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- how_to_measure_the_instrument_response_function_irf
- t rate is much lower than the count rate during a fluorescence decay measurement. Diluting the scattering soluti... es. In order to avoid signal contamination by any fluorescence, the water must be very pure. ===== Using samp... lution. The solution is to acquire the IRF at the fluorescence wavelength, or at least spectrally closer to the fluorescence emission wavelength. ==== General recipe ====
- using_the_flcs_script_for_spectral_crosstalk_removal_via_flccs
- l shows step-by-step, how to calculate a spectral fluorescence lifetime cross-correlation (FLCCS) curve from a m... }} ===== Background Information ===== **Note:** Fluorescence Cross-Correlation Spectroscopy (FCCS) is a method... ission properties but they can also vary in their fluorescence lifetime.\\ To quantify the cross-correlation sig... ited with a pulsed and the other with a cw laser, fluorescence lifetime cross-correlation (FLCCS) is an ideal to
- diamond_nv_centers
- Emission filters therefor should be 650LP. ==== Fluorescence Lifetime ==== *Fluorescence Lifetime is approx. 12ns. ===== Measuring NV centers ===== [{{ howto:jablon... he nitrogene-vacancy in a diamond lattice. Bright fluorescence corresponds to the ^3E→^3A (m_s=0) transition}}] ... 0MHz - for finding the centers, later 10MHz (12ns fluorescence lifetime)) *excitation power: >7000 a.u. @ 4
- calculate_ratiometric_single_pair_fret_distributions
- itions. The script requires a spectrally resolved fluorescence time trace. As a result, the distribution of FRET... * The script requires a time trace containing the fluorescence of two spectrally separated channels, one channel mainly detecting the fluorescence of the donor dye and the other channel detecting mainly the fluorescence of the acceptor dye.\\ The time traces can be acq
- calculate_ratiometric_single_pair_fret_distributions_using_the_pie-fret_script
- (PIE). The script requires a spectrally resolved fluorescence time trace, in which donor and acceptor have been... * The script requires a time trace containing the fluorescence of two spectrally separated channels, one channel mainly detecting the fluorescence of the donor dye and the other channel detecting mainly the fluorescence of the acceptor dye.\\ The time traces can be acq
- using_the_anisotropy_image_script
- early polarized excitation and two detectors. The fluorescence should have been splitted with a polarizing beams... etectors. Consequently, one detector receives the fluorescence with a polarization parallel to the excitation li... sis and processing options. - **Upper center:** Fluorescence intensity image, displayed in false color scale. The perpendicular and parallel fluorescence intensities measured with the two detectors are d
- antibunching_measurements
- set laser repetition rate in order to allow full fluorescence to fully decay or highest possible rate or cw-exc... er resolvability of the dip, disadvantage is that fluorescence does not decay completely during the pulses. (Com... ching trace you will need at least a trace of 30s fluorescence emission with a minimum mean countrate of 10 kcps... //Counting constituents in molecular complexes by fluorescence photon antibunching IEEE Journal of Selected Topi
- calculate_fccs_trace_with_the_grouped_fcs_script
- 3E0?large }} ===== Background Information ===== Fluorescence Cross-Correlation Spectroscopy (FCCS) is a method... emission properties. They can also vary in their fluorescence lifetime, for more information please check the tutorials for Fluorescence Lifetime Correlation Spectroscopy (FLCS).\\ To qu... select the complete TCSPC window to avoid loosing fluorescence photons. * For channel A (= detector 1), mov
- flim_fret_calculation_for_multi_exponential_donors
- e of the background photons in the areas where no fluorescence is detected, is randomly spread over the TCSPC wi... , the scale has to be adapted.\\ **Upper right:** Fluorescence Lifetime Histogram. Here the frequency of photon ... e of the background photons in the areas where no fluorescence is detected, is randomly spread over the TCSPC wi... , the scale has to be adapted.\\ **Upper right:** Fluorescence FRET Efficiency Histogram. As the FRET efficiency
- separation_of_2_species_with
- al processes, i.e. cis/trans isomerization.\\ The fluorescence lifetime of both dyes differ significantly (~1.8 ns for ATTO655 and ~1 ns for Cy5). In FLCS (Fluorescence Lifetime Correlation Spectroscopy), the lifetimes... omponents.\\ Since the dye separation is based on fluorescence lifetime measurements via [[glossary:tcspc|TCSPC]... available, or a dye is analyzed that changes its fluorescence lifetime under certain conditions, a lifetime fit
- using_the_antibunching_script
- ep, how to calculate an antibunching curve from a fluorescence time trace of a single emitter, in this example a... *Note:** For calculating an antibunching curve, a fluorescence time trace has to be recorded.\\ To separate the fluorescence, the emission light has to be split onto two dete... culated. **Note:** The different peaks show the fluorescence of the second detector after each laser pulse, th
- visualizing_dynamics_with_the_multi_frame_flim_analysis
- e of the background photons in the areas where no fluorescence is detected, is randomly spread over the [[glossa... e scale has to be adapted.\\ - **Upper right:** Fluorescence Lifetime Histogram. Here the frequency of photon ... e selected for analysis. In this measurement, the fluorescence was separated by polarization, therefore, photons... ibed there in an analogous fashion:\\ * Fit the fluorescence decay and determine the fitting model and paramet
- flim-fret_calculation_for_single_exponential_donors
- uploads/page/files/7350/appnote_flim_overview.pdf|Fluorescence Lifetime Imaging (FLIM) in Confocal Microscopy Ap... e of the background photons in the areas where no fluorescence is detected, is randomly spread over the TCSPC wi... , the scale has to be adapted. - **Upper right: Fluorescence FRET Efficiency Histogram.** As the FRET efficien
- lifetime-fitting_using_the_flim_script
- e of the background photons in the areas where no fluorescence is detected, is randomly spread over the TCSPC wi... . In this case, the scale can be adapted.\\ **3: Fluorescence Lifetime Histogram**\\ Here the frequency of phot... eeded, if the pulse frequency is too high for the fluorescence decay to decrease to the background level before
- visualizing_dynamics_using_the_multiframe-flim_script
- h can report membrane tension changes through its fluorescence lifetime changes. This has been published 2018 in... e of the background photons in the areas where no fluorescence is detected, is randomly spread over the TCSPC wi... . In this case, the scale can be adapted.\\ **3: Fluorescence Lifetime Histogram**\\ Here the frequency of phot