User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/07/05

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(July 5, 2013)
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===FCS Sample Prep===
===FCS Sample Prep===
*Green fluorescent beads
*Green fluorescent beads
-
**10000x diluted
+
**10000x diluted, run 3x for 90s
 +
*PBS
 +
**To check for background, run 3x for 200s
*DNA/MB
*DNA/MB
-
**125/100pM
+
**125/100pM, run 3x for 500s
-
**100/80pM
+
**100/80pM, run 3x for 500s
-
**75/60pM
+
**75/60pM, run 3x for 500s
-
**50/40pM
+
**50/40pM, run 3x for 500s
-
**25/20pM
+
**25/20pM, run 3x for 500s
===FCS Data===
===FCS Data===
-
====Notes====
+
Graphs generated using Igor Pro.<br.>
 +
<br.>
 +
[[Image:Avg_Beads.png]]<br.>
 +
10000x diluted green fluorescent beads were used to align the laser.  Sample was run three times, for 90s each.<br.>
 +
<br.>
 +
[[Image:Avg_PBS.png]]<br.>
 +
PBS run to check for background fluorescence.  Sample was run three times, 200s each to eliminate noise.<br.>
 +
[[Image:Avg_125pM.png]]<br.>
 +
<br.>
 +
[[Image:Avg_100pM.png]]<br.>
 +
<br.>
 +
[[Image:Avg_75pM.png]]<br.>
 +
<br.>
 +
[[Image:Avg_50pM.png]]<br.>
 +
<br.>
 +
[[Image:Avg_25pM.png]]<br.>
 +
All DNA/MB samples run three times, for 500s.<br.>
 +
<br.>
 +
Calibration Graph:<br.>
 +
[[Image:2013_0705_calibration.PNG]]

Current revision

FCS Measurements Main project page
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July 5, 2013

FCS Sample Prep

  • Green fluorescent beads
    • 10000x diluted, run 3x for 90s
  • PBS
    • To check for background, run 3x for 200s
  • DNA/MB
    • 125/100pM, run 3x for 500s
    • 100/80pM, run 3x for 500s
    • 75/60pM, run 3x for 500s
    • 50/40pM, run 3x for 500s
    • 25/20pM, run 3x for 500s

FCS Data

Graphs generated using Igor Pro.

Image:Avg_Beads.png
10000x diluted green fluorescent beads were used to align the laser. Sample was run three times, for 90s each.

Image:Avg_PBS.png
PBS run to check for background fluorescence. Sample was run three times, 200s each to eliminate noise.
Image:Avg_125pM.png

Image:Avg_100pM.png

Image:Avg_75pM.png

Image:Avg_50pM.png

Image:Avg_25pM.png
All DNA/MB samples run three times, for 500s.

Calibration Graph:
Image:2013_0705_calibration.PNG



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