User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/10/09: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> UV-vis Measurements & Sample Preparation</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> ZnPPIX Synthesis</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>}}
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==October 2, 2013==


==October 9, 2013==
===Procedures===
===Procedures===
*Absorbance measurements and Beer's law calculations for samples from 10/25/2013
*Synthesize new ZnPPIX with Dr Hartings
*Sample preparation based on calculated concentrations
*Procedure adapted from "On the preparation of metalloporphyrins"
**0.5g tetraphenylporphyrin refluxed in 50mL DMF
**0.403g (stiochiometric amount) zinc acetate added, solution refluxed until reaction complete
***completion determination by unchanged peaks in UV-vis measurements
**50mL distilled water added, solution chilled and then filtered
***the filtrate was filtered over again and added to main filtered sample


===Data===
===Data===
=====Absorbance Measurements from 10/25/2013=====
Absorbance Measurements
[[Image:2013_1002_ZnPPIX_from_last_week.PNG]]<br.>
*Used to determine reaction completion (based on the change in the peaks)
Beer's Law calculations:<br.>
[[Image:2012 1009 ZnPPPIX abs.PNG]]
[[Image:2013_1002_ZnPPIX_from_last_week_calcs.PNG]]
No visible change in peaks between 'ZnPPIX#2 100x' (dark blue line- the second sample of refluxing ZnPPIX taken) and 'ZnPPIX after' (light blue line- the final sample taken when reflux was stopped), used to determine that the reaction was complete.<br.>
 
Peaks consistent with the literature: largest peak at ~500nm, next largest at ~600nm, smallest peak at ~550nm.
=====Absorbance Measurements, Today=====
I decided to run two more samples at higher concentrations on the UV-vis and calculate the concentrations from these because I needed higher concentrations in order to make my samples at the concentrations I've been using.
<br.>
[[Image:2013_1002_ZnPPIX_for_calcs.PNG]]<br.>
Beer's Law calculations:<br.>
[[Image:2013_1002_ZnPPIX_calcs.PNG]]
 
=====Samples Prepared=====
*2.47uM ZnPPIX
*2.47uM ZnPPIX/1.11uM DNA-SH
*2.47uM ZnPPIX/1.11uM DNA (4)
*2.47uM ZnPPIX/1.11uM DNA (8)
*1.934uM ZnPPIX/0.94uM DNA-SH/11.75nM AuNPs
*1.934uM ZnPPIX/0.74uM DNA (4)/11.75nM AuNPs
*1.934uM ZnPPIX/0.74uM DNA (8)/11.75nM AuNPs
*2.47uM ZnPPIX/11.75nM AuNPs


===Notes===
===Notes===
*Run today's samples for absorbance and fluorescence
Next time:
*Ask Dr Hartings for more ZnPPIX
*Dissolve filtered ZnPPIX in DMF
 
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Revision as of 08:31, 16 October 2013

ZnPPIX Synthesis <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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October 9, 2013

Procedures

  • Synthesize new ZnPPIX with Dr Hartings
  • Procedure adapted from "On the preparation of metalloporphyrins"
    • 0.5g tetraphenylporphyrin refluxed in 50mL DMF
    • 0.403g (stiochiometric amount) zinc acetate added, solution refluxed until reaction complete
      • completion determination by unchanged peaks in UV-vis measurements
    • 50mL distilled water added, solution chilled and then filtered
      • the filtrate was filtered over again and added to main filtered sample

Data

Absorbance Measurements

  • Used to determine reaction completion (based on the change in the peaks)

No visible change in peaks between 'ZnPPIX#2 100x' (dark blue line- the second sample of refluxing ZnPPIX taken) and 'ZnPPIX after' (light blue line- the final sample taken when reflux was stopped), used to determine that the reaction was complete.<br.> Peaks consistent with the literature: largest peak at ~500nm, next largest at ~600nm, smallest peak at ~550nm.

Notes

Next time:

  • Dissolve filtered ZnPPIX in DMF


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