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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... ing_Image_4.png?600 }} **Note:** In the example, two peaks in the average lifetime (x-axis) can alread... Zoom into the //Decay Diversity Map// to see the two different populations and their different lifetim
- 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
- calculate_ratiometric_single_pair_fret_distributions
- uires a time trace containing the fluorescence of two spectrally separated channels, one channel mainly... ings" unchanged and press "Calculate". **Note:** Two channels need to be active in order to calculate ... ter mainly excited via FRET - was detected in the two different channels. In this trace, donor and acce... were recorded with a slight time delay, therefore two peaks can be seen in the green marked area. This
- 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 |}} **
- using_the_flcs_script_for_spectral_crosstalk_removal_via_flccs
- (FLCCS) curve from a measurement of a mixture of two labeled oligunucleotides moving independently fro... 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
- 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
- , its fluorescence was split by polarization onto two detectors. {{ using_the_intensity_time_trace_scr... Image_5.png?600 }} **Note:** The window contains two different regions:\\ - **Left:** Plotting optio... nse:** As activated under "Select data channels", two intensity plots are calculated and plotted according to the two selected detector channels. {{ using_the_intensit
- 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
- 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
- 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
- lifetime_fitting_using_the_tcpsc_fitting_script
- Image_5.png?600 }} **Note:** The window contains two different regions:\\ - **Left:** Fitting and an... splayed in the script. For illustration, mark the two files ''Cy5_diff_IRF+FLCS-pattern.ptu'' and ''ATT... . In our example, it is of course meaningless, as two different dyes were fitted. * Place the mouse
- 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
- using_the_anisotropy_image_script
- s recorded with linearly polarized excitation and two detectors. The fluorescence should have been splitted with a polarizing beamsplitter onto the two detectors. Consequently, one detector receives th... allel fluorescence intensities measured with the two detectors are displayed in green (perpendicular)