Measuring IRF in red channel

[Hanna_SN]

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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

[Joao Lagarto]

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Hi Hanna,
Erythrosin B in water (tau  = 90 ps, refs. Talbot et al (2015) Journal
of Fluorescence & Boens et al (2007) Anal. Chem) should give you a nice IRF.
Cheers,
João

João Lagarto, PhD
Instituto Gulbenkian de Ciência
Rua da Quinta Grande, 6
2780-156 Oeiras
Portugal



Às 12:49 de 19/11/2015, Hanna Sas Nowosielska escreveu:

> *****
> 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
>
>
>

[Kurt Anderson-3]

try gold nano rods, as described here:
Application of ultrafast gold luminescence to measuring the instrument response function for multispectral multiphoton fluorescence lifetime imaging.
Optics Express: 2011 Jul 18;19(15):13848-61. doi: 10.1364/OE.19.013848.




Prof. Kurt I. Anderson
Tumor Cell Migration Lab and
Beatson Advanced Imaging Resource (BAIR)
The Beatson Institute for Cancer Research
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On 19 Nov 2015, at 12:49, Hanna Sas Nowosielska <[hidden email]<mailto:[hidden email]>> wrote:

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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

[Peter Kapusta]

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Erythrosin B's lifetime is still too long, but you can make it shorter by
saturating the solution by KI (Potassium Iodide):
http://dx.doi.org/10.1366/000370209787598979

The same trick works with Rose Bengal as well:
http://www.sciencedirect.com/science/article/pii/S0009261409001389

The dye LDS 798 is another alternative
http://scitation.aip.org/content/aip/journal/rsi/80/3/10.1063/1.3095677

Best regards,
Peter

[yuansheng sun]

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 <[hidden email]>
Date:11/19/2015  6:49 AM  (GMT-06:00)
To: [hidden email]
Cc:  
Subject: Measuring IRF in red channel

*****
To join, leave or search the confocal microscopy listserv, go to:
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*****

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_SN]

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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 <[hidden email]>:

> 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 <[hidden email]>
> Date:11/19/2015  6:49 AM  (GMT-06:00)
> To: [hidden email]
> 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

[Vladimir Ghukasyan-2]

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Dear Hanna,

There may be several reasons to the problem depending on the pattern. It
may be "ringing" or reflections. While the first one can be fixed by
adjusting the threshold of the signal (increasing the baseline and getting
rid of some noise in the microscope environment), the second problem will
be more difficult to address in a turn-key system. This may be coming from
two reflecting surfaces (mirrors or filters). It is most probably in the
laser coupling part of the setup. This pattern will have to be addressed
anyway before you can do any measurements.

Best wishes,
Vladimir


On Fri, Nov 20, 2015 at 3:14 AM, Hanna Sas Nowosielska <
[hidden email]> wrote:

> *****
> 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.
> *****
>
> 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 <[hidden email]>:
>
> > 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 <[hidden email]>
> > Date:11/19/2015  6:49 AM  (GMT-06:00)
> > To: [hidden email]
> > 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
>

[yuansheng sun]

*****
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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 <
[hidden email]> wrote:

> *****
> 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.
> *****
>
> 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 <[hidden email]>:
>
> > 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 <[hidden email]>
> > Date:11/19/2015  6:49 AM  (GMT-06:00)
> > To: [hidden email]
> > 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
>