User:Alicia Rasines Mazo/Notebook/CHEM-581 Experimental Chemistry I/2014/10/01: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</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"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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## 0.10μM
## 0.10μM
## 0.50μM
## 0.50μM
## 0.75μM
## 1.0μM
## 1.0μM
## 1.2μM
## 1.5μM
## 1.5μM
## 2.0μM
## 2.0μM
#* Due to extremely low amounts of R6G required to make stock solutions, an initial stock solution of 2500 μM was prepared, this solution was further diluted to 500μM to make rest of solutions, except for 2.0 μM, which was prepared using 2500μM stock solution
#* Due to extremely low amounts of R6G required to make stock solutions, an initial stock solution of 2500 μM was prepared. This solution was further diluted to 500μM to make rest of solutions, except for 2.0 μM, which was prepared using 2500μM stock solution
#**(2500×10<sup>-6</sup>mol/L)*(0.010L)*(474.02g/mol)=0.01185 g ('''0.0120 g of R6G were measured out)
#**(2500×10<sup>-6</sup>mol/L)*(0.010L)*(474.02g/mol)=0.01185 g ('''0.0120 g of R6G were measured out)
#*** 2μM stock concentration: (2500μM)v<sub>1</sub>=(2.0μM)(5mL);  '''v<sub>1</sub>'''=0.004mL
#*** 2μM stock concentration: (2500μM)v<sub>1</sub>=(2.0μM)(5mL);  '''v<sub>1</sub>'''=0.004mL
#** 500μM dilution: (2500μM)v<sub>1</sub>=(500μM)(5mL);  '''v<sub>1</sub>'''=1mL of 2500μM R6G stock solution
#** 500μM dilution: (2500μM)v<sub>1</sub>=(500μM)(5mL);  '''v<sub>1</sub>'''=1mL of 2500μM R6G stock solution
#*** 1.5μM stock solution: (500μM)v<sub>1</sub>=(1.5μM)(5mL);  '''v<sub>1</sub>'''=0.0015mL of 500μM stock solution (Same calculations for rest of stock solutions)
#*** 1.5μM stock solution: (500μM)v<sub>1</sub>=(1.5μM)(5mL);  '''v<sub>1</sub>'''=0.0015mL of 500μM stock solution (Same calculations for rest of stock solutions)
#*** 1.0μM stock solution: 0.010mL of 500μM stock solution were diluted to 5mL with distilled water
#*** 1.2μM stock solution: 0.012mL of 500μM stock solution were diluted to a total volume of 5mL with distilled water
#*** 0.50μM stock solution: 0.005mL of 500μM stock solution were diluted to 5mL with distilled water
#*** 1.0μM stock solution: 0.010mL of 500μM stock solution were diluted to a total volume of 5mL with distilled water
#*** 0.1μM stock solution: 0.001mL of 500μM stock solution were diluted to 5mL with distilled water
#*** 0.75μM stock solution: 0.0075mL of 500μM stock solution were diluted to a total volume of 5mL with distilled water
#*** 0.50μM stock solution: 0.005mL of 500μM stock solution were diluted to a total volume of 5mL with distilled water
#*** 0.1μM stock solution: 0.001mL of 500μM stock solution were diluted to a total volume of5mL with distilled water
# Recorded UV-Vis and Fluorescence spectra of these samples.
# Recorded UV-Vis and Fluorescence spectra of these samples.
#* Excitation: 530nm
#* Excitation: 500nm
#* Excitation range: 545 to 700 nm
#* Excitation range: 515 to 700 nm
#* Slit width: 10 nm
#* Slit width: 10 nm
#* Scan speed: 200 nm/min
#* Scan speed: 200 nm/min
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# Make a calibration curve based on the fluorescence.
# Make a calibration curve based on the fluorescence.
## In order to do this, you'll need to measure the area under the fluorescence curve, not just the fluorescence peak height.
## In order to do this, you'll need to measure the area under the fluorescence curve, not just the fluorescence peak height.
===Calibration curve of R6G using UV-Vis===
[[Image:R6G UVVIS calibration.png]]<br.>
[[Image:MolarAbsorp R6G 521.png]]
===Calibration curve of R6G based on fluorescence===
[[Image:Fluorescence R6G.png]]<br.>
[[Image:Fluorescence R6G calibrationcurve.png]]
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__NOTOC__
__NOTOC__

Latest revision as of 00:21, 27 September 2017

Project name Main project page
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Task for October 1

  • To determine fluorophore concentration

Fluorescence

Procedure followed as directed by Dr. Hartings on Oct. 1<br.>

  1. Made stock concentrations of Rhodamine 6G (both us and Eleni's group used the same solutions)
    1. 0.10μM
    2. 0.50μM
    3. 0.75μM
    4. 1.0μM
    5. 1.2μM
    6. 1.5μM
    7. 2.0μM
    • Due to extremely low amounts of R6G required to make stock solutions, an initial stock solution of 2500 μM was prepared. This solution was further diluted to 500μM to make rest of solutions, except for 2.0 μM, which was prepared using 2500μM stock solution
      • (2500×10-6mol/L)*(0.010L)*(474.02g/mol)=0.01185 g (0.0120 g of R6G were measured out)
        • 2μM stock concentration: (2500μM)v1=(2.0μM)(5mL); v1=0.004mL
      • 500μM dilution: (2500μM)v1=(500μM)(5mL); v1=1mL of 2500μM R6G stock solution
        • 1.5μM stock solution: (500μM)v1=(1.5μM)(5mL); v1=0.0015mL of 500μM stock solution (Same calculations for rest of stock solutions)
        • 1.2μM stock solution: 0.012mL of 500μM stock solution were diluted to a total volume of 5mL with distilled water
        • 1.0μM stock solution: 0.010mL of 500μM stock solution were diluted to a total volume of 5mL with distilled water
        • 0.75μM stock solution: 0.0075mL of 500μM stock solution were diluted to a total volume of 5mL with distilled water
        • 0.50μM stock solution: 0.005mL of 500μM stock solution were diluted to a total volume of 5mL with distilled water
        • 0.1μM stock solution: 0.001mL of 500μM stock solution were diluted to a total volume of5mL with distilled water
  2. Recorded UV-Vis and Fluorescence spectra of these samples.
    • Excitation: 500nm
    • Excitation range: 515 to 700 nm
    • Slit width: 10 nm
    • Scan speed: 200 nm/min
  3. Make a calibration curve based on UV-Vis.
    1. Compare your data to some published values
  4. Make a calibration curve based on the fluorescence.
    1. In order to do this, you'll need to measure the area under the fluorescence curve, not just the fluorescence peak height.

Calibration curve of R6G using UV-Vis

<br.>

Calibration curve of R6G based on fluorescence

<br.>


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