B.L., Haque M.W., Raad J., Bonder C.S., Harshyne L.A., Rodeck U., Luginbuhl A., Wahl JK I.I.I., Tsai K.Y., Wermuth P.J., Overmiller A.M, & Mahoney M.G. (2020). miRNA- and cytokine-associated extracellular vesicles mediate squamous cell carcinomas. Journal of Extracellular Vesicles, 9(1), 1790159.
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7

Extracellular Vesicle Isolation and Characterization from Yeast Cultures

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Parental and mutant strains were cultivated in yeast extract–peptone–dextrose (YPD, 5 ml) medium for 24 h (30°C, with shaking). The cell suspensions were adjusted to 3.5 × 107 cells/ml and 300 μl were inoculated onto YPD agar plates (n = 3). The cultures were incubated to confluence for 24 h at 30°C.
The cells were gently recovered from each plate with an inoculation loop and suspended in PBS (30 ml). For removal of the cells and debris, the suspensions were first centrifuged at 5000 × g for 15 min at 4°C, and the resulting supernatants were centrifuged at 15 000 × g for 15 min at 4°C. The supernatants were filtered through 0.45-μm pore syringe filters and centrifuged at 100 000 × g for 1 h at 4°C to recover EVs. EVs were analyzed by nanoparticle tracking analysis (NTA) on an LM10 nanoparticle analysis system, coupled with a 488-nm laser and equipped with an SCMOS camera and a syringe pump (Malvern Panalytical, Malvern, United Kingdom), as previously published (Reis et al. 2021 (link)). Data were acquired and analyzed using the NTA 3.0 software (Malvern Panalytical).
Jung E.H., Park Y.D., Dragotakes Q., Ramirez L.S., Smith D.Q., Reis F.C., Dziedzic A., Rodrigues M.L., Baker R.P., Williamson P.R., Jedlicka A., Casadevall A, & Coelho C. (2022). Cryptococcus neoformans releases proteins during intracellular residence that affect the outcome of the fungal–macrophage interaction. microLife, 3, uqac015.
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8

Nanoparticle Tracking Analysis of Calcific EV Aggregation

Cited 1 time
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NTA by NanoSight LM10 (Malvern Instruments Ltd, Malvern, UK) assessed vesicle size and concentration within the media samples46 (link). Calcific SMC-derived EVs collected from conditioned media by ultracentrifugation at 100,000 × g were resuspended in 1 ml calcifying media and divided into five 200 μl portions. One portion was collected for initial time point analyses. Aggregation was detected by NTA in the remaining portions incubated at 37 °C for 1, 3, 5, or 7 days. NTA determines nanoparticle number and size distributions from the Brownian motions of objects illuminated by laser light47 (link). The samples were diluted 1:7 in PBS prior to injection into the NanoSight chamber. Samples were continuously injected via a syringe pump (Malvern), and five NTA videos were collected for 1 min each. The camera gain was set at a constant value of 9, and the threshold value for vesicle detection was set to 2. The resultant size and concentration output data from each video were averaged to generate the final distribution for each sample.
Hutcheson J.D., Goettsch C., Bertazzo S., Maldonado N., Ruiz J.L., Goh W., Yabusaki K., Faits T., Bouten C., Franck G., Quillard T., Libby P., Aikawa M., Weinbaum S, & Aikawa E. (2016). Genesis and growth of extracellular vesicle-derived microcalcification in atherosclerotic plaques. Nature materials, 15(3), 335-343.
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9

Nanoparticle Tracking Analysis of Extracellular Vesicles

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The method used for monitoring the presence of EVs in our samples was the nanoparticle tracking analysis (NTA). This analysis was performed on an LM10 nanoparticle analysis system, coupled with a 488-nm laser and equipped with an SCMOS camera and a syringe pump (Malvern Panalytical, Malvern, United Kingdom). All samples were measured according to the conditions previously described by our group (2 (link)). The data were acquired and analyzed using the NTA 3.0 software (Malvern Panalytical). Particle numbers obtained by NTA were normalized to the number of cells in the cultures for determination of the EV/cell ratios. NTA histograms were also used for the quantification of EV peaks in different samples using the Image J software (https://imagej.nih.gov/ij/). In the gradient samples, diameter ranges corresponded to 300 to 400 nm, 400 to 600 nm, and 600 to 900 nm. In the kinetics studies, EV diameters were simply separated into 0 to 200 nm and larger than 200 nm. In both assays, these regions of the histograms were manually colored using the Image J paintbrush tool, and their areas were automatically measured using the wand/tracing tool.
Reis F.C., Gimenez B., Jozefowicz L.J., Castelli R.F., Martins S.T., Alves L.R., de Oliveira H.C, & Rodrigues M.L. (2021). Analysis of Cryptococcal Extracellular Vesicles: Experimental Approaches for Studying Their Diversity Among Multiple Isolates, Kinetics of Production, Methods of Separation, and Detection in Cultures of Titan Cells. Microbiology Spectrum, 9(1), 10.1128/spectrum.00125-21.
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10

Isolation and Characterization of MSC-Derived Extracellular Vesicles

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WBM or human/murine MSCs were cultured in medium with vesicle depleted FBS for 7 days. Only less than 8 passages of MSCs were used to produce MSC-EVs. The vesicles were isolated from culture medium using differential ultracentrifugation as previously described20 . Unless otherwise noted, all vesicle separations in this study were by differential centrifugation at 300g for 10 minutes, 2,000g for 30 minutes, 10,000g for 1 hour and 100,000g for 1 hour with collection of the 100,000g pellet (exosomes). The vesicles were washed two times with PBS and either tested after storage for 1–7 days at 4°C or resuspended in PBS with 10%DMSO and stored at −80°C. EVs were used within one week after harvested for the in vivo studies. EV functional effects in vitro were maintained for up to 6 months when stored in 10% DMSO at −80°C.
Human and mouse marrow derived MSC-EVs and WBMC-EVs were analyzed by electron microscopy as previously described33 . The pictures are shown in Supplemental Figure 2. Surface epitope protein expression (CD9, CD63 and CD81) in human and mouse marrow derived MSC-EVs and WBMC-EVs were analyzed by Western blot (Supplemental Figure 3 and Table 1). The number and size distribution of vesicles was determined on a NanoSight NS500 (Malvern Instruments, Malvern, UK) with a Syringe Pump (Supplemental Figure 4).
Wen S., Dooner M., Cheng Y., Papa E., Del Tatto M., Pereira M., Deng Y., Goldberg L., Aliotta J., Chatterjee D., Stewart C., Carpanetto A., Collino F., Bruno S., Camussi G, & Quesenberry P. (2016). Mesenchymal stromal cell derived extracellular vesicles rescue radiation damage to murine marrow hematopoietic cells. Leukemia, 30(11), 2221-2231.
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