User Tools

Site Tools


howto:calculate_and_fit_fcs_traces_with_the_fcs_script

Differences

This shows you the differences between two versions of the page.

Link to this comparison view

Both sides previous revision Previous revision
howto:calculate_and_fit_fcs_traces_with_the_fcs_script [2014/06/18 11:22]
admin
howto:calculate_and_fit_fcs_traces_with_the_fcs_script [2015/11/03 15:11] (current)
admin
Line 17: Line 17:
   * 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 ​[[glossary:SPAD]] or [[glossary: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.
  
Line 65: Line 65:
 {{ calculate_and_fit_fcs_traces_with_the_fcs_script_Image_7.png?​600 }} {{ calculate_and_fit_fcs_traces_with_the_fcs_script_Image_7.png?​600 }}
  
-**Note:** A possibility to remove the afterpulsing when working with cw excitation is splitting the light onto two detectors and calculate a cross correlation. Some detectors (mainly Hybrid ​PMTs) also have so low afterpulsing that the problem does not occur.+**Note:** A possibility to remove the afterpulsing when working with cw excitation is splitting the light onto two detectors and calculate a cross correlation. Some detectors (mainly ​[[glossary:Hybrid ​PMT]]s) also have so low afterpulsing that the problem does not occur.
  
   * If the sample has beenexcited with cw light, we would now save the data by clicking "Save Result"​ and press afterwards "​Transfer to Fit" (both in the "​File"​ dropdown panel). As our sample has been excited with a pulsed laser, we can apply one additional step to increase data quality.   * If the sample has beenexcited with cw light, we would now save the data by clicking "Save Result"​ and press afterwards "​Transfer to Fit" (both in the "​File"​ dropdown panel). As our sample has been excited with a pulsed laser, we can apply one additional step to increase data quality.
howto/calculate_and_fit_fcs_traces_with_the_fcs_script.txt · Last modified: 2015/11/03 15:11 by admin