User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/07/08

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Current revision (08:55, 10 July 2013) (view source)
(FCS Measurements)
 
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Meeting, FCS Measurements</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"|<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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==July 8, 2013==
==July 8, 2013==
===Notes from Meeting===
===Notes from Meeting===
-
*
+
*Since we're seeing some noise from just PBS on the FCS: make new PBS and run, parse out 10mL samples to avoid contaimination
 +
*Sonicate or centrifuge green fluorescent beads before running to prevent aggregation while measurements are being taken
 +
*Go up to 500pM for Oligo D samples (and lower concentrations)
 +
*Use a high, constant concentration of DNA with lower, decreasing concentration of MB
 +
*Check for consistent diffusion times between samples run on different days
 +
**Different diffusion times indicate background is something other than unbound DNA
 +
*Look up AuNP/DNA/MB fluorescent data and run samples/make and run new samples
 +
*Look in literature for ways to calculate silica shell thickness without TEM results
 +
===FCS Sample Prep===
 +
*Green fluorescent beads
 +
**10000x diluted
 +
**Run 3x, 90s
 +
*Oligo D
 +
**150pM
 +
**125pM
 +
**100pM
 +
**Run 3x, 500s
 +
*DNA/MB: DNA held at a constant concentration to isolate background from DNA
 +
**600/500
 +
**600/400
 +
**600/300
 +
**600/200
 +
**600/100
 +
**Run 3x, 500s

Current revision

Meeting, FCS Measurements Main project page
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July 8, 2013

Notes from Meeting

  • Since we're seeing some noise from just PBS on the FCS: make new PBS and run, parse out 10mL samples to avoid contaimination
  • Sonicate or centrifuge green fluorescent beads before running to prevent aggregation while measurements are being taken
  • Go up to 500pM for Oligo D samples (and lower concentrations)
  • Use a high, constant concentration of DNA with lower, decreasing concentration of MB
  • Check for consistent diffusion times between samples run on different days
    • Different diffusion times indicate background is something other than unbound DNA
  • Look up AuNP/DNA/MB fluorescent data and run samples/make and run new samples
  • Look in literature for ways to calculate silica shell thickness without TEM results

FCS Sample Prep

  • Green fluorescent beads
    • 10000x diluted
    • Run 3x, 90s
  • Oligo D
    • 150pM
    • 125pM
    • 100pM
    • Run 3x, 500s
  • DNA/MB: DNA held at a constant concentration to isolate background from DNA
    • 600/500
    • 600/400
    • 600/300
    • 600/200
    • 600/100
    • Run 3x, 500s



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