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- intensity_time_trace_analysis
- lifetime_fitting_using_the_flim_analysis
- lifetime_fitting_using_the_tcpsc_fitting_script
- lifetime-fitting_using_the_flim_script
- lifetime-fitting_using_the_rapid_reconvolution_algorithm
- recording_a_fluorescence_lifetime_image_flim_stack_with_a_lsm_upgrade_kit_on_a_nikon_a1
- symphotime_tips_and_tricks
- burst_duration_and_fcs_diffusion_time
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- intensity_time_trace_analysis
- Plotting options. - **Center/right:** Intensity time trace. As no time traces have been calculated yet, this area is sti... space, select the file ''GUVs.ptu'' and apply the time trace script to that file. When the intensity time trace window opens, click "Calculate" using a bin... of 1 ms as set by default. {{ using_the_intensity_time_trace_script_Image_12.png?600 }} **Note:** When the intensity time trace of an image is displayed, the scanning resu
- burst_duration_and_fcs_diffusion_time @howto:burst_duration_and_fcs_diffusion_time
- = 4.2 Physical interpretation ==== The diffusion time $t_D$ is not the mean residence time of an individual molecule. Instead, it is a corre... entally different observables. The FCS diffusion time $t_D$ is a correlation-derived geometry time scale governed by quadratic spatial weighting of ... relation. Flow introduces an additional transport time scale that leaves the FCS diffusion time unchanged when modeled explicitly, while directly... ely confined to slow non-stationarity. Diffusion time and burst duration should therefore be regarded a
- calculate_ratiometric_single_pair_fret_distributions_using_the_pie-fret_script
- ** Fitting and analysis options\\ - **Center:** Time traces. As the time traces are not yet calculated, this graph is stil... , calculate the FCS curve and check the diffusion time to see the average residence time of the sample in the laser focus. {{ calculate_r... tion) conditions, which is visualized in the "Set Time Gate" area. In this example first a 530 nm pulse ... and acceptor channel were recorded with a slight time delay, therefore two peaks can be seen in this ar
- calculate_ratiometric_single_pair_fret_distributions
- d analysis options.\\ - **Upper center/right:** Time traces. As the time traces are not calculated yet, this graph is stil... , calculate the FCS curve and check the diffusion time to see the average residence time of the sample in the laser focus. {{ calculate_r... and acceptor channel were recorded with a slight time delay, therefore two peaks can be seen in the gre... IE-FRET Script]]). **Response: **\\ * Three time traces are calculated and displayed in the window
- avoid_pile_up_effect_in_flim_measurements
- orrect average count rate.)) * At a pixel dwell time of 10µs each pixel is displayed as one point in the time trace. At a pixel dwell time of 1 µs the average of 10 pixels are displayed as one point in the time trace. * During the Test FLIM measurement the laser intensity is set th... s present. Procedure: * Calculate the exposure time per pixel: Multiply the number of frames with the pixel dwell time * Determine the number of photons via the data ... e the number of photons per pixel by the exposure time per pixel ===== Differential Count Rate ===== se
- how_to_measure_the_instrument_response_function_irf
- ved. Such a cocktail cannot be stored for a long time. Latest the KI will photochemically decompose. =... of course) any of the above mentioned fast decay time fluorophores and record their response. The signa... surement series, provided that the system has had time to warm up (~15min), and neither the repetition r... used as for the sample), and re-used later. Over time or upon changes of the excitation wavelength, the
- calculate_and_fit_fcs_traces_with_the_fcs_script
- page.\\ - **Upper center/right:** the intensity time trace. The display option can be changed using th... ing is not correlated with the laser pulse on the time - range of interest, photons caused by detector a... cs at a timescale short or equal to the diffusion time through the confocal volume. Examples for these d... It can be clearly seen that at short correlation time ranges, the curve does not fit the data well. Thi
- using_the_antibunching_script
- mber of simultaneously emitting molecules, a dead time free setup has to be employed for that time range. The SymPhoTime 64 script analyses data rec... citation may be used, but usually the measurement time with cw light is significantly larger. Therefore,... elp page.\\ - **Upper center/right:** Intensity time trace. The display can be changed using the "Trac... .\\ As expected for a single emitter, the peak at time 0 is practically missing. This corresponds to the
- how_to_work_with_the_instrument_response_function_irf
- ths of all contributions. The observed decay in a time domain measurement is a convolution of the 'real'... pdated.\\ 4. Select the decay range by Setting a `Time Gate` (move the red bars) to limit the decay the ... sed for the timegate. * In order to define the time range better, double click on the graph with the ... indow with you can enlarge in order to define the time gates better. When content, click o.k. * Deca
- lifetime_fitting_using_the_flim_analysis
- etch from minimum to maximum. As the mean arrival time of the background photons in the areas where no f... ad over the TCSPC window, the mean photon arrival time of the dark background is usually very long (up t... the decay in the TCSPC window. As a mean arrival time is calculated, it is possible that in dark region... the analysis. This also decreases the processing time. {{ lifetime-fitting_using_the_flim-script_Image
- flim_fret_calculation_for_multi_exponential_donors
- etch from minimum to maximum. As the mean arrival time of the background photons in the areas where no f... ad over the TCSPC window, the mean photon arrival time of the dark background is usually very long (up t... lifetime corresponds to the real average arrival time of the photons and is therefore also close to the... etch from minimum to maximum. As the mean arrival time of the background photons in the areas where no f
- lifetime-fitting_using_the_rapid_reconvolution_algorithm
- etch from minimum to maximum. As the mean arrival time of the background photons in the areas where no f... ad over the TCSPC window, the mean photon arrival time of the dark background is usually very long (with... exponentials that have a lifetime longer than the time constant of the systematic errors. * Try last ... the "fast Lifetime", based on the photon arrival time only. The Histogram on the right of the image con
- visualizing_dynamics_with_the_multi_frame_flim_analysis
- etch from minimum to maximum. As the mean arrival time of the background photons in the areas where no f... [glossary:TCSPC]] window, the mean photon arrival time of the dark background is usually very long (up t... and a (pixel) binning of 3 to decrease processing time and increase photon numbers per pixel. {{ visuali... 17 images have to be analyzed, which can be quite time consuming. Therefore, it can be helpful to limit
- diamond_nv_centers
- NV center, the count rate should stay stable. The time trace in figure 8 shows a reduction of the countr... er choose a NV-center that is isolated, measuring time of 120 s should be enough to yield an antibunchin... Hz rep rate., 2304.94 a.u.;4 ms/pixel 45 min rec. time!}}] [{{ howto:diamond_roi_lifetime.png?270x300|... : Antibunching measured on NV-center. Acquisition time 120 s, 1.3 N.A. oil immersion, 3534.98 a.u. 532 n
- lifetime-fitting_using_the_flim_script
- etch from minimum to maximum. As the mean arrival time of the background photons in the areas where no f... ad over the TCSPC window, the mean photon arrival time of the dark background is usually very long (with... pulse does not shift within the image acquisition time within the TCSPC window, therefore we fix the par... e "Fast Lifetime", which is just the mean arrival time and contains e.g. background photons.\\ * An up