User:Samiye Yaman/Notebook/Lab 581/2013/04/03: Difference between revisions

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==Description==
==Description==
 
'''Part A: pH Analysis'''
 
*Add 50μL of phenolphthalein to 5mg hydrogel in 1.5mL buffer prepared on 03/29/2013.
 
*Add 25μL bromocresol purple to 5mg hydrogels in 1.5mL phosphate buffer.
 
* Add 75μL 0.02% cresol red to 5mg hydrogels in 1.5mL phosphate buffer. Allow some time for hydrogels to absorb indicators.
 
'''Part B: Create concentration gradient for fluroescence calibration of microscope'''
 
* Add 10μL FluoSpheres carboxylate modified microspheres in to 100μL distilled water. This is solution one. Take 10μL from solution 1 and place in to a seperate falcon tube. Add 100μL distilled water. This is solution 2. Solution 3 is created by mixing 10μL of solution from solution 1 and 100μL distilled water. Repeat the same process for creating solution 4 and 5. (Diluting each solution by 10)
 
'''Part C: Preparing dilutions of dye containing hydrogels'''
 
* Dilute 2mg hydrogels containing 30μL R6G (supernatant was previously removed) with 400μL phosphate buffer (solution 1).
 
-Dilute each solution by half. (take 200μL solution from solution 1 and mix it with 200 phosphate buffer to create solution 3.)
 
* Dilute 200μL 2mg of hydrogel solution containing dye in 1.5mL phosphate buffer (from 3/22/13) (marked with heart) with 200μL phosphate buffer (solution 1)
 
- Dilute each solution by half. (take 200μL solution from solution 1 and mix it with 200 phosphate buffer to create solution 3.)
 
'''Part D: Synthesizing more hydrogels'''
 
* Add 0.5g (89,000-98,000 g/mol) PVOH in 6mL phosphate buffer into 2 beakers labelled as A and B.
 
* Add 0.5g (89,000-98,000 g/mol) PVOH in 6mL phosphate buffer with 300μL 1mM R6G into 2 beaker labelled as C and D.
 
* Place the beakers on a heat block at 110°C for 15 minutes.
 
*Let them cool to room temperature.
 
*By using blender, mix each hydrogels with 40mL safflower oil for 1-2min on lower setting of blender.
 
*Shock freeze the blended hydrogels with oil by using liquid nitrogen and place in freezer at -20°C for 24 hours.
 
*After in freezer for 24 hours, take them into room temperature for 4 hours.(cycle 1). The cycles are required.
 
==Data==
==Data==


==Notes==
==Notes==

Revision as of 15:21, 6 April 2013

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Objections

Description

Part A: pH Analysis

  • Add 50μL of phenolphthalein to 5mg hydrogel in 1.5mL buffer prepared on 03/29/2013.
  • Add 25μL bromocresol purple to 5mg hydrogels in 1.5mL phosphate buffer.
  • Add 75μL 0.02% cresol red to 5mg hydrogels in 1.5mL phosphate buffer. Allow some time for hydrogels to absorb indicators.

Part B: Create concentration gradient for fluroescence calibration of microscope

  • Add 10μL FluoSpheres carboxylate modified microspheres in to 100μL distilled water. This is solution one. Take 10μL from solution 1 and place in to a seperate falcon tube. Add 100μL distilled water. This is solution 2. Solution 3 is created by mixing 10μL of solution from solution 1 and 100μL distilled water. Repeat the same process for creating solution 4 and 5. (Diluting each solution by 10)

Part C: Preparing dilutions of dye containing hydrogels

  • Dilute 2mg hydrogels containing 30μL R6G (supernatant was previously removed) with 400μL phosphate buffer (solution 1).

-Dilute each solution by half. (take 200μL solution from solution 1 and mix it with 200 phosphate buffer to create solution 3.)

  • Dilute 200μL 2mg of hydrogel solution containing dye in 1.5mL phosphate buffer (from 3/22/13) (marked with heart) with 200μL phosphate buffer (solution 1)

- Dilute each solution by half. (take 200μL solution from solution 1 and mix it with 200 phosphate buffer to create solution 3.)

Part D: Synthesizing more hydrogels

  • Add 0.5g (89,000-98,000 g/mol) PVOH in 6mL phosphate buffer into 2 beakers labelled as A and B.
  • Add 0.5g (89,000-98,000 g/mol) PVOH in 6mL phosphate buffer with 300μL 1mM R6G into 2 beaker labelled as C and D.
  • Place the beakers on a heat block at 110°C for 15 minutes.
  • Let them cool to room temperature.
  • By using blender, mix each hydrogels with 40mL safflower oil for 1-2min on lower setting of blender.
  • Shock freeze the blended hydrogels with oil by using liquid nitrogen and place in freezer at -20°C for 24 hours.
  • After in freezer for 24 hours, take them into room temperature for 4 hours.(cycle 1). The cycles are required.

Data

Notes
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