This experiment demonstrates the electro spinning of nanofiber scaffolds for extracellular structures of cell support in vitro and in vivo. Begin by choosing a polymer solvent and collector geometry suitable for the desired type of nano fibers. The most important step of this process is to determine the critical entanglement concentration above which it is possible to form fibers.
Next is to tailor the system to produce the diameter density and alignment of fibers. Desired electron microscopy indicates the different kinds of fibers that can be spun. These electro spun nanofiber scaffolds have been shown to accelerate the maturation, improve the growth, and direct the migration of cells in vitro ready.
Generally, individuals new to this method will struggle because electro spinning can be a finicky process. Visual demonstrations of this method is critical as the apparatus optimization steps are difficult to understand without any prior experience electro spinning. When choosing a polymer and solvent for electro spinning, consider characteristics like biodegradability, thermoplastic, and cross linkability with your eventual application in mind.
Then determine the suitable personal protective equipment required based on your selections. Next, select a substrate to collect the nano fibers on such as glass, plastic, metal, or silicon wafer. Now choose the collector geometry.
Randomly aligned fibers are collected on stationary plates. While aligned fibers can be collected on rapidly rotating wheels, drums, rods, or parallel plates ensure that the collector is conductive and remains isolated in such a way that it can be grounded without also grounding adjacent objects. In the case of rotating collectors, the wheel should be isolated from its axle.
In order to approximate the critical entanglement concentration empirically prepare several candidate polymer concentrations from these choose a concentration that flows. The solution should be a viscous liquid, but not a gel. Now set up the electro spinning apparatus.
The syringe should be placed at a 20 centimeter distance from the collector to start load the syringe pump with the chosen solution and set the pump speed such that any bead of solution wiped from the tip is immediately replaced. Ground the collector and clip the high voltage wire to the conductor plate. Make sure the power supply is set to zero before starting the wheel spinning while ramping up the voltage slowly watch the beat of the solution at the needle tip and observe the stream.
Continue to adjust the voltage to obtain a long and steady stream. If a steady stream cannot be obtained, adjust the polymer solution concentration first. If the stream is difficult to see, use a dark matte backdrop and place a unidirectional light source between the viewer and the stream.
At times, the polymer solution drips straight down from the syringe tip with no attraction to the wheel. In this case, confirm that the conductor plate is making contact with the needle tip and that the collector is making contact with ground. If the drop of polymer solution at the syringe tip is leaning in the direction of the wheel but is not forming a stream, increase the voltage.
Make adjustments by varying the distance and the voltage until a steady stream is visible. When large polymer globs form at the syringe tip, swipe the glob away with a paper towel attached to a non-conductive stick. In order to correct oscillating or wagging streams, turn down the voltage or increase the distance between the syringe tip and the wheel.
If the stream continues to wag, use a higher concentration of polymer or add a little solvent with a slower evaporation to the solution.Visible. Steady streams that make observable contact with the wheel set at to high rotational velocity yield the highest quality of uniformity and alignment to improve the length and steadiness of a stream that is short and discontinuous. Increase the polymer solution.
Add more slow evaporating solvent, or adjust the voltage. Sometimes when collecting random fibers on a stationary plate, fibers will begin to form sheets or yarns in midair. This is an indication that the voltage is set too high.
When beads are discovered in the fibers, increase the polymer solution and make sure the conductor plate is making continuous contact with the needle and that the grounded wire brush is making continuous contact with the wheel. Additionally, you should try to achieve a steady stream. Spitting streams often indicate the formation of beaded fibers.
If fibers form as ribbons or bleed together, use a higher concentration of polymer or a solvent with a higher rate of evaporation to correct fibers that form waves or curler cues. Increase the wheel speed or move the needle tip further from the collector. Also, check that the conductor plate and collector are not vibrating.
Sometimes pause a desired, in which case simply use a rapidly evaporating solvent. However, if pores are not desired, try adding a small amount of cos solvent that is less volatile than the major solvent. When the collector is moving at a low RPM or at rest, the alignment quality is poor.
Increase the alignment by increasing the speed of the wheel. Electros spun nano fibers are effective in the culture and study of various cell types. For example, primary rat neurons can be stained for beta tubulin and the nuclear counterstain dpi.
Importantly, the fibers can also be visualized when a dye such as sulfur rumine is included in the spinning solution After its development. This technique allowed researchers in the field of biomaterials and cell biology to explore the interactions between cells and a variety of nano topographical features and geometries. Don't forget that working with high voltages can be extremely hazardous, and precautions should always be taken to isolate the operator from the apparatus while performing this procedure.
