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- calculate_and_fit_fcs_traces_with_the_fcs_script
- * The first decreasing part is due to detector afterpulsing and an unwanted artifact from the detector... png?600 }} **Note:** A possibility to remove the afterpulsing when working with cw excitation is splitti... mainly [[glossary:Hybrid PMT]]s) also have so low afterpulsing that the problem does not occur. * If t... save the data by clicking "Save Result" and press afterwards "Transfer to Fit" (both in the "File" dropdo
- antibunching_measurements
- are however artifacts caused by photons emitted (after the detection of a genuine photon) by one of the ... nd SPAD. The distance of the expected dip and the afterflash peak is characteristic for the setup. To get rid of the afterflash peak, it has been proven useful to introduce... ectly in front of each [[Glossary:SPAD]] E.g. the afterflash of MPD SPADs for Atto488 (485 nm excitation,
- calculate_ratiometric_single_pair_fret_distributions_using_the_pie-fret_script
- . The second area shows the fluorescence response after the second pulse (at 635 nm), which was used to e... ph is automatically assigned to the donor channel after donor excitation (detector 2). The second graph shows the acceptor trace after donor excitation – automatically assigned to channel 1. The third graph shows the acceptor trace after acceptor excitation, and the last graph shows the
- symphotime_tips_and_tricks
- e acquisition (you cannot change this information after the acquisition) More detailed information can b... y inside the ''.ptu'' file and cannot be modified after the measurement (see <imgref image1>). ==== Re... </WRAP>| * some parameters are only available after performing a FLIM-Fit (i.e. the different lifetim... "FastLT" along the line is displayed. \\ {{:howto:after_selection_line_profile.jpg?350|}} Note: The
- visualizing_dynamics_using_the_multiframe-flim_script
- etime, i.e. the mean arrival times of the photons after the laser pulse. When not defined otherwise, inte... the TCSPC decay window. {{ howto:rois_and_decays_afterbinning_2.png |}} ==== Perform an Initial Lifetim... ponse:**\\ The average lifetime, you can see that after adding the sugar, has been dropped gradually abou... more arrays depending on the fitting model used. After removal of the header and the unwanted array, ind
- lifetime-fitting_using_the_rapid_reconvolution_algorithm
- etime, i.e. the mean arrival times of the photons after the laser pulse. When not defined otherwise, inte... here is still a bit of deviation in the residuals after the IRF in the decay window. This however can be ... oi1.png |}} * If you compare the result image after the fit with the fast lifetime image above, you w
- roi_fitting_using_the_flim_script
- etime, i.e. the mean arrival times of the photons after the laser pulse. When not defined otherwise, inte... a second array encodes the Fast Lifetime values. After removal of the header and the unwanted array, ind... ROI 2\\ * Keep the cursor over the graph, and after a right mouse click, select from the context menu
- using_the_flcs_script_for_spectral_crosstalk_removal_via_flccs
- e two patterns: one for the fluorescence response after pulsed excitation (e.g. for the green fluorescent dye) and one for the fluorescence response after cw excitation.\\ After these TCSPC patterns have been generated, the green spectral channel using the pu
- calculate_fccs_trace_with_the_grouped_fcs_script
- g part of the autocorrelations is due to detector afterpulsing, an unwanted artifact from the detector. As the afterpulsing is not correlated between both detectors,
- calibrate_the_confocal_volume_for_fcs_using_the_fcs_calibration_script
- e background in the TCPSC window. As the detector afterpulsing is not correlated with the laser pulse on ... ime range of interest, photons caused by detector afterpulsing behave similar as background photons. *
- data_file_import
- ll end up in dedicated ''.etc'' data containers. After importing ''Decay_Coumarin_6.phu'', the workspace... ike this: {{ :howto:et2_workspace1.png |}} \\ \\ After expanding the data container: {{ :howto:et2_works
- flim_fret_calculation_for_multi_exponential_donors
- etime, i.e. the mean arrival times of the photons after the laser pulse. When not defined otherwise, inte... etime, i.e. the mean arrival times of the photons after the laser pulse. When not defined otherwise, inte
- how_to_measure_the_instrument_response_function_irf
- torage: <code> keep at ~ 4°C, renew the solution after one month </code> Spectra: <code> excitation fr... ctrometers, the IRF is usually measured before or after each measurement. In microscopy-applications, th
- lifetime_fitting_using_the_flim_analysis
- etime, i.e. the mean arrival times of the photons after the laser pulse. When not defined otherwise, inte... he pixel decay in grey, the background before and after the decay is practically 0.Therefore, we can also
- lifetime-fitting_using_the_flim_script
- etime, i.e. the mean arrival times of the photons after the laser pulse. When not defined otherwise, inte... he pixel decay in grey, the background before and after the decay is practically 0.Therefore, we can also