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howto:visualizing_dynamics_with_the_multi_frame_flim_analysis

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howto:visualizing_dynamics_with_the_multi_frame_flim_analysis [2015/11/03 14:31] adminhowto:visualizing_dynamics_with_the_multi_frame_flim_analysis [2015/11/03 14:33] admin
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 **Note:** The window contains five different regions:\\ **Note:** The window contains five different regions:\\
   - **Upper left:** Imaging analysis options. As there are many options in this panel, not all of them fit into the visible area of the screen. Thus, a scroll bar is generated to access all controls.\\   - **Upper left:** Imaging analysis options. As there are many options in this panel, not all of them fit into the visible area of the screen. Thus, a scroll bar is generated to access all controls.\\
-  - **Upper center:** [[glossary:Fast FLIM]] image displayed in false color scale. The brightness encodes the intensity while the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times of the photons after the laser pulse. When not defined otherwise, intensity and color scale stretch from minimum to maximum. As the mean arrival time of the background photons in the areas where no fluorescence is detected, is randomly spread over the TCSPC window, the mean photon arrival time of the dark background is usually very long (up to 1/2 of the TCSPC window), which makes the color scale loaded by default sometimes unsuited for displaying the lifetime contrast in the actual sample. In this case, the scale has to be adapted.\\+  - **Upper center:** [[glossary:Fast FLIM]] image displayed in false color scale. The brightness encodes the intensity while the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times of the photons after the laser pulse. When not defined otherwise, intensity and color scale stretch from minimum to maximum. As the mean arrival time of the background photons in the areas where no fluorescence is detected, is randomly spread over the [[glossary:TCSPC]] window, the mean photon arrival time of the dark background is usually very long (up to 1/2 of the [[glossary:TCSPC]] window), which makes the color scale loaded by default sometimes unsuited for displaying the lifetime contrast in the actual sample. In this case, the scale has to be adapted.\\
   - **Upper right:** Fluorescence Lifetime Histogram. Here the frequency of photon counts corresponding to the individual mean lifetimes is plotted. The settings of the plot can be adapted using the controls on the right of the plot.\\   - **Upper right:** Fluorescence Lifetime Histogram. Here the frequency of photon counts corresponding to the individual mean lifetimes is plotted. The settings of the plot can be adapted using the controls on the right of the plot.\\
-  - **Lower left:** Lifetime fitting parameters allowing to define the lifetime fitting models. Fitting is then applied to the lifetime graph(s) in the TCSPC histogram.\\ +  - **Lower left:** Lifetime fitting parameters allowing to define the lifetime fitting models. Fitting is then applied to the lifetime graph(s) in the [[glossary:TCSPC]] histogram.\\ 
-  - **Lower center/left:** TCSPC histogram of all photons in the image in green (= dataset 0) and the TCSPC histogram of a single pixel in grey (= dataset 1). In red an estimation of the instrument response function (= IRF) is shown. The IRF reconstruction is deducted from the rising edge of the TCSPC histogram.+  - **Lower center/left:** [[glossary:TCSPC]] histogram of all photons in the image in green (= dataset 0) and the [[glossary:TCSPC]] histogram of a single pixel in grey (= dataset 1). In red an estimation of the instrument response function (= [[glossary:IRF]]) is shown. The [[glossary:IRF]] reconstruction is deducted from the rising edge of the [[glossary:TCSPC]] histogram.
  
   * To enhance the lifetime contrast within the image, adapt the color scale from 0 to 15 ns by typing these values as limits for the Fast Lifetime color scale. Also adjust the intensity scale to 1 – 200 counts.   * To enhance the lifetime contrast within the image, adapt the color scale from 0 to 15 ns by typing these values as limits for the Fast Lifetime color scale. Also adjust the intensity scale to 1 – 200 counts.
howto/visualizing_dynamics_with_the_multi_frame_flim_analysis.txt · Last modified: 2020/05/13 09:39 by ruckelshausen