User:Madeleine Y. Bee/Notebook/CHEM-572 2014S/2014/04/02: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> </span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Nanofiber-coated scaffolds and Ag:BSA fibers</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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*Finish up Au:Myo Nanofiber scaffolds for cell culture
*Finish up Au:Myo Nanofiber scaffolds for cell culture
*Make AgNPs with BSA according to article
*Make AgNPs with BSA according to article
*
*UV-vis of finished NPs


===Au:Myoglobin Nanofibers: Sample Prep for Cell Culture===
===Au:Myoglobin Nanofibers: Sample Prep for Cell Culture===
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*From [http://www.sciencedirect.com/science/article/pii/S0021979712009526 Characterization of Silver/Bovine Serum Albumin (Ag/BSA) nanoparticles structure: Morphological, compositional, and interaction studies]
*From [http://www.sciencedirect.com/science/article/pii/S0021979712009526 Characterization of Silver/Bovine Serum Albumin (Ag/BSA) nanoparticles structure: Morphological, compositional, and interaction studies]
*"Initially 0.9 g of BSA was mixed with 40 ml of deionized water. One milli liter of 0.423 M silver nitrate was added to BSA solution at ∼23 °C and sonicated for 10 min. This was followed by the addition of 1 ml of 0.423 M of sodium borohydride and sonicated for additional 30 min to reduce the silver ions in the solution. The resulting brown homogeneous solution was centrifuged at 21,500 rpm and the residue was washed with ethanol, suspended in water, and saved in an amber colored bottle at 4 °C."
*"Initially 0.9 g of BSA was mixed with 40 ml of deionized water. One milli liter of 0.423 M silver nitrate was added to BSA solution at ∼23 °C and sonicated for 10 min. This was followed by the addition of 1 ml of 0.423 M of sodium borohydride and sonicated for additional 30 min to reduce the silver ions in the solution. The resulting brown homogeneous solution was centrifuged at 21,500 rpm and the residue was washed with ethanol, suspended in water, and saved in an amber colored bottle at 4 °C."
*Procedure from yesterday: 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.
**Dilute solutions from first procedure to redo procedure from yesterday, combining BSA and silver nitrate before adding then drop-wise to the cooled, stirred sodium borohydride






*Cell tests
===Type I Collagen Test (Detection and Quantification)===
*[http://www.chondrex.com/products/mouse-type-i-collagen-detection-kit Company procedure ]
===Collagen Staining Procedure & Microscope Imaging===
*[http://www.chondrex.com/products/sirius-red-fast-green-collagen-staining-kit Company procedure]
===Trypsin Cell Detachment Procedure & Cell Counting===
*[http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/General_Information/1/celldetachmentprotocol.pdf Company procedure]


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Revision as of 17:40, 18 April 2014

Nanofiber-coated scaffolds and Ag:BSA fibers <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

April 2, 2014

Objectives

  • Finish up Au:Myo Nanofiber scaffolds for cell culture
  • Make AgNPs with BSA according to article
  • UV-vis of finished NPs

Au:Myoglobin Nanofibers: Sample Prep for Cell Culture

  • Continued from yesterday
Finishing Up
  • 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

  • Continued from yesterday
With BSA (direct from article)
  • From Characterization of Silver/Bovine Serum Albumin (Ag/BSA) nanoparticles structure: Morphological, compositional, and interaction studies
  • "Initially 0.9 g of BSA was mixed with 40 ml of deionized water. One milli liter of 0.423 M silver nitrate was added to BSA solution at ∼23 °C and sonicated for 10 min. This was followed by the addition of 1 ml of 0.423 M of sodium borohydride and sonicated for additional 30 min to reduce the silver ions in the solution. The resulting brown homogeneous solution was centrifuged at 21,500 rpm and the residue was washed with ethanol, suspended in water, and saved in an amber colored bottle at 4 °C."
  • Procedure from yesterday: 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.
    • Dilute solutions from first procedure to redo procedure from yesterday, combining BSA and silver nitrate before adding then drop-wise to the cooled, stirred sodium borohydride




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