User:Nancy T. Miles/Notebook/CHEM 572 Spr 2014/2014/04/01

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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|style="background-color: #EEE"|[[Image:BDLlogo_notext_lr.png|128px]]<span style="font-size:22px;"> Biomaterials Design Lab</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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==Entry title==
+
 
-
* Insert content here...
+
==Objectives===
 +
*Making new scaffolds
 +
*New AgNP procedures
 +
*Prepare samples for bacteria culture
 +
 
 +
==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 [http://pubs.acs.org/doi/pdf/10.1021/ed084p322 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
 +
 

Current revision

Biomaterials Design Lab Main project page
Previous entry      Next entry

Objectives=

  • Making new scaffolds
  • New AgNP procedures
  • Prepare samples for bacteria culture

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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