cy using the "Lifetime FRET Image" analysis.
{{ youtube>zHfGae_Pd8A?large }}
===== Background informa... mage_9.png }}
**Note:**\\
For more information about the different ROI selection tools, check the tuto... of importance. Therefore exclude the bright dots outside the nucleus of the cell on the lower right by... ously "CRTL" and draw rectangles around the spots outside the nucleus.
* **Response:** The marked reg
ow to measure na IRF on confocal microscopes.
{{youtube>cjSgCBe3ReM?large}}
\\
==== Low count rate ... s, and it is very likely to produce artefacts without allowing to tell apart artefacts and effects.
==... lso a remarkably short fluorescence lifetime of about 10 ps. This short lifetime does not depend on the
ne the lifetime of ATTO655 diluted in water.
{{ youtube>WupJkKpUVV8?large }}
===== Step-by-Step Tutor... e is usually a bit arbitrary.\\
For explanation about the fitting model and the used equations, click o
Ch.0.
-Disconnect SPAD 1 Signal from PHR800 Router.
-Connect SPAD 1 Signal to [[Products:Pico... is NIM compatible. If the SPAD provides only TTL output pulses, you need to use the SIA400 adapter.
... ors directly to the [[Products:PicoHarp 300]] without passing SPAD 2 through the router. **In this case, however, the router cable (DVI-cable) needs to be d
h a TiSa-laser with 80MHz repetition rate). Check out the rapidReconvolution tutorial for this case (ht... d_reconvolution_algorithm).\\
For explanations about the fitting model and the used equations, click o... **\\
For pixel-by-pixel fits, we are interested about how the lifetime parameters are distributed in th
erest are defined which are finally fitted.
{{ youtube>g6uyvLQW4nY?large }}
===== Step-by-Step =====... ry selection, the corresponding red dot is greyed out.
{{ roi_fitting_using_the_flim_script_Image_14.pn... image are highlighted, everything else is greyed out. The decay corresponding to ROI 2 contains now th... e is usually a bit arbitrary.\\
For explanation about the fitting model and the used equations, click o
or after changing a key element
of the optical layout, for example the major dichroic, or changing the ... ra as well as to the basic confocal unit and finally to the detectors.
{{ youtube>EnOWrsxq_Q8?large }}
id for systems equipped with a 3mm dichroic.
{{ youtube>I54GjmS0iMQ?large }}
If you are unsure about any of the steps please contact** http://www.picoquan
e]]).
* It is important that the “Laser Power” output is not changed at the PDL or SEPIA laser drive... scribed here can also be used for IRF-fitting without time gating. However, as these decays contain the... ly. Please check the different FLIM-related tutorials for more information about the fitting.
is usually a bit arbitrary.\\
For explanation about the fitting model and the used equations, click o... er adding the sugar, has been dropped gradually about 500ps.\\
* If an IRF has been measured, the fi... and can be imported e.g. in Phasor Plot analysis routines.
\\
* A window opens asking for a place an... hown in the SymPhoTime 64 window, e.g with or without scale bar (depending on whether you have the scal
he best detector cannot deliver infinitely narrow output pulses. At increasing count rates there
is an ... ing curve, residuals and X².
For explanation about the fitting model and the used equations, click o