User:Madeleine Y. Bee/Notebook/CHEM-572 2014S/2014/04/01

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Scaffold Prep, and AgNPs without BSA Main project page
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April 1, 2014

Objectives

  • Making new scaffolds
  • New AgNP procedures

Au:Myoglobin Nanofibers: Sample Prep for Cell Culture

Image:2014_0211_myo_bsa_NPs.PNG

  • Solutions:
    • TRIS dopamine: 0.0475g dopamine hydrochloride and 0.03g TRIS in 25mL to make 10mM solution at pH 8.46
    • Au: 0.011g in 10mL to make 2.793mM solution
    • Myoglobin: 0.011g in 10mL to make 0.0621mM solution

Plain Scaffolds

  • 4 PLA scaffolds printed for each test tube
  • Three test tubes filled with scaffolds and water
  • Autoclaved

Brute Force Nanofiber-coated Scaffolds

  • 4 PLA scaffolds printed for each test tube
  • Gold:Myoglobin (100:1) nanofibers synthesized by above method
  • Nanofibers dried onto PLA scaffolds
  • Scaffolds submerged in water
  • Autoclaved

Original Method for Nanofiber-coated Scaffolds

  • 4 PLA scaffolds printed for each test tube
  • Gold:Myoglobin (100:1) nanofibers synthesized by above procedure with scaffolds in solution
  • Autoclaved

Polydopamine-coated Scaffolds

  • 4 PLA scaffolds printed for each test tube
  • Scaffolds submerged in 10mM pH 8.46 TRIS dopamine hydrochloride solution for 1 hour
  • Add 1mL of 0.0621 Myoglobin to each test tube, shake tube to combine, allow surface functionalization to occur for 5 hours
  • Rinse with deionized water and allow scaffold to dry overnight
  • Scaffolds submerged in water
  • Autoclaved

Polydopamine Nanofiber-coated Scaffolds

  • 4 PLA scaffolds printed for each test tube
  • Scaffolds submerged in 10mM pH 8.5 TRIS dopamine hydrochloride solution for 1 hour
  • Add 1mL of 0.0621 Myoglobin to each test tube, shake tube to combine, allow surface functionalization to occur for 5 hours
  • Rinse with deionized water and allow scaffold to dry overnight
  • Gold:Myoglobin (100:1) nanofibers synthesized by above procedure with polydopamine-coated scaffolds in solution
  • Autoclaved

Breakdown

Today
  • Time sensitive: Make 40mL TRIS-polydopamine solution
  • Time sensitive: Make 6 tests tubes of TRIS-polydopamine solution, put scaffolds in all 6 test tubes at 12 p.m. (3 for pD only, 3 for pD-original)
  • Make Au and Myoglobin solutions
  • Make 9 test tubes of 100:1 Au:Myo Nanofibers (3 for original, 3 for brute force fibers, 3 for pD-original tomorrow; see volumes in chart above)
  • Time sensitive: At 1 p.m. add 1mL of 0.0621M Myoglobin to all 6 tubes (3 for pD only, 3 for pD-original)
  • Put scaffolds into 3 of the 9 Au:Myo test tubes when finished (3 for original)
  • Make 3 test tubes of water -> put scaffolds into 3 test tubes when finished
  • Put 9 test tubes (3 brute force, 3 original, 3 water) into oven, set for 4 hours at 80C
  • Time Sensitive: At 6 p.m. rinse pD scaffolds that have soaked for 5 hours, submerge 6 in water overnight (if possible, submerge 3 of 6 tubes in Au:Myo solution and put all 6 in oven for 4 hours), dump Au:Myo fibers onto scaffolds for brute force
Tomorrow
  • If oven was not finished yesterday: submerge 3 pD scaffolds into original solution, leave other 3 in water and put in oven for 4 hours at 80C
  • Submerged dried brute force scaffolds in water
  • Autoclave all: 3 plain, 3 brute force, 3 original, 3 pD-Myo, 3 pD-Myo-Au

Sample Preparation: AgNPs

Without BSA
  • From Synthesis and Study of Silver Nanoparticles
  • 10 ml of 1 mM AgNO3 was added dropwise (about 1 drop/sec) to 30 ml of 2 mM NaBH4 (cooled in ice bath), while the reaction mixture was vigorously stirred using a magnetic stir plate. Entire procedure takes about 3 minutes.

Sample Preparation: Bacteria Culture

  • Make solution
  • Autoclave



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