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Hi Hanna,

Since you use WLL on SP8, did you play around the AOBM (excitation selected
from WLL) and the AOBS (emission window) to see how the IRFs change at
different excitation/emission wavelengths?  This may tell you something for
what is going on.  Good luck with your measurements.

Sheng


On Fri, Nov 20, 2015 at 2:14 AM, Hanna Sas Nowosielska <
[log in to unmask]> wrote:

> *****
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> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Thank you for all suggestions, unfortunately we cannot use scatter light to
> get IRF. There is some problem either with our WLL laser or with our
> microscope system (Leica SP8) and when using scatter light we don’t obtain
> specific IRF. Instead we get multiple picks of different height and size.
> We talked to service but they’re still trying to find the source of the
> problem.
>
>
> Hanna
>
> 2015-11-20 3:25 GMT+01:00 yuansheng.sun <[log in to unmask]>:
>
> > you can try to use the cream coffee mate solution to record the laser
> > scatter directly.
> >
> > Sheng
> >
> >
> > Sent from my T-Mobile 4G LTE Device
> >
> >
> > -------- Original message --------
> > From: Hanna Sas Nowosielska <[log in to unmask]>
> > Date:11/19/2015  6:49 AM  (GMT-06:00)
> > To: [log in to unmask]
> > Cc:
> > Subject: Measuring IRF in red channel
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Dear Colleagues,
> >
> >
> >
> > Could someone suggest me a fluorophore with lifetime short enough to
> > measure IRF in red channel (excitation: 520nm/ Detection : 565 – 605)? I
> > would be very grateful for an advice.
> >
> >
> >
> > Best regards,
> >
> > Hanna
> >
> >
> >
> > --
> > Hanna Sas-Nowosielska
> >
> > Laboratory of Imaging Tissue Structure and Function
> > Nencki Institute of Experimental Biology
> > Warsaw, Poland
> >
>
>
>
> --
> Hanna Sas-Nowosielska
>
> Laboratory of Imaging Tissue Structure and Function
> Nencki Institute of Experimental Biology
> Warsaw, Poland
>