Search

You can find the results of your search below.

lifetime_fitting_using_the_flim_analysis
60 Hits, Last modified:
hows step-by-step, how the FLIM analysis of SymPhoTime 64 can be used to perform a lifetime fit and how to extract and read the results. In d... ile the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times of the photons after the laser pulse. When not d... which makes the color scale loaded by default sometimes unsuited for displaying the lifetime contrast in the actual sample. In this case, the ... s on the right of the plot.\\ **Lower left:** Lifetime fitting parameters. Here lifetime fitting models can be defined. Fitting is then ap
flim_fret_calculation_for_multi_exponential_donors
51 Hits, Last modified:
portant to use the amplitude weighted average lifetime. While the intensity weighted average lifetime corresponds to the real average arrival time of the photons and is therefore also close to the Fast lifetime in case of low background noise, the amplitude weighted average lifetime reflects the quenching of the donor due to the FR... ile the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times of the photons after the laser pulse. When not d... which makes the color scale loaded by default sometimes unsuited for displaying the lifetime contrast in the actual sample. In this case, the
lifetime-fitting_using_the_rapid_reconvolution_algorithm
44 Hits, Last modified:
tains now the fitted, intensity weighted mean lifetime, instead of the "fast Lifetime", based on the photon arrival time only. The Histogram on the right of the image con... d distribution of the intensity-weighted mean lifetime. The distributions for lifetime component 1 and lifetime component 2 appear as bars, as these parameters h... the result image after the fit with the fast lifetime image above, you will see that the lifetime distribution of the lifetime within a single bead is much more uniform, as eff... orrected for.\\ * If you want to export the lifetime distribution, open the context menu via right mou
intensity_time_trace_analysis
43 Hits, Last modified:
{{tag> tutorial howto time-trace analysis SymPhoTime}} ~~NOTOC~~ ======Intensity Time Trace Analysis====== ===== Summary =====... start the script ==== * Start [[products:SymPhoTime64|SymPhoTime 64]] software. * Open the "Samples" workspace v... ript_Image_4.png }} **Response:** The "Intensity Time Trace" script is applied to the file ''Cy5_immo_Lifetime_Trace.ptu''. Thereby, a new Window opens: {{ usin... Plotting options. - **Center/right:** Intensity time trace. As no time traces have been calculated yet, this area is sti
lifetime-fitting_using_the_flim_script
43 Hits, Last modified:
{{tag>howto tutorial FLIM analysis SymPhoTime MicroTime LSM_upgrade}} ====== Lifetime-Fitting Using the FLIM Analysis (updated for SymPhoTime V 2.5 and above) ====== ===== Introduction ​===== This tutorial shows step-by-step, how the FLIM script of SymPhoTime 64 (V2.5) can be used to perform a pixel-by-pixel lifetime fit of a FLIM image and how to extract and read t... ile the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times of the photons after the laser pulse. When not d
lifetime_fitting_using_the_tcpsc_fitting_script
41 Hits, Last modified:
{{tag>howto analysis lifetime tutorial SymPhoTime}} ~~NOTOC~~ ====== SymPhoTime Lifetime Fitting ====== ===== Summary ===== This tutorial shows step-by-ste... and start the script * Start [[products:SymPhoTime64|SymPhoTime 64]] software. * Open the "Samples" workspace v... SPC histogram it can be clearly seen that the lifetime of this dye is significantly shorter than the lifetime of ATTO655. {{ lifetime_fitting_using_the_tcpsc
flim-fret_calculation_for_single_exponential_donors
39 Hits, Last modified:
This tutorial shows step-by-step, how the "Lifetime FRET Image" analysis of [[products:SymPhoTime64|SymPhoTime 64]] can be used to calculate pixel-by-pixel the ... ce of FRET (τ<sub>D</sub>), while the shorter lifetime (τ<sub>DA</sub>) is varied. This lifetime is considered to be the donor lifetime with a FRET acceptor present. The FRET efficiency... ws a single peak, which is in contrast to the lifetime variations in the FLIM image. The explanation can... ram plots the amplitude fraction of the donor lifetime in presence of FRET. As the amplitude of a donor
visualizing_dynamics_using_the_multiframe-flim_script
36 Hits, Last modified:
ile the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times of the photons after the laser pulse. When not d... which makes the color scale loaded by default sometimes unsuited for displaying the lifetime contrast in the actual sample. In this case, the ... \\ **4: Decay Fitting Parameters**\\ Here the lifetime fitting model can be selected. Fitting is then applied to the lifetime graph(s) on the lower center ([[glossary:TCSPC]] ... cale by typing the limit values for the "Fast Lifetime [ns]" and "Events[cnts]". Additionally set the "Lifetime Histogram" range on the right hand.\\ {{ howto:f
roi_fitting_using_the_flim_script
33 Hits, Last modified:
[[glossary:flim|FLIM]] script of [[products:SymPhoTime64|SymPhoTime 64]] can be used to fit several regions of intere... start the script ==== * Start [[products:SymPhoTime64|SymPhoTime 64]] software. * Open the "Samples" workspace v... ile the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times of the photons after the laser pulse. When not d... which makes the color scale loaded by default sometimes unsuited for displaying the lifetime contrast in the actual sample. In this case, the
update
32 Hits, Last modified:
ile the color encodes the average "Fast FLIM" lifetime, i.e. the mean arrival times of the photons after the laser pulse. When not d... which makes the color scale loaded by default sometimes unsuited for displaying the lifetime contrast in the actual sample. In this case, the ... \\ **4: Decay Fitting Parameters**\\ Here the lifetime fitting model can be selected. Fitting is then applied to the lifetime graph(s) on the lower center ([[glossary:TCSPC]] ... cale by typing the limit values for the "Fast Lifetime [ns]" and "Events[cnts]". Additionally set the "Lifetime Histogram" range on the right hand. * Tick the
symphotime_tips_and_tricks
32 Hits, Last modified:
, which calculates the mean of the photon arrival times, you will notice that the lifetime values have partially negative values: {{fastfli... time) in the TCSPC fitting panel * the fast lifetime in an ideal system should equal the intensity weighted average lifetime [τ av. int.]. Usually, fitted lifetimes tend to be slightly shorter, as the background photons are excl... images obtained with PIE-excitation, the fast lifetimes are usually meaningless, as in this case the pho
visualizing_dynamics_with_the_multi_frame_flim_analysis
31 Hits, Last modified:
re (i.e. the piezo scanner in the [[products:MicroTime#MicroTime 200|MicroTime200]]), the image dimensions are automatically cor... ded, a procedure according to the tutorial: [[Lifetime fitting using the FLIM Analysis]] must be applied... 17 images have to be analyzed, which can be quite time consuming. Therefore, it can be helpful to limit
burst_duration_and_fcs_diffusion_time @howto:burst_duration_and_fcs_diffusion_time
29 Hits, Last modified:
= 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
separation_of_2_species_with
26 Hits, Last modified:
655 and ~1 ns for Cy5). In FLCS (Fluorescence Lifetime Correlation Spectroscopy), the lifetimes are used to create specific photon filters allow... start the script ==== * Start [[products:SymPhoTime64|SymPhoTime 64]] software. * Open the "Samples" workspace v... dye is analyzed that changes its fluorescence lifetime under certain conditions, a lifetime fit can be performed of the TCPSC histogram of th... LCS filters can be calculated from the fitted lifetime components. This approach called "LT (lifetime) fitting" can only be employed, if the two compon
calculate_ratiometric_single_pair_fret_distributions_using_the_pie-fret_script
21 Hits, Last modified:
{{tag> tutorial howto FRET PIE analysis time-trace SymPhoTime}} ~~NOTOC~~ ====== Calculate Ratiometric Single Pair FRET ... tart the script ==== * Start [[products:SymPhoTime64|SymPhoTime 64]] software. * Open the "Samples" workspace v... ** 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
calculate_ratiometric_single_pair_fret_distributions
19 Hits, Last modified:
how_to_measure_the_instrument_response_function_irf
19 Hits, Last modified:
avoid_pile_up_effect_in_flim_measurements
16 Hits, Last modified:
using_the_antibunching_script
15 Hits, Last modified:
diamond_nv_centers
14 Hits, Last modified:
calculate_and_fit_fcs_traces_with_the_fcs_script
14 Hits, Last modified:
antibunching_measurements
11 Hits, Last modified:
using_the_anisotropy_image_script
11 Hits, Last modified:
pattern_matching
10 Hits, Last modified:
determination_of_the_focal_width_with_the_focal
10 Hits, Last modified:
calculate_ratiometric_fret-images
10 Hits, Last modified:
calculate_fccs_trace_with_the_grouped_fcs_script
9 Hits, Last modified:
registering_new_scripts
8 Hits, Last modified:
howto
6 Hits, Last modified:
phasor_analysis
6 Hits, Last modified:
t3r_antibunching_-_slow_decay
5 Hits, Last modified:
check_overlap_of_different_color_confocal_volumes
5 Hits, Last modified:
align_beam_backreflection
5 Hits, Last modified:
measuring_quantum_yield
4 Hits, Last modified:
exchange_dichroic_mt200
4 Hits, Last modified:
data_file_import
3 Hits, Last modified:
reconvolution_fit
2 Hits, Last modified:
2ffcs
2 Hits, Last modified:
mt200everyday_alignment
1 Hits, Last modified:
mt200fcs
1 Hits, Last modified:
mt200fundamental_alignement
1 Hits, Last modified: