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howto:calculate_and_fit_fcs_traces_with_the_fcs_script

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howto:calculate_and_fit_fcs_traces_with_the_fcs_script [2014/06/18 09:21] adminhowto:calculate_and_fit_fcs_traces_with_the_fcs_script [2014/06/18 09:22] admin
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 ===== Background Information ===== ===== Background Information =====
  
-ATTO488 (ATTO-Tec, Germany) is a suited calibration dye to calibrate the confocal microscope at ~470 – 490 nm excitation and detection ~500 – 540 nm.\\ +  * ATTO488 (ATTO-Tec, Germany) is a suited calibration dye to calibrate the confocal microscope at ~470 – 490 nm excitation and detection ~500 – 540 nm. 
-The diffusion coefficient of ATTO488 carboxilic acid has been measured to be 400 µm²/s at 25°C. Published diffusion coefficients of various fluorophores are summarized in the Technical Note "Absolute Diffusion Coefficients: Compilation of Reference Data for FCS Calibration" available from PicoQuant [see http://www.picoquant.com/images/uploads/page/files/7353/appnote_diffusioncoefficients.pdf ]. This list is extensive, but does not claim to be complete.\\ +  The diffusion coefficient of ATTO488 carboxilic acid has been measured to be 400 µm²/s at 25°C. Published diffusion coefficients of various fluorophores are summarized in the Technical Note "Absolute Diffusion Coefficients: Compilation of Reference Data for FCS Calibration" available from PicoQuant [see http://www.picoquant.com/images/uploads/page/files/7353/appnote_diffusioncoefficients.pdf ]. This list is extensive, but does not claim to be complete. 
-Note that the diffusion coefficients of all dyes are temperature dependent.\\ +  * **Note** that the diffusion coefficients of all dyes are temperature dependent. 
-FCS measurements are usually point measurements acquired with very sensitive detectors (typically SPAD or Hybrid PMT detectors).\\ +  FCS measurements are usually point measurements acquired with very sensitive detectors (typically SPAD or Hybrid PMT detectors). 
-FCS is a technique that can be used over a wide range of concentrations, but ideal FCS concentrations are usually between 1 nM and 20 nM.\\+  FCS is a technique that can be used over a wide range of concentrations, but ideal FCS concentrations are usually between 1 nM and 20 nM.
  
 ===== Step-by-Step Tutorial ===== ===== Step-by-Step Tutorial =====
howto/calculate_and_fit_fcs_traces_with_the_fcs_script.txt · Last modified: 2015/11/03 14:11 by admin