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- calculate_ratiometric_single_pair_fret_distributions_using_the_pie-fret_script @howto
- PhoTime}} ~~NOTOC~~ ====== Calculate Ratiometric Single Pair FRET Distributions Using PIE-FRET ====== ... p-by-step, how to calculate a FRET histogram from single [[glossary:fret|FRET]]pairs detected under single molecule conditions using pulsed interleaved excitation... eptor dye.\\ The time traces can be acquired from single molecule events in a diluted solution, where the
- calculate_ratiometric_single_pair_fret_distributions @howto
- PhoTime}} ~~NOTOC~~ ====== Calculate Ratiometric Single Pair FRET Distributions ====== ===== Summary ===... calculate a [[glossary:fret|FRET]] histogram from single FRET pairs detected under single molecule conditions. The script requires a spectrally resolved fluoresce... eptor dye.\\ The time traces can be acquired from single molecule events in a diluted solution, where the
- software
- and configuration with PicoQuant Time Correlated Single Photon Counting Instruments (TCSPC devices). It h... github.com/OpenSMFS|Open Tools for Solution-Based Single-Molecule Fluorescence Spectroscopy]]** * **FR... : software for burst analysis of freely-diffusing single-molecule FRET (smFRET) measurements including μs-... https://github.com/tritemio/PyBroMo|simulator for single molecule FRET experiments of freely diffusing par
- fluorescence_correlation_spectroscopy-_a_short_introduction @general
- == In this short article only the case of point, single color FCS with a confocal set up will be examined... be monitored whereas in ~µs range processes like singlet-triplet kinetics, photo-isomerization and proton... anifested in the auto-correlation function is the singlet-triplet kinetics. Triplet states are "dark" in t... housands times, cycling between the first excited singlet state S1 and the ground state S0. With a certain
- applications
- sis software]] [[howto:flim-fret_calculation_for_single_exponential_donors|FLIM-FRET Calculation for Single Exponential Donors]] [[howto:flim_fret_calculation_f... tric FRET-Images]] [[howto:calculate_ratiometric_single_pair_fret_distributions|Calculate ratiometric single pair FRET distributions]] [[howto:calculate_ratiome
- flim-fret_calculation_for_single_exponential_donors @howto
- e}} ~~NOTOC~~ ====== FLIM-FRET Calculation for Single Exponential Donors ====== ===== Summary ===== ... acceptor pair. If the FRET donor molecule has a single exponential decay kinetic, not only the FRET effi... fret|FRET]]. Several fluorescent proteins with a single exponential decay behavior have been reported, e.... Highlight the file ''FRET_GFP and mRFP.ptu'' by a single mouse click. {{ :howto:flim-fret-1expd_gret_gfp_a
- using_the_antibunching_script @howto
- unching curve from a fluorescence time trace of a single emitter, in this example a nanodiamond NV center,... l are measurements of NV centers which are stable single photon sources excitable at 530 nm. The script is used to process one file with only a single NV center in the focus of the instrument. The oth... .\\ Antibunching measurements can be performed on single stationary molecules or at very low concentration
- flim_fret_calculation_for_multi_exponential_donors @howto
- e file ''CENP-labelled _cells_for_FRET.ptu'' by a single mouse click. {{ flim-fret-calculation_for_multi-e... green (= dataset 0) and the TCSPC histogram of a single pixel in grey (= dataset 1). In red an estimation... ''CENP-labelled _cells_for_FRET.ptu'' again by a single mouse click. {{ flim-fret-calculation_for_multi-e... can only be calculated if the FRET donor dye has single exponential decay kinetics, changing the fitting
- update @howto
- ghlight the file ''Kidney_Cells_FLIM.ptu'' by a single mouse click. * Select the **"Analysis"** tab ... aset 0) and the [[glossary:TCSPC]] histogram of a single pixel in grey (= dataset 1). In red an estimation... eparated regions with a similar lifetime into one single ROI, as it's done here, you have to press "Shift"... to:inner_monoexpo_3.png |}} ** Response:**\\ A single exponential reconvolution fit is performed. In th
- some_origins_of_multiexponetial_decays_for_single_dyes
- ay time measured with a TCSPC spectrometer may be single- or multiexponential. Let us consider the react... ion could take place. The decay measured would be single exponential and coincident with $\tau_A$. //Case... ime measured in the spectral region of A would be single exponential, with decay time $\tau_1$. However $\... and hence the whole system could be treated as a single dye. The decay would be single exponential, being
- diamond_nv_centers @howto
- s.png?250x180|Figure3: fluorescence spectrum of a single NV center in diamond excited with 532 nm}}] ====... oto-physical stability makes the nv-centers ideal single emitter samples compared to single molecules the laser power to excite them has to be considerably higher... *when measuring the fluorescence intensity of a single NV center, the count rate should stay stable. The
- lifetime-fitting_using_the_flim_script @howto
- ghlight the file ''Kidney_Cells_FLIM.ptu'' by a single mouse click. * Select the **"Analysis"** tab ... aset 0) and the [[glossary:TCSPC]] histogram of a single pixel in grey (= dataset 1). In red an estimation... oexpo_roi0_kidney_3.png |}} ** Response:**\\ A single exponential reconvolution fit is performed. In th... * In this example it is obvious that the applied single exponential reconvolution fit is not suited to fi
- lifetime-fitting_using_the_rapid_reconvolution_algorithm @howto
- * Highlight the file ''MicroBeads.ptu'' by a single mouse click. * Select the **"Analysis"** tab... aset 0) and the [[glossary:TCSPC]] histogram of a single pixel in grey (= dataset 1). In red an estimation... { howto:monoexpo_roi0.png |}} **Response:**\\ A single exponential fit is performed. In the TCSPC window... ll as the IRF background and IRF Fit. * One the single pixel level, the background can be neglected, and
- visualizing_dynamics_using_the_multiframe-flim_script @howto
- file ''Hyperosmotic_Shock_MDCK_Cells.ptu'' by a single mouse click. * Select the **"Analysis"** tab ... aset 0) and the [[glossary:TCSPC]] histogram of a single pixel in grey (= dataset 1). In red an estimation... it_multiframe_2.png?300 |}} ** Response:**\\ A single exponential reconvolution fit is performed. In th... * In this example it is obvious that the applied single exponential reconvolution fit is not suited to fi
- lifetime_fitting_using_the_flim_analysis @howto
- * Highlight the file DaisyPollen_cells_FLIM by a single mouse click. {{ lifetime-fitting_using_the_flim-s... green (= dataset 0) and the TCSPC histogram of a single pixel in grey (= dataset 1). In red an estimation... it" (marked in orange). **Response:**\\ * A single exponential reconvolution fit is performed. In th... \\ In this example it is obvious that the applied single exponential reconvolution fit is not suited to fi