g>tutorial howto analysis FLIM imaging FRET SymPhoTime}}
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====== FLIM-FRET Calculation for M...
This tutorial shows step-by-step, how the "Lifetime FRET Image" analysis of SymPhoTime 64 can be used to calculate pixel-by-pixel the average [[glossary:fr... average FRET-efficiency, first the donor only lifetime is determined using cells that have only been tra
ved.
Such a cocktail cannot be stored for a long time. Latest the KI will photochemically decompose.
=... of course) any of the above mentioned fast decay time fluorophores and record their response. The signa... surement series, provided that the system has had time to warm up (~15min), and neither the repetition r... used as for the sample), and re-used later. Over time or upon changes of the excitation wavelength, the
{{tag>howto analysis lifetime tutorial SymPhoTime}}
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====== SymPhoTime Lifetime Fitting ======
===== Summary =====
This tutorial shows step-by-step, how to fit a selecte
etectors slightly.
===== Antibunching using SymPhoTime =====
*with [[Products:PicoHarp 300]] in [[g... ossary:T3-mode]]
*not possible with SymPhoTime (32-bit version), because in SPT32 the Sync Divid... 400]] softwares as well as with [[software:SymPhoTime64]], where the sync divider can be disabled.
Usi... *measurements are possible with [[software:SymPhoTime64]]
==== Changing from Normal Acquisition (T3-mo
{{tag>howto tutorial FLIM analysis SymPhoTime MicroTime LSM_upgrade}}
====== Lifetime-Fitting Using the FLIM Analysis (updated for SymPhoTime V 2.5 and above) ======
===== Introduction ====
g> howto FLIM imaging ROI analysis tutorial SymPhoTime}}
~~NOTOC~~
====== ROI Fitting Using the FLIM A... [[glossary:flim|FLIM]] script of [[products:SymPhoTime64|SymPhoTime 64]] can be used to fit several regions of interest (ROIs), and how to extract and interpr... start the script ====
* Start [[products:SymPhoTime64|SymPhoTime 64]] software.
* Open the "Samples
{{tag>howto tutorial FLIM analysis SymPhoTime MT LSM_upgrade}}
====== ROI Fitting Using the FLIM Analysis (updated for SymPhoTime v2.5)======
===== Introduction =====
This tutoria... shows step-by-step, how the FLIM script of SymPhoTime 64 can be used to fit several regions of interest... file and start the analysis ====
* Start SymPhoTime 64 software (this tutorial uses version 2.5, ther
{{tag>howto MicroTime200 dichroic microscopy}}
====== How to exchange the main dichroic of the MicroTime 200 ======
This video serves as additional supp... or the "Exchange of the main dichroic of the MicroTime 200." section of the MicroTime 200 manual. It is only valid for systems equipped with a 3mm dichroic.
{{tag>howto IRF tutorial analysis MicroTime LSM_upgrade}}
====== How to Work with Instrument Response ... cts how a component with an infinitive short
lifetime is seen by the instrument. Of course, all compone... ths of all contributions.
The observed decay in a time domain measurement is a convolution of the 'real'... by-step analysis incl. A measured IRF using SymphoTime software: =====
1. First the measured IRF need
ging analysis dynamics tutorial LSM_upgrade SymPhoTime}}
====== Visualizing Dynamics Using the Multi Frame FLIM Analysis (updated for SymPhoTime v2.5 and above) ======
===== Introduction =====
... age containing many frames is treated here like a time-series. The example image shows the plasma membra... rane tension changes through its fluorescence lifetime changes. This has been published 2018 in nature:
{{tag>howto tutorial FLIM analysis SymPhoTime MT LSM_upgrade reconvolution fitting}}
====== Lifetime-Fitting Using the Rapid Reconvolution Model =====... IM]].
Here in detail the FLIM analysis of SymPhoTime 64 is used to perform a lifetime fit and how to extract and read the results. A mixture of Dragon Green
{{tag>howto MicroTime200 microscopy backreflection alignment }}
====== Image on the beam diagnostics cam... d for daily and fundamental alignment of the MicroTime 200. A detailed procedure is described in the user manual of MicroTime 200. Although MicroTime is extremely robust, we suggest to spend 5-10 minutes every day for verifying t
indow3.png |}}
\\
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If necessary, change the time axis limits by dragging the appropriate vertical ... re dedicated sliders for both IRF and decay curve time ranges. Pressing the "Fit Single" button performs
e -analysis -draft &nodate&nouser}}
===== SymPhoTime64 / Analysis =====
The SymPhoTime tutorials rely on the sample workspace downloadable [[http://www.pic... /products/category/fluorescence-spectrometers|FluoTime series spectrometers]] controlled with [[https://... s://www.picoquant.com/products/category/tcspc-and-time-tagging-modules|TimeHarp260, PicoHarp300, HydraHa
st arriving photon against a photon at the longer time, which will be an increasingly unlikely event bec... experiment restarts.
A simulation with 90ns dead-time period:
[{{howto:logf.png?320x240|Fig. 2: Simul... cale is in ns. The plateau is the [[glossary:dead time]]. The decay slope depends on the count rate.
Th... nst every photon.
It can be done using the SymPhoTime Software (http://www.picoquant.com/products/sw_mt