[FLIMfit-users] FLIMfit and Time-Gated Data

[Munro, Ian]

Hi Romain 

I know that our group uses IRFs with 25ps resolution for gated data, as described at http://docs.flimfit.org/advanced.html#creating-a-library-irf
Can I ask how that relates to the process you describe?

Thanks and best wishes

Ian



> On 24 Jan 2017, at 16:25, R.F. Laine <rfl30 at cam.ac.uk> wrote:
> 
> Hi Susana,
> I also use Erythrosin B in waterfor my IRF measurement.
> Typically, to estimate the reference lifetime of Erythrosin B, I also acquire a sample with Rhodamine 6G and perform a monoexponential fit adding the reference lifetime as a fitted parameter.
> I know that Rhodamine 6G lifetime is ~4ns and I can check whether the fit is performed correctly. You can do this on a central ROI of your image.
> 
> You'll need to estimate the IRF shift beforehand (also by fitting it), but beware that fitting both IRF shift and reference lifetime at the same time makes FLIMfit sometimes crash (fatal error) in my hands.
> 
> I hope this helps.
> 
> Cheers,
> 
> Romain
> 
> 
> 
> 
> On 2017-01-24 15:36, Susana Silva wrote:
>> Dear All
>> I am working with a Time-Gated FLIM system based on a LaVision
>> intensified CCD camera capable of producing gates with widths ranging
>> from 200ps to 1000ps. I am trying to used FLIMfit to fit the acquired
>> data. Right now, I am trying to perform a monoexponential fit on a
>> dataset acquired from a green chroma slide with an IRF acquired in
>> fluorescence mode from a solution of erythrosine B in water (MiLiQi),
>> but I have some issues regarding the quality of the fit and the
>> reference lifetime I have to use in the IRF tab before fitting the
>> data. I read somewhere in this mailing list that the reference
>> lifetime should be estimated by a monoexponential tail fitting of the
>> IRF curve. However, this advice was to TCSPC data and not Time-Gated.
>> With Time-Gated IRF does the same procedure apply? If so, how can
>> eliminate the gate effect when using large gate widths such as 1000ps?
>> Best Regards,
>> Susana Silva
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> 
> -- 
> Dr. Romain Laine, PhD in Biophotonics
> Laser Analytics Group
> Department of Chemical Engineering and Biotechnology
> University of Cambridge
> New Museums Site
> Pembroke Street, Cambridge, CB2 3RA, UK
> T: (+44)1223330133
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