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lifetime_fitting_using_the_flim_analysis @howto
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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... s on the right of the plot.\\ **Lower left:** Lifetime fitting parameters. Here lifetime fitting models can be defined. Fitting is then ap... gging the limit bars in the color scale "Fast Lifetime" histogram or by considering the peaks of the "Lifetime Histogram". Entering fixed values has the advanta... now the calculated intensity weighted average lifetime "tau_Av_Int" is displayed, in contrast to the "Fast Lifetime", which is just the mean arrival time and contain
flim_fret_calculation_for_multi_exponential_donors @howto
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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... nential_donors_Image_3.png }} * Start the "Lifetime FRET Image" script by clicking on "Start". {{ fli... _donors_Image_25.png }} **Response:**\\ The “Lifetime FRET Image” script is applied to the file ''CENP-
software
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te software suite for acquisition and analysis of time resolved fluorescence data. SymPhoTime features a powerful scripting language that allow... EasyTau is the systems software of PicoQuants time resolved spectrometer [[products:FluoTime 300]]. EasyTau features a scripting language to a... ware designed for the phasor plot analysis of raw Time-Correlated Single Photon Counting (TCSPC) Fluorescence Lifetime Imaging Microscopy (FLIM) data. Developed in Pyth... nce-Tools/tttrlib|tttrlib]] to read and correlate time-tagged time resolved (TTTR) files. *[[https://github.com/y
lifetime-fitting_using_the_rapid_reconvolution_algorithm @howto
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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 @howto
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{{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 @howto
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{{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... \\ **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]]
lifetime_fitting_using_the_tcpsc_fitting_script @howto
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{{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 @howto
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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
fluorescence_correlation_spectroscopy-_a_short_introduction @general
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{{tag>MicroTime LSM Theory SymPhoTime FCS Correlation}} ====== Introduction to Fluorescence Correlation Spectroscopy ======... e mean product of the fluorescence intensity F at time t with the fluuorescence intensity after a lag time τ divided by the mean fluorescence intensity squa... interaction studies in cell membranes" * **Lifetime-filtered FCS** \\ Ishii et al., 2015, "Lifetime-Weighted FCS and 2D FLCS:Advanced Application of Time-Tagged TCSPC" \\ Gregor and Enderlein,2007 "Time Resolved methods in biophysics. 3. Fluorescence L
visualizing_dynamics_using_the_multiframe-flim_script @howto
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\\ **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... whether you have the scalebar active in the SymPhoTime. When measuring with a scanner controlled by SymPhoTime, e.g. a MicroTime system, or you transfer the image size informatio... . If your scanner is not controlled via the SymPhoTime, and no handshake transfers the correct image siz
roi_fitting_using_the_flim_script @howto
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[[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... the right of the plot.\\ - **Lower left:** Lifetime fitting parameters. Here lifetime fitting models can be defined. Fitting is then ap... gging the limit bars in the color scale "Fast Lifetime" histogram or by considering the peaks of the "Lifetime Histogram". Entering fixed values has the advanta
update @howto
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\\ **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 ... p, as an example, we mark an ROI with similar lifetime range using the "Magic Wand" tool. The sensitivity of the tool to lifetime differences can be adjusted in a box below the in... \\ The parts of the image with a similar fast lifetime range will be selected and the lifetime histogram will get updated accordingly. If you wa
visualizing_dynamics_with_the_multi_frame_flim_analysis @howto
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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
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= 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
some_origins_of_multiexponetial_decays_for_single_dyes
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s in fact the product of the rotation correlation time and the fluorescence lifetime, divided by their sum). And they also know that m... action relative the the original fluorescence lifetime, the observed decay time measured with a TCSPC spectrometer may be single-... will lead to a multiexponential decay. The ns lifetime corresponds to the intrinsic fluorescence lifetime of the fluorophore, whereas the ultrafast compone... tiexponential decays with positive (intrinsic lifetime, ns) and negative (rate of shift, ps) pre-exponen
symphotime_tips_and_tricks @howto
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t3-mode @glossary
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create_time_gated_image
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separation_of_2_species_with @howto
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microtime @products
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avoid_pile_up_effect_in_flim_measurements @howto
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t2-mode @glossary
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configuring_symphotime64_after_installation @support
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diamond_nv_centers @howto
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applications
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using_the_antibunching_script @howto
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measurement_hardware_instrumentation
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calculate_and_fit_fcs_traces_with_the_fcs_script @howto
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how_to_reinstall_symphotime
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pile-up_effect @glossary
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symphotime64 @software
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antibunching_measurements @howto
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pattern_matching @howto
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determination_of_the_focal_width_with_the_focal @howto
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start
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using_the_anisotropy_image_script @howto
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calculate_ratiometric_fret-images @howto
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tcspc_external_markers @support
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registering_new_scripts @howto
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tttr @glossary
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differential_count_rate @glossary
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bifl @glossary
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fast_lifetime @glossary
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video_tutorials
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dead_time @glossary
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mcs @glossary
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symphotime_32 @software
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lifetime_component_decomposition_phasor_plot_analysis
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symphotime @software
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measuring_quantum_yield @howto
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t3r_antibunching_-_slow_decay @howto
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flim @glossary
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phasor_analysis @howto
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faq @faq
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align_beam_backreflection @howto
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marquardt-levenberg @glossary
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hydraharp_400 @products
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howto @howto
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howto @tag
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deconvolution @glossary
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tcspc @glossary
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syntax @wiki
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fast_flim @glossary
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community_news @general
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laser_intensity_response @faq
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exchange_dichroic_mt200 @howto
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bootstrap @glossary
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picoharp_300 @products
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convolution @glossary
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reconvolution @glossary
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reconvolution_fit @howto
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synchrotron_applications
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pie @glossary
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novaflim_license @faq
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2ffcs @howto
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fluorescence_lifetime @glossary
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least_squares @glossary
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spad @glossary
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mcp @glossary
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pre-histogrammed_image @glossary
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imprint
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tcspc_electronics @products
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fcs @glossary
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novaflim_calculated_irf @faq
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irf @glossary
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flimfit @software
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lsm710
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fastlt @glossary
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faq
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bdf @glossary
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residuals @glossary
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asymptotic_standard_errors @glossary
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