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- separation_of_2_species_with
- calculate separate autocorrelation curves for the two components of a mixture of ATTO655 and Cy5 based ... large }} ===== Background Information ===== The two fluorophores ATTO655 (ATTO-Tec, Germany) and Cy5 ... nt. The aim of this script is to demonstrate, how two FCS traces are calculated from the same dataset, ... In this file, the light was evenly splitted onto two detectors. Therefore, select both detectors in th
- calculate_ratiometric_single_pair_fret_distributions_using_the_pie-fret_script
- uires a time trace containing the fluorescence of two spectrally separated channels, one channel mainly... ) analysis of the script, pulsed excitation using two lasers is required, one laser exciting the donor,... aser driver PDL828 “Sepia II” in combination with two pulsed diode lasers. \\ ===== Step-by-Step Tut... ngs" unchanged and press "Calculate". **Note:** Two channels need to be active in order to calculate
- pattern_matching
- 600 }} **Response:** The image now clearly shows two populations of different lifetimes. This means, that probably two patterns are sufficient to fit the image. **Note... shows that the image can be readily described by two different decay patterns belonging to two different species, the FRET and non-FRET populati... terns for the different regions. * Extract the two patterns as described above and add them to the l
- flim_fret_calculation_for_multi_exponential_donors
- xample image used is an image showing four cells, two of them only transfected with the CENP-B-Cerulean complex, the other two transfected with both constructs. Therefore, the ... ted. **Note:**\\ Now four cells are visible, the two nuclei on the left have longer Fast lifetimes then the other two. These are the cells transfected only with the do... s, which is expected, as in the image one can see two cells with donor and acceptor construct and two cells without. For the cells with the FRET donor
- lifetime-fitting_using_the_rapid_reconvolution_algorithm
- each of these parameters individually. There are two ways to define the new threshold. You can use the... ount rates there is an increasing likelihood that two or more such detector pulses will overlap and mer... constant of the systematic errors. * Try last two steps with the ROI 1. {{ howto:monoexpo_roi1.pn... on is not sufficient. Therefore, we carry on with two components. {{ howto:biexpo_roi1.png |}} **
- calculate_ratiometric_single_pair_fret_distributions
- r where the acceptor was photobleached. The other two peaks are due to two different conformations of this RNA oligonucleoti
- using_the_flcs_script_for_spectral_crosstalk_removal_via_flccs
- ethod to determine molecular interactions between two molecules labeled with different dyes. Usually, the dyes attached to the two molecules differ in their absorption and emission... oomlight.\\ Therefore, it is possible to generate two patterns: one for the fluorescence response after... ins a FCS measurement of a FCCS negative control (two DNA oligonucleotides labeled with a green and a r... ting values are added to the Fitting table. * Two FLCS filters are generated: The pattern generated
- data_file_import
- native TimeHarp260 software]]. This file contains two histograms: an [[howto:how_to_measure_the_instrum... measurement and a decay measurement are stored in two separate ''.phu'' files. This is no problem, but ... data container: {{ :howto:et2_workspace2.png |}} Two histograms have been imported. They are shown as ... \ \\ Unlike ''Decay_Coumarin_6.phu'' containing two histograms, both ''Anthracene_EtOH.phu'' and ''IR
- intensity_time_trace_analysis
- nse:** As activated under "Select data channels", two intensity plots are calculated and plotted according to the two selected detector channels. {{ using_the_intensit... trace marked in green in the upper trace for the two channels.\\ **On the right**, photon counting his
- calculate_ratiometric_fret-images
- fret.pdf]].\\ The example image shows four cells, two of them were only transfected with CENP-B-Cerulean, the other two contain both constructs. The same image is also u... culate". **Response:**\\ * Automatically, the two active detector channels are assigned to the dono... sfected with the donor construct, while the other two cells are transfected with both constructs. The f
- calculate_fccs_trace_with_the_grouped_fcs_script
- ethod to determine molecular interactions between two molecules labeled with different dyes. Usually, the dyes attached to the two molecules of interest differ in their absorption ... , a negative calibration probe, i.e. a mixture of two single labeled compounds needs to be measured. To
- update
- tubul_magicwand20_3.png |}} * Repeat the last two steps in order to select the cell nuclei as the l... del should be used for ROI 0.\\ * Try the last two steps with "ROI 1".\\ {{ howto:tubul_3expo_3.png... {{ howto:showall.png |}} * Uncheck the first two datasets on top of the parameter table and press
- 2ffcs
- s-correlation between the occurrence of events in two different detection volumes is used to introduce ... somewhat sacrificing background rejection. === Two Foci === The distance between the foci in two-focus FCS is determined by the Nomarski prism. Usually
- burst_duration_and_fcs_diffusion_time @howto:burst_duration_and_fcs_diffusion_time
- detection volume is commonly characterized using two different observables: the diffusion time extract... full mathematical formalism. We then compare the two observables and establish their relationship unde... $t_D$. There is no universal ordering between the two quantities, whereas $t_D$ is largely insensitive
- using_the_antibunching_script
- rescence, the emission light has to be split onto two detectors with very high quantum yield (e.g. SPAD... center in the focus of the instrument. The other two files demonstrate changes of the antibunching cur... of the size of the other peaks, corresponding to two emitters in the confocal volume. * For the file