User:Karlena L. Brown/Notebook/PVOH Research/2013/02/13

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OBJECTIVES

  • Run fluorescence analysis on all hydrogel samples prepared with Rhodamine 6G dye (dye leaching out of medium anaylsis)
  • Grind microsphere PVOH 146K 110% CEC Laponite w/ DMHXLBR into a smaller sample
  • Re-evaluate strategy as to how to prepare more microspheres more efficiently

Fluorescence Analysis Instructions

  1. Start the FLWin Lab program
  2. Turn on the machine by clicking the on button on the side of the instrument
  3. Go to and select the Graph Button Command
  4. Afterwards select and the Setup Parameters Tab
  5. Within the Setup Parameters Tab, select the Emission Tab
  6. Set up parameters for fluorescence analysis by determining emission and excitation ranges
  7. Afterwards, turn on the lamp source by clicking on the Lamp Button in the instrument window
  8. Pour ~ 1-2mL of sample into a glass cuvette marked for fluorescence
  9. Then place the sample filled cuvette within the sample holder
  10. Next, in the Graph Window select the Lamp Button Icon to run the sample
  11. Once the sample has finished running, back in the main window select the Save As Command
  12. Finally officially save the sample measurement and file information as a text file under ASC II
  13. Create folder a to save the information under MK

Fluorescence Limit of Detection Calculations

Fluorescent Standards For Limit of Detection: 0.25μM, 0.5μM, 1μM, 92μM, & 165μM

0.25μM Rhodamine 6G Dye Concentration 

  M1V1 = M2V2
  0.25μM (RG6)x 5mL = (0.5μM)V2    V2 = 2.5mL

0.5μM Rhodamine 6G Dye Concentration 

  M1V1 = M2V2
  0.5μM (RG6)x 10mL = (92μM)V2    V2 = 54.34μL

1μM Rhodamine 6G Dye Concentration 

  M1V1 = M2V2
  1μM (RG6)x 25mL = (92μM)V2    V2 = 271.74μL

Notes

  • Microsphere sample PVOH 146K 110% CEC Laponite w/ DMHXLBR actually came out as a hunk of material rather than microspheres
  • For fluorescent analysis, several Rhodamine 6G dye standards were prepared in order to determine the limit of detection
  • Every sample previously prepared on 1/30/13 were analyzed because Rhodamine 6G dye was not crosslinked into hydrogel material
  • Specific parameters used to analyze fluorescence of hydrogel samples containing Rhodamine 6G dye included the following:
    # Excitation: 480nm
    # Emission Range: 500-650nm
    # Excitation Slit Width: 10
    # Emission Slit Width: 10
    # Scan Speed: 1200


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