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- symphotime_tips_and_tricks
- ype Overview ===== ^Workspace: may contain these Filetypes ^ Content of the file ^ How to get such a file in the workspace ^ How to open/process^ |''.ptu''... ==== * double click on any ''.ptu'' (raw data) file opens a file viewer, in which you find information about the file type and acquisition settings <imgcaption image1... e result of a particular analysis script. Result files also contain a comment which can be displayed via the file menu: **File → Show Comment**. {{show_comment.png|}} Typica... Images can be drawn in 3 ways (for this, open the file ''FLIM_3_expon.pqres'', associated to the raw data file ''DaisyPollen_cells_FLIM.ptu'') * toggling betw
- lifetime_fitting_using_the_tcpsc_fitting_script
- marked in green, indicating that it is the active file. Under decay data all files are listed, the active file is always highlighted in green. **Note:** The file ''Cy5_diff_IRF+FLCS-pattern.ptu'' contains a lifeti... 0 }} **Response:** A window opens and asks for a file name and a folder to store the data, e.g. as ''fi... psc_fitting_script_Image_22.png }} * Store the file. You can open the data e.g. with the notepad func
- data_file_import
- reateopen.png | File menu }} Then select Import Files. A new dialog opens, where the data file is selected: {{ :howto:et2_fileimport.png | Selects the file to be imported }} The file selected above is a sample data file, ''Decay_Coumarin_6.phu'' installed during the installation of the [[https:... y measurement are stored in two separate ''.phu'' files. This is no problem, but then the file import procedure has to be repeated for each file
- calculate_ratiometric_single_pair_fret_distributions_using_the_pie-fret_script
- et_script_Image_14.png }} **Response:** A result file (''PIE_FRET_TimeTrace_1.pqres'') is stored under the raw data file (''Cy3+Cy5_diff_PIE-FRET.ptu''). {{ calculate_rat... mage_15.png }} * Later, clicking on the result file reopens the file in the same way as it was stored. * Now the nec... e used automatically also for procession of other files, click "File" -> "Save Defaults". {{ calculate_ratiometric_sin... nse:** A window opens to select the user settings file. Select the recently stored file. {{ calculate_ratiometric_single_pair_fret_distri
- separation_of_2_species_with
- r its contents, we rename it: ''Cy5-FLCS.pqres''. Files can be renamed by clicking slowly twice onto the file name and typing in the new name. {{ separation_of... ts contents, rename it as ''ATTO655-FLCS.pqres''. Files can be renamed by clicking slowly twice onto the file name and typing in the new name. {{ separation_of... result" to save this curve. Rename the generated file as "Cy5-FLCS". * These curves can now be fitted
- calculate_ratiometric_single_pair_fret_distributions
- ributions_Image_13.png }} **Response:** A result file (''FRET_TimeTrace_1.pqres'') is stored under the raw data file (''Cy3+Cy5_diff_PIE-FRET.ptu''). {{ calculate_rat... mage_14.png }} * Later, clicking on the result file reopens the file in the same way as it was stored. * Now the nec... e used automatically also for procession of other files, click "File" -> "Save Defaults". {{ calculate_ratiometric_sin... nse:** A window opens to select the user settings file. Select the recently stored file. {{ calculate_ratiometric_single_pair_fret_distri
- visualizing_dynamics_using_the_multiframe-flim_script
- image. - 1x1 bitmap exports the image as a bmp file with one pixel of the image being exported as one pixel in the image file. In the other bitmap export options, the image is... IN exports the image series as a series of binary files. The data format of these files is described on the SymPhoTime Online help. The ... Save Results".\\ **Response:**\\ A result file is generated which is assigned to the raw data file. Double click on this ''.pqres''-file opens it in the same way as it was stored.
- roi_fitting_using_the_flim_script
- mage.\\ **1x1 bitmap** exports the image as a bmp file with one pixel of the image being exported as one pixel in the image file. In the other bitmap export options, the image is... 00 }} **Response:** A window opens, asking for a file name to store the fitting values. The data are stored as a ''.dat'' file. {{ roi_fitting_using_the_flim_script_Image_26.pn... Image_28.png?241 }} **Response:**\\ * A result file (''FLIM.pqres'') is stored under the raw data file (''DaisyPollen_FLIM.ptu''). {{ roi_fitting_using_... mage_29.png }} * Later, clicking on the result file reopens the file in the same way as it was stored. **Note:**\\ Th
- flim_fret_calculation_for_multi_exponential_donors
- esult_bearb.png?400 |}} **Response:**\\ A result file (''FLIM.pqres'') is stored under the raw data file (''CENP-labelled_cells_for_FRET.ptu''). {{ :howto... _donors_Image_44.png }} **Response:**\\ A result file is saved and linked to the raw data file. {{ :howto:flim-fret-multiexpd_pqres_2.png?400 |}
- using_the_flcs_script_for_spectral_crosstalk_removal_via_flccs
- oval_via_flccs_Image_9.png?336 }} * Select the file ''IBA488_IBA547_unlinked_mix.ptu'' (the same file again) and click "OK". {{ using_the_flcs_script_f... **Response:** * The TCSPC histogram for this file appears in the "Pattern" graph in the upper cente... visible, as the pattern was created from the same file, which makes the fitting perfect. * Fitting v... the fluorescence decay is named according to the file (it corresponds to the filter for the pulsed exci
- calculate_fccs_trace_with_the_grouped_fcs_script
- open the corresponding help page. The grouped FCS file can calculate and handle multiple FCS curves, either from different files or several correlations from one file, as used h... '. **Response:**\\ The correlation curves of this file are calculated. The cross-correlation amplitude of this file is ~20% of the autocorrelations. {{ calculate_fc
- using_the_anisotropy_image_script
- ge_script_Image_16.png }} **Response:** A result file (''AnisotropyImage_1.pqres'') is stored under the raw data file (''Cy5_immo_FLIM_Po-Imaging.ptu''). {{ using_the_... mage_17.png }} * Later, clicking on the result file reopens the file in the same way as it was stored. \\ ==== Stor... nse:** A window opens to select the user settings file. Select the recently generated file. {{ using_the_anisotropy_image_script_Image_23.pn... now the "Anisotropy Imaging" script again on the file, the user defined G-factor and detector assignmen
- visualizing_dynamics_with_the_multi_frame_flim_analysis
- mage.\\ **1x1 bitmap** exports the image as a bmp file with one pixel of the image being exported as one pixel in the image file. In the other bitmap export options, the image is... _dynamics_Image_18.png }} **Response:** A result file is generated which is assigned to the raw data file. Double click on this ''.pqres'' file opens it in the same way as it was stored. {{ visualizing_dynamic
- intensity_time_trace_analysis
- setups. * In the sample workspace, select the file ''GUVs.ptu'' and apply the time trace script to that file. When the intensity time trace window opens, clic... . * In this way, the marker assignment for this file has been checked and the window can be closed. As we are not interested in the data itself, there is no need to store this file.
- calibrate_the_confocal_volume_for_fcs_using_the_fcs_calibration_script
- on_script_Image_11.png }} **Response:** A result file is generated and linked to the raw data file. {{ calibrate_the_confocal_volume_for_fcs_using_t... cript_Image_12.png }} ==== Use the generated FCS file for calibration of the confocal volume ==== * Highlight the newly generated FCS result file. {{ calibrate_the_confocal_volume_for_fcs_using_t... on_script_Image_23.png }} **Response:** A result file is generated and linked to the raw data. The fitt