Biomod/2011/TUM/TNT/LabbookA/2011/09/30

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30th Sep 2011

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Single Molecule Fluorescence Microscopy Measurement of BM14_5/20 and BM21_172/173

  • As described on 29th Sept 2011, suitable origami concentrations for single molecule fluorescence microscopy needed to be found. Therefore the sample BM14_5/20 without any DNA binder diluted 1:1000, 1:5000 and 1:10000 were measured first.
  • It showed that a dilution of 1:5000 results in an acceptable peak density (see picture below).

Image:Capture.PNG
Fig.1:
Single molecule fluorescence microscopy picture of BM14_5/20 without DNA binder diluted 1:5000, excited with green laser at 532nm and detected in green camera


  • The framerate was set to 20 pictures per second and the number of frames was 500 per video.
  • The FRET efficiencies were calculated with Matlab (see explanation Evaluation of TIRF data).
  • A histogram of the calculated FRET efficiencies can be seen below at the example of BM14_5/20 with 4.8 μM of spermine.

Image:BM14_spermin_Histogramm.PNG
Fig.2:
Histogram of FRET efficiencies of the sample BM14_5/20 with 4.8 μM of spermine