howto:lifetime-fitting_using_the_flim_script
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writingroom:lifetime-fitting_using_the_flim_script [2020/05/12 14:17] – ruckelshausen | howto:lifetime-fitting_using_the_flim_script [2020/05/13 07:37] – ↷ Links adapted because of a move operation ruckelshausen | ||
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- | ====== Lifetime-Fitting with the FLIM Analysis ====== | + | ====== Lifetime-Fitting with the FLIM Analysis |
===== Introduction ===== | ===== Introduction ===== | ||
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**Response: | **Response: | ||
The file '' | The file '' | ||
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**Note:**\\ | **Note:**\\ | ||
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* Open the " | * Open the " | ||
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A large window with three sections will appear, where you can set a threshold for an image analysis visually. | A large window with three sections will appear, where you can set a threshold for an image analysis visually. | ||
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**Note:**\\ | **Note:**\\ | ||
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* In the fourth box, "Decay Fitting", | * In the fourth box, "Decay Fitting", | ||
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* From " | * From " | ||
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* In this example it is obvious that the applied single exponential reconvolution fit is not suited to fit the decay curve. Increase the "Model Parameters" | * In this example it is obvious that the applied single exponential reconvolution fit is not suited to fit the decay curve. Increase the "Model Parameters" | ||
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* Three components exponential decay seems to be suitable for this ROI. However we can try four exponential decay and see the result:\\ | * Three components exponential decay seems to be suitable for this ROI. However we can try four exponential decay and see the result:\\ | ||
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**Response: | **Response: | ||
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* Set the same limit for A2 and A3.\\ | * Set the same limit for A2 and A3.\\ | ||
* Uncheck the boxes for the individual lifetime components, Bkgr(Decay), | * Uncheck the boxes for the individual lifetime components, Bkgr(Decay), | ||
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* Click "FLIM Fit" in the upper panel.\\ | * Click "FLIM Fit" in the upper panel.\\ | ||
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* An updated lifetime frequency histogram on the upper right. The individual lifetime components are displayed as vertical bars, as they are not varied in the pixel-by-pixel fit.\\ | * An updated lifetime frequency histogram on the upper right. The individual lifetime components are displayed as vertical bars, as they are not varied in the pixel-by-pixel fit.\\ | ||
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* The three lifetime components can be displayed in a false color scale. Therefore switch the display to " | * The three lifetime components can be displayed in a false color scale. Therefore switch the display to " | ||
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**Note:**\\ | **Note:**\\ | ||
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* In RGB mode you can select suited color to your favorite parameter. This helps to distinguish the lifetime of each specious or compartment of the cell. For instance lifetime in nuclei is shown here:\\ | * In RGB mode you can select suited color to your favorite parameter. This helps to distinguish the lifetime of each specious or compartment of the cell. For instance lifetime in nuclei is shown here:\\ | ||
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* When the desired display is set, click "Save Result" | * When the desired display is set, click "Save Result" |
howto/lifetime-fitting_using_the_flim_script.txt · Last modified: 2023/08/03 14:16 by lan