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.