User:Allison K. Alix/Notebook/CHEM-581/2013/04/03

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Objective

-Mix hydrogels with remaining indicators (bromocresol purple, phenolphthalein, cresol red)

-View hydrogels containing fluorescent dye (R6G) under fluorescent microscope

-Prepare new sets of hydrogels

Procedures

Part 1: Addition of indicators to hydrogel solutions

1) Add 50μL of phenolphthalein to 5mg hydrogel in 1.5mL buffer prepared on 03/29/2013

2) Add 25μL bromocresol purple to 5mg hydrogels in 1.5mL phosphate buffer

3) Add 75μL 0.02% cresol red to 5mg hydrogels in 1.5mL phosphate buffer

4) Allow time for hydrogels to absorb indicator

Part 2: Create concentration gradient for fluroescence calibration of microscope

1) Prepare a solution of 10μL FluoSpheres carboxylate modified microspheres, (0.1μM, orange fluorescent (540/560) 2% solids) in 100μL distilled water (solution 1) 0.2%

2) Prepare the following subsequent dilutions

  • solution 2: 10μL solution 1 in 100μL distilled water 0.02%
  • solution 3: 10μL solution 2 in 100μL distilled water 0.002%
  • solution 4: 10μL solution 3 in 100μL distilled water 0.0002%

Part 3: Preparing dilutions of dye containing hydrogels (marked with *)

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

  • Dilute 200μL of solution 1 with 200μL phosphate buffer (solution 2)
  • Dilute 200μL of solution 2 with 200μL phosphate buffer (solution 3)
  • Dilute 200μL of solution 3 with 200μL phosphate buffer (solution 4)
  • Dilute 200μL of solution 4 with 200μL phosphate buffer (solution 5)

2) 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 200μL of solution 1 with 200μL phosphate buffer (solution 2)
  • Dilute 200μL of solution 2 with 200μL phosphate buffer (solution 3)
  • Dilute 200μL of solution 3 with 200μL phosphate buffer (solution 4)
  • Dilute 200μL of solution 4 with 200μL phosphate buffer (solution 5)

Part 4: Preparation of hydrogels

1) Prepare the following:

  • 2 samples of 0.5g (89,000-98,000 g/mol) PVOH in 6mL phosphate buffer (labelled A and B)
  • 2 samples of 0.5g (89,000-98,000 g/mol) PVOH in 6mL phosphate buffer with 300μL 1mM R6G (labelled C and D)

2) Heat solutions at ~110°C for 15 minutes. Allow to cool to room temperature.

3) mix with 40mL safflower oil for 1-2min on lower setting of blender

4) Freeze with liquid nitrogen and place in freezer at -20°C for 24 hours

Data

  • Add data and results here...