Third FP options (CFP alternatives?)

[Adrian Smith-6]

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Hi all,

One of our users is looking to set up with a third fluorescent protein for some imaging studies (confocal and multiphoton).

He currently has components labelled with GFP (or YFP in some experiments) and mCherry so he is considering something either in the BFP/CFP range or potentially in the red range past mCherry.

I'm interested in people thoughts on what are the best options these days? There seems to be bewildering array of newer FPs available...

(eg the Evrogen Turbo/Tag reagents like TurboFP650/TagBFP/TagCFP look good but may not work as needs to be able to subclone in order to get into a Leishmania parasite).

Regards,

Adrian Smith
Centenary Institute, Sydney, Australia
 

[Kurt Thorn]

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  We've have good luck with using TagBFP excited by 405nm light in
conjunction with GFP and mCherry.  An improved TagBFP2 is supposed to be
coming soon.  In the far-red channel, some of our users have had good
luck with the recently published IFP1.4, although this requires
biliverdin as a cofactor.  It's readily excited at 640nm.

Best,
Kurt

On 7/5/2011 5:18 AM, Adrian Smith wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
> One of our users is looking to set up with a third fluorescent protein for some imaging studies (confocal and multiphoton).
>
> He currently has components labelled with GFP (or YFP in some experiments) and mCherry so he is considering something either in the BFP/CFP range or potentially in the red range past mCherry.
>
> I'm interested in people thoughts on what are the best options these days? There seems to be bewildering array of newer FPs available...
>
> (eg the Evrogen Turbo/Tag reagents like TurboFP650/TagBFP/TagCFP look good but may not work as needs to be able to subclone in order to get into a Leishmania parasite).
>
> Regards,
>
> Adrian Smith
> Centenary Institute, Sydney, Australia
>
>

[Daniel Gitler]

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Hi,
I have also worked with TagBFP to good effect in neurons with the 405nm laser. It is very well separated from EGFP, and did not show any cross excitation with the 488nm laser.
Daniel


----- Original Message -----
From: Kurt Thorn <[hidden email]>
Date: Tuesday, July 5, 2011 20:07
Subject: Re: Third FP options (CFP alternatives?)
To: [hidden email]

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
>   We've have good luck with using TagBFP excited by 405nm
> light in
> conjunction with GFP and mCherry.  An improved TagBFP2 is
> supposed to be
> coming soon.  In the far-red channel, some of our users
> have had good
> luck with the recently published IFP1.4, although this requires
> biliverdin as a cofactor.  It's readily excited at 640nm.
>
> Best,
> Kurt
>
> On 7/5/2011 5:18 AM, Adrian Smith wrote:
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Hi all,
> >
> > One of our users is looking to set up with a third fluorescent
> protein for some imaging studies (confocal and multiphoton).
> >
> > He currently has components labelled with GFP (or YFP in some
> experiments) and mCherry so he is considering something either
> in the BFP/CFP range or potentially in the red range past mCherry.
> >
> > I'm interested in people thoughts on what are the best options
> these days? There seems to be bewildering array of newer FPs
> available...>
> > (eg the Evrogen Turbo/Tag reagents like
> TurboFP650/TagBFP/TagCFP look good but may not work as needs to
> be able to subclone in order to get into a Leishmania parasite).
> >
> > Regards,
> >
> > Adrian Smith
> > Centenary Institute, Sydney, Australia
> >
> >
>

Daniel Gitler, Ph.D.
Department of Physiology and Neurobiology
Faculty of Health Sciences
Ben Gurion University of the Negev
Beer-Sheva 84105
Israel

Tel:  [phone]
Cell: [phone]
Fax: + 972-8-6477628
http://web2.bgu.ac.il/physiology/faculty-members/daniel-gitler/‎

[Vitaly Boyko]

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I guess CeFP would be also a good choice for CeFP, EYFP and mCherry three
channel imaging applications (440nm, 505/515nm, 568nm sapphire laser lines). It
could useful if someone could provide comparative S/N ratio data for CeFP vs
TagBFP in axons, for example, or any other 'working' environment.
Vitaly




________________________________
From: Daniel Gitler <[hidden email]>
To: [hidden email]
Sent: Tue, July 5, 2011 3:56:35 PM
Subject: Re: Third FP options (CFP alternatives?)

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi,
I have also worked with TagBFP to good effect in neurons with the 405nm laser.
It is very well separated from EGFP, and did not show any cross excitation with
the 488nm laser.
Daniel


----- Original Message -----
From: Kurt Thorn <[hidden email]>
Date: Tuesday, July 5, 2011 20:07
Subject: Re: Third FP options (CFP alternatives?)
To: [hidden email]

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
>   We've have good luck with using TagBFP excited by 405nm
> light in
> conjunction with GFP and mCherry.  An improved TagBFP2 is
> supposed to be
> coming soon.  In the far-red channel, some of our users
> have had good
> luck with the recently published IFP1.4, although this requires
> biliverdin as a cofactor.  It's readily excited at 640nm.
>
> Best,
> Kurt
>
> On 7/5/2011 5:18 AM, Adrian Smith wrote:
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Hi all,
> >
> > One of our users is looking to set up with a third fluorescent
> protein for some imaging studies (confocal and multiphoton).
> >
> > He currently has components labelled with GFP (or YFP in some
> experiments) and mCherry so he is considering something either
> in the BFP/CFP range or potentially in the red range past mCherry.
> >
> > I'm interested in people thoughts on what are the best options
> these days? There seems to be bewildering array of newer FPs
> available...>
> > (eg the Evrogen Turbo/Tag reagents like
> TurboFP650/TagBFP/TagCFP look good but may not work as needs to
> be able to subclone in order to get into a Leishmania parasite).
> >
> > Regards,
> >
> > Adrian Smith
> > Centenary Institute, Sydney, Australia
> >
> >
>

Daniel Gitler, Ph.D.
Department of Physiology and Neurobiology
Faculty of Health Sciences
Ben Gurion University of the Negev
Beer-Sheva 84105
Israel

Tel:  [phone]
Cell: [phone]
Fax: + 972-8-6477628
http://web2.bgu.ac.il/physiology/faculty-members/daniel-gitler/‎