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Figure 1.Fiber analysis: electrospun nitrocellulose. SEM images reveal that nitrocellulose forms fibers over a narrow range of electrospinning conditions. Samples electrospun from less than 110 mg/ml underwent electrospraying, a process that occurs when polymer chain entanglements are inadequate to induce fiber formation. Fibers were evident in samples prepared from 110 mg/ml, 120 mg/ml and 140 mg/ml. Note nearly uniform fiber diameters in 110 mg/ml samples, heterogeneity of diameters present in 120 mg/ml samples and solvent defects in 140 mg/ml samples (upper left of image). Fibers in 110 mg/ml solutions were smaller than fibers produced from the 120 and 140 mg/ml starting concentrations (P μm.
Figure 2.Viscosity as a function of starting concentration. (A). Viscosity was similar in solutions prepared with 60, 80 and 100 mg/ml nitrocellulose. Regression analysis of the entire data set using a 1st order equation generated an R2 value of 0.677, applying a 1st order equation to the range of starting concentrations that produced fibers (100–140 mg/ml) generated an R2 of 0.978. (B). Viscosity increased markedly from 100 to 140 mg/ml (panel A), however there was no clear relationships between fiber diameter and viscosity (panel B).
Figure 3.Representative Slot Blots. Chemiluminescence detection of Fn on control nitrocellulose (Lane A), corresponding image of oxidized Lumigen reaction product (B). Chemiluminescence detection of Fn on electrospun nitrocellulose (Lane C), corresponding image of oxidized Lumigen reaction product (D). Graphical illustration depicting the relative optical density present in slot blots (E). Conventional nitrocellulose blot exhibited modest increase in chemiluminescence signal as a function of increasing Fn concentration (R2 = 0.655). Electrospun nitrocellulose exhibited a more pronounced signal at all protein concentrations examined. Signal increased in a nearly linear fashion over a broad range of concentrations (R2 = 0.905).
Figure 4.Fiber analysis: electrospun nylon. SEM images indicated that fibers were produced under all conditions assayed. Fibers electrospun from the 60 mg/ml solutions were smaller than all other treatment groups (P
Figure 5.Viscosity as a function of starting concentration. (A). Solution viscosity for nylon prepared in HFIP increased as a 1st order function (R2 = 0.809), a 2nd order equation provided an R2 = 0.989. Fiber diameter did not appear to be directly related to viscosity of the starting solutions, fiber diameter remained nearly constant over a wide range of starting conditions and solution viscosities (B).
Figure 6. Electrospun membranes in Western blotting applications. (A). Chemiluminescence detection of Fn (lanes 1 to 9 = 10.0, 5.0, 2.5, 1.0, 0.50, 0.25, 0.16 0.08, 0.02 μg Fn/lane) on parent nitrocellulose and electrospun nitrocellulose, parent nylon and electrospun nylon (using 1 μm diameter fibers). Electrospun nitrocellulose exhibited high signal, but poor band resolution. The parent nylon material exhibited a diffuse signal in lanes loaded with the highest concentration of Fn and prominent negative images (A, lanes 1–5, top right). Contrast with electrospun nylon, this membrane exhibited sharper bands and no evidence of inverse image formation. (B) Electrospun nylon/electrospun nitrocellulose fiber composite. Note: high signal, absence of inverse white bands, but poor band resolution. We associate this result with a composite that is too thick. SEM images before (Bar = 10 μm) and after (Bar = 5 μm) blocking buffers applied to composite, inset demonstrating material coating fibers, pores between adjacent fibers remain present and open. These images suggested that electrospun nitrocellulose fibers underwent an increase in diameter during blotting (C). Electrospun nylon/electrospray nitrocellulose bead composite. This composite provided good signal detection and band resolution. SEM images before (Bar = 10 μm) and after (Bar = 5 μm) blocking buffers applied to composite.
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