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Avanti mini extruder

Manufactured by Cytiva
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Sourced in United Kingdom

The Avanti mini extruder is a compact and versatile laboratory instrument designed for extrusion of small sample sizes. It features a pneumatically driven piston that applies controlled pressure to a sample, enabling the extrusion of materials through a variety of interchangeable dies. The Avanti mini extruder is a practical tool for researchers and scientists requiring precise control over sample extrusion in a laboratory setting.

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Lab products found in correlation

Polycarbonate membrane, by Cytiva (2 mentions) Polycarbonate filter, by Cytiva (1 mentions) Rotary evaporator, by Heidolph (1 mentions) Fs30d bath sonicator, by Thermo Fisher Scientific (1 mentions) Nanosight ns300, by Malvern Panalytical (1 mentions)

6 protocols using avanti mini extruder

1

Preparation of Supported Lipid Bilayers

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Membrane compositions used in this study were DOPC:DOPS:PI(4,5)P2 at 80:15:5 or DOPC:DOPS:PI(4,5)P2:Rh-PE at 79:15:5:1 for fluorescent-tagged membrane. For liposome preparation, lipid mixtures were dried, rehydrated in buffer containing 20 mM Hepes (pH 7.5) and 150 mM KCl, and then subjected to three rapid freeze–thaw cycles followed by extrusion through a 0.1-µm polycarbonate membrane (Whatman) using Avanti mini extruder. SUPER templates were generated by incubating 2.5-µm silica beads in a solution containing 100 µM fluorescent-tagged liposomes and 1 M NaCl for 30 min at room temperature with intermittent flicking. Excess unbound liposomes were washed four times with filtered water after incubation (Neumann et al., 2013 (link)).
Laiman J., Hsu Y.J., Loh J., Tang W.C., Chuang M.C., Liu H.K., Yang W.S., Chen B.C., Chuang L.M., Chang Y.C, & Liu Y.W. (2022). GSK3α phosphorylates dynamin-2 to promote GLUT4 endocytosis in muscle cells. The Journal of Cell Biology, 222(2), e202102119.
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2

Liposome Preparation and Copolymer Incorporation

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Liposomes were prepared using the film rehydration and extrusion procedure, and copolymer incorporation was accomplished via the postinsertion mechanism. In general, a desired amount of DPPC was dissolved in CHCl3, which was subsequently removed using a steady stream of ultrahigh-purity N2, forming a thin, solid film. The films were further dried under vacuum at 55 °C for 18 h. For rehydration, H2O or D2O was added to the films, and they were held at 55 °C with periodic agitation over the period of an hour followed by aging at 55 °C overnight. The resulting dispersions were extruded through 400 nm (20 passes), 200 nm (20 passes), and 100 nm (41 passes) polycarbonate filters (Whatman) using an Avanti Mini-Extruder held at 55 °C. The postinsertion of PEG-P(8C-Chol) was carried out by mixing extruded DPPC liposomes with the appropriate amount of separately prepared aqueous polymer solution. For the most part, the precursor copolymer solutions were prepared by direct dissolution in water using a combination of mixing and sonication. The only exception I s PEGP(8C-Chol)4.8, which required dialysis from dimethylformamide (DMF). Following initial mixing, the formulations were maintained at 55 °C for 1 h with regular agitation.
Mineart K.P., Venkataraman S., Yang Y.Y., Hedrick J.L, & Prabhu V.M. (2018). Fabrication and Characterization of Hybrid Stealth Liposomes. Macromolecules, 51(8), 3184-3192.
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3

Liposome Preparation by Thin-Film Hydration

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Liposomes were prepared by the thin-film hydration method.22 Briefly, SPC, Ch, CoQ10 and PTA were weighed at different ratios obtained from the experimental design and dissolved in 10 mL chloroform. The solution was transferred into a 250 mL round-bottomed flask, and the organic solvent was evaporated under a reduced pressure at 45°C and 100 rpm by a rotary evaporator (Heidolph, Schwabach, Germany). The resulting lipid film was dried overnight under a nitrogen gas flow to remove remaining traces of chloroform. The lipid film was hydrated with 10 mL phosphate-buffered saline (PBS; pH 7.4) by rotary evaporation at 45°C and 100 rpm without applying vacuum. For size reduction, liposomes were extruded 15 times through 200 nm pore-sized polycarbonate membranes (Whatman, Maidstone, UK) using the Avanti mini-extruder. The large unilamellar vesicles obtained were centrifuged at 3,000 g for 10 minutes in order to separate excess lipid and un-encapsulated drugs. Liposome formulations were maintained at 4°C and used within 3 days of preparation.
Çelik B., Sağıroğlu A.A, & Özdemir S. (2017). Design, optimization and characterization of coenzyme Q10- and D-panthenyl triacetate-loaded liposomes. International Journal of Nanomedicine, 12, 4869-4878.
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4

Preparation of Lipid Vesicles

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Desired amounts of lipids in chloroform were dried under a stream of nitrogen overnight under vacuum, followed by rehydration with appropriate buffer to 2–10 mg/mL concentration. For LUV preparation, the dispersion was frozen and thawed in liquid nitrogen and room temperature at least 5 times, followed by extrusion (15–21 times) through polycarbonate filters with 100 nm pores (Whatman) using Avanti Mini Extruder. For SUV preparation, the dispersion was sonicated with a tip sonicator (VibraCell VCX130) in 7–20 pulses lasting 10 s separated by 10 s breaks until the solution was clear.
Worch R., Dudek A., Borkowska P, & Setny P. (2021). Transient Excursions to Membrane Core as Determinants of Influenza Virus Fusion Peptide Activity. International Journal of Molecular Sciences, 22(10), 5301.
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5

Isolation and Characterization of Spinach Thylakoids

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Thylakoids were isolated from young spinach leaves using a modified method13 (link). The obtained thylakoids were pooled, diluted and sonicated for 2 min in a Fisher Scientific FS30D bath sonicator. This step was followed by extrusion through 100-nm polycarbonate porous membranes (Whatman) using an Avanti mini extruder. The solutions were then centrifuged for 60 min at 100,000g. The pellet was resuspended in osmotic shock buffer (10 mM HEPES-KOH, 10 mM MgCl2 and 10 mM sodium l-ascorbate). NanoSight NS300 (Malvern Instruments) was used to detect the concentration (particles per ml) of NTUs. The NTUs were flash-frozen with 10% DMSO as an osmoprotectant and stored at −80 °C until use. Before use, the NTUs were stored on ice and washed two to three times in osmotic shock buffer. A similar method was applied to encapsulate gold nanoparticles into the NTUs, and equal volumes of gold nanoparticles and thylakoids were mixed and then sonicated and extruded. The chlorophyll content of the resulting solution was determined using a chlorophyll assay kit (Acmec).
Chen P., Liu X., Gu C., Zhong P., Song N., Li M., Dai Z., Fang X., Liu Z., Zhang J., Tang R., Fan S, & Lin X. (2022). A plant-derived natural photosynthetic system for improving cell anabolism. Nature, 612(7940), 546-554.
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6

Preparation of Lipid Vesicles for Protein Studies

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A mixture of 2.75:1.25:1 mg of chloroform-dissolved 1-Palmitoyl-2-Oleoyl-sn-Glycero-3-Phosphoethanolamine (POPE): 1-Palmitoyl-2-Oleoyl-sn-Glycero-3-Phosphoglycerol (POPG):cholesterol (Anatrace) were dissolved and mixed in chloroform, later removed by evaporation. The resulting dried lipid film was then resuspended to 10 mg/ml in assay buffer, containing 150 mM NaCl, 1 mM EDTA, and 50 mM Na2HPO4-NaH2PO4, pH 6.0 (for experiments conducted with DOC2B, EDTA was excluded). The lipid mixture was then sonicated in a bath sonicator for 15 min, followed by extrusion through a 0.2 μm membrane (Whatman) for 11–13 times (Avanti mini-extruder), to form homogeneous, large unilamellar vesicles (LUVs), which were stored at 4 °C until use.
Manori B., Vaknin A., Vaňková P., Nitzan A., Zaidel-Bar R., Man P., Giladi M, & Haitin Y. (2024). Chloride intracellular channel (CLIC) proteins function as fusogens. Nature Communications, 15, 2085.
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