cted with the donor construct by performing a ROI-fit with the FLIM-analysis. The amplitude weighted av... y lifetime and also to perform the pixel-by-pixel fit to calculate the FRET efficiency.
===== Step-by-... itting model.
{{ :howto:flim-fret-multiexpd_reconvfit.png?400 |}}
**Response:**\\
* In the TCSPC w... ameter table.
{{ :howto:flim-fret-multiexpd_reconvfit_response.png |}}
**Note:**\\
The software offers
hub.com/PicoQuant/EasyTau-Scripts
[[software:FluoFit]] \\
Decay curve analysis software
===== Open Sou... e-Molecule Spectroscopy]]
* [[software:PyCorrFit]] is a general-purpose FCS evaluation software t... ablished Zeiss ConfoCor3 ~.fcs file format. PyCorrFit comes with several built-in model functions, cove... g (DLS).
* [[http://www.dkfz.de/Macromol/quickfit|QuickFit 3.0]]: QuickFit 3 is our open-source dat
=====
This tutorial shows step-by-step, how to fit a selected model to a decay curve in order to get... As an example, a single exponential reconvolution fit is used to determine the lifetime of ATTO655 dilu... offers the possibility to use a n-exponential tailfit or a n-exponential reconvolution model fit. A tailfit can be used when the expected lifetimes are sign
can be used to perform a pixel-by-pixel lifetime fit of a FLIM image and how to extract and read the r... ited fitting model is chosen and a pixel-by-pixel fit is performed.
===== Step-by-step Tutorial ====... lly want to have in a pixel in order to perform a fit. If there are too few photons in a pixel, a decent fit result cannot be expected. It is also possible to
ympus FV1200 upgrade kit]]
[[howto:calculate_and_fit_fcs_traces_with_the_fcs_script|Calculate and fit FCS curves]]
[[howto:calibrate_the_confocal_volume_f
ducts:SymPhoTime64|SymPhoTime 64]] can be used to fit several regions of interest (ROIs), and how to ex... ution of the raw image.
==== Perform a Lifetime Fit on the Selected Regions (ROIs) ====
* In the d...
**Note:** The software offers the possibility to fit the data using a n-exponential tailfit or a n-exponential reconvolution fit. A tailfit can be used whe
w the FLIM script of SymPhoTime 64 can be used to fit several regions of interest (ROIs), and how to ex... ited fitting model is chosen and a pixel-by-pixel fit is performed.\\
===== Step-by-step Tutorial ===... orresponding to ''ROI 0'' will appear under decay fit in "Decay Fitting" panel.\\
**Note:**\\
For fin... n the list.
==== Perform an Initial Lifetime Fit on the Selected Regions (ROIs) ====
* In the
nning_2.png |}}
==== Perform an Initial Lifetime Fit on the Selected Regions (ROIs) ====
* In the ... rop-down list select "n-Exponential Reconvolution Fit".\\
{{ howto:nexpon_fitmodel.png?300 |}}
** Res... *Note:**\\
The software offers the possibility to fit the data using a n-exponential tail fit, n-exponential reconvolution or rapid reconvolution fit. A tai
is of SymPhoTime 64 is used to perform a lifetime fit and how to extract and read the results. A mixtur... dow and an extra gray decay on bottom right under Fit. Lifetime histogram and the preview FLIM image re... th ROI 0 and only one component (monoexponential) fit. Press "Initial Fit" (marked in orange).
{{ howto:monoexpo_roi0.png |}}
**Response:**\\
A single ex
===== Reconvolution fit =====
Select an ''.etc'' container that contains at least one IRF histogram an... d. The default fitting option is "Exponential Tailfit". In order to perform a reconvolution fit, we need to select and then assign a curve as an IRF and the... th IRF and decay curve time ranges. Pressing the "Fit Single" button performs a fully automated fit of
e are many options in this panel, not all of them fit into the visible area of the screen. Thus, a scro... des the Fast Lifetime values. If a pixel-by-pixel fit has been performed, there are more arrays dependi... ing_dynamics_Image_19.png }}
==== Perform a FLIM Fit ====
**Note:** If not only the Fast FLIM image should be exported, but also a pixel-by-pixel fit for the whole stack is needed, a procedure accord
ertain regions of the cytoplasm. A bi-exponential fit is performed in a larger region of interest to de... sence of [[glossary:fret|FRET]]. A pixel-by-pixel fit is performed in the image and in every pixel the ... 1expd_fittingmodel.png?400 |}}
* Click "Initial Fit" below the fitting parameter.
**Response:** The fit is performed to the ROI decay.
{{ :howto:flim-fre