Unidirectionally aligned and randomly distributed nanofibers were fabricated from poly(L-lactic acid) (PLLA, Mw = 152,000, Sigma-Aldrich) by electrospinning. Polymer solution was prepared by dissolving PLLA in dichloromethane (DCM)/n,n-dimethylformamide (DMF) (70/30) at 4% w/w. The solution was loaded into a 1 ml syringe with a 30-gauge needle and dispensed at 1 ml/h with a syringe pump. A voltage of 18 kV was applied using a DC power supply to generate the polymer jet. The electrospinning distance was fixed at 15 cm. For producing aligned nanofibers, a rotating collection disk was used (rotating at 1000 rpm). For random nanofibers, a flat stationary plate was used for collection. Both aligned and random nanofibers were collected on spin-cast PLLA films formed on 18 mm dia. round glass coverslips. Spin-casting of PLLA films was conducted at 4000 rpm for 25 sec using a 1% w/w PLLA solution in chloroform. Spin-cast PLLA films were also used as flat controls in cell culture assays. For scanning electron microscopy (SEM), nanofibers were coated with platinum/palladium using the sputter coater (Ted Pella) and observed by Quanta 200F SEM (FEI). Nanofiber diameter was measured from SEM images with ImageJ software.
Sputter coater
Manufactured by Ted Pella
The Sputter Coater is a versatile laboratory equipment used to apply thin, uniform coatings of conductive materials onto samples in preparation for various analytical techniques. The core function of the Sputter Coater is to create a vacuum environment and use a high-voltage electric field to eject atoms from a target material, which then condense onto the sample surface, forming a thin film.
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4 protocols using sputter coater
1
Electrospun PLLA Nanofiber Fabrication
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Characterizing Hierarchical Porous Materials
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Transmission electron microscopy (TEM, Tecnai F30, FEI) and scanning electron microscopy (SEM, Carl Zeiss, Merlin) were used to characterize the hierarchical porous materials and material/cell interfaces. CMs on the materials were fixed in 5% glutaraldehyde PBS solution for 30 min, washed in DI water, and then dehydrated with an increasing ethanol gradient from 30% to 98%. The samples were dried in a critical point dryer (Leica EM CPD300) and observed on the same SEM after coating with an 8 nm Pt/Pd metal layer on a sputter coater (Ted Pella, Inc.). The SEM was operated at a 2-kV accelerating voltage. Images were analysed using ImageJ.
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Characterization of Arsenic Sulfide Nanomaterials
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Arsenic sulfide precipitate was separated from cells and media by centrifugation in 50 ml conical tubes. Precipitates were washed and resuspended, five times in DDI water to remove salts. After the washing process, the precipitate was resuspended in a 1 ml DDI water and deposited on a silicon wafer (Ted Pella) and air dried overnight. Samples were coated with gold in a sputter coater (Ted Pella), and scanning electron micrographs were obtained using JEOL 700 FE scanning electron microscopy, accessorized with energy‐dispersive X‐ray spectroscopy (EDX) for elemental analysis. Crystallinity of the nanomaterials was analysed by depositing the DI water washed nanomaterials on a glass slide and using a X‐ray diffraction (XRD) scans on a Rigaku Ultima IV diffractometer.
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Characterizing Hierarchical Porous Materials
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