Locate some easy-to-use long lifetime fluorphores?

[Tom Lawson]

Dear Confocalists,

I am trying to locate some commercial and easy-to-use long lifetime
fluorphores to time-gate autofluorescence. Ideally they would have to
last at least 1ms (which is a long time).

The research team I work with makes its own lanthanides organometallic
fluorophores Eu3+ and Tb3+ which last about 3ms. But I am anticipating
getting specimens from collaborators who may want to label the
specimens at their end and lanthanides are not easy to use.

I thought I might be able to find some organometallic transition
metal-ligand complexes such as ruthenium (Ru II), rhenium (Re I), or
osmium (Os II) (10 ns to 10 μs) or some organic fluorophores such as
Pyrene (400ns) or Coronenes (200ns). But I have had no luck finding
any, they might not be easy to use and 400ns is cutting it a bit
short.

One suggestion I have had so far is quantum dots which I need to look
into. There maybe other options I am not aware of.

I would be grateful if you have any suggestions?

Regards,

--
Tom Lawson
[hidden email]
PhD Student,
Macquarie University
NSW, 2109
Australia

[Morrison, Ian E]

Tom, you might like to look at triplet probes such as eosin or erythrosin.  Eosin was used extensively in the 1980s for rotational diffusion measurements (keywords: transient dichroism) and can have lifetimes up to 2ms but is sensitive to quenching.  The eosin is introduced into proteins via the maleimide or isothiocyanate reactive groups, both from Invitrogen (no commercial interest).  Quenching was prevented by rigorous exclusion of oxygen, or it can also be a source of information about the accessibility of the eosin (exposed/buried).  Erythrosin was used for phosphorescence studies (check publications from Bill Sawyer at Melbourne University).  I have used both probes in widefield imaging, but not often by confocal as they have relatively low intensity and the long lifetime gave problems at normal scan rates.

If you need more information, contact me offline at my new address ([hidden email]).
Best wishes, Ian

Dr. I.E.G. Morrison, Biology Department, University of York, York UK.

-----Original Message-----
From: Confocal Microscopy List on behalf of Tom Lawson
Sent: Sat 18/04/2009 07:08
To: [hidden email]
Subject: Locate some easy-to-use long lifetime fluorphores?
 
Dear Confocalists,

I am trying to locate some commercial and easy-to-use long lifetime
fluorphores to time-gate autofluorescence. Ideally they would have to
last at least 1ms (which is a long time).

The research team I work with makes its own lanthanides organometallic
fluorophores Eu3+ and Tb3+ which last about 3ms. But I am anticipating
getting specimens from collaborators who may want to label the
specimens at their end and lanthanides are not easy to use.

I thought I might be able to find some organometallic transition
metal-ligand complexes such as ruthenium (Ru II), rhenium (Re I), or
osmium (Os II) (10 ns to 10 ?s) or some organic fluorophores such as
Pyrene (400ns) or Coronenes (200ns). But I have had no luck finding
any, they might not be easy to use and 400ns is cutting it a bit
short.

One suggestion I have had so far is quantum dots which I need to look
into. There maybe other options I am not aware of.

I would be grateful if you have any suggestions?

Regards,

--
Tom Lawson
[hidden email]
PhD Student,
Macquarie University
NSW, 2109
Australia

[Sonja Hatz]

Tom,

Although I have no personal experience, in our lab Rhuthenium dyes  
have been used. We also gate the fluorescence and find we can use  
these compounds with lifetimes around 500-800ns. We used them for  
oxygen sensing in cell cultures, so there are protocols to introduce  
them into cells and dark toxicity studies.

e.g. Ru(bipy), RuDPP and Ru(phen). Several compounds are available from Sigma

Sonja

Citat af Tom Lawson <[hidden email]>:

> Dear Confocalists,
>
> I am trying to locate some commercial and easy-to-use long lifetime
> fluorphores to time-gate autofluorescence. Ideally they would have to
> last at least 1ms (which is a long time).
>
> The research team I work with makes its own lanthanides organometallic
> fluorophores Eu3+ and Tb3+ which last about 3ms. But I am anticipating
> getting specimens from collaborators who may want to label the
> specimens at their end and lanthanides are not easy to use.
>
> I thought I might be able to find some organometallic transition
> metal-ligand complexes such as ruthenium (Ru II), rhenium (Re I), or
> osmium (Os II) (10 ns to 10 μs) or some organic fluorophores such as
> Pyrene (400ns) or Coronenes (200ns). But I have had no luck finding
> any, they might not be easy to use and 400ns is cutting it a bit
> short.
>
> One suggestion I have had so far is quantum dots which I need to look
> into. There maybe other options I am not aware of.
>
> I would be grateful if you have any suggestions?
>
> Regards,
>
> --
> Tom Lawson
> [hidden email]
> PhD Student,
> Macquarie University
> NSW, 2109
> Australia
>

[Jurriaan Zwier]

Commercial response:

Dear Tom,

We are specialised in luminescent lanthanide probes for drug discovery
applications. We have an europium cryptate NHS or maleimide available with
a lifetime of about 1 ms in biological media. FRET studies can be
performed easily, we use this in our assays. They are very stable wich is
a great advantage over other lanthanide chelates available. Antibody or
protein-labeling in general is not problematic.
Evidently they can be used for other biological purpuses as well,
including microscopy, but this depends strongly on your budget :-)
If you want more info you can have a look at www.htrf.com or contact our
service department. You can also contact me personally at
[hidden email] (I use an old email address on this list)
For the best (in my humble opinion) terbium based probe you can have a
look at www.lumiphore.com
For a recent paper on FRET studies with this kind of probes for membrane
receptor studies see:
D. Maurel et al, Nature Methods 5 (2008) p561-567

Hope this gives some leads,
Kind regards,

Jurriaan Zwier, Ph.D, research scientist at cisbio-bioassays.

> Dear Confocalists,
>
> I am trying to locate some commercial and easy-to-use long lifetime
> fluorphores to time-gate autofluorescence. Ideally they would have to
> last at least 1ms (which is a long time).
>
> The research team I work with makes its own lanthanides organometallic
> fluorophores Eu3+ and Tb3+ which last about 3ms. But I am anticipating
> getting specimens from collaborators who may want to label the
> specimens at their end and lanthanides are not easy to use.
>
> I thought I might be able to find some organometallic transition
> metal-ligand complexes such as ruthenium (Ru II), rhenium (Re I), or
> osmium (Os II) (10 ns to 10 ìs) or some organic fluorophores such as
> Pyrene (400ns) or Coronenes (200ns). But I have had no luck finding
> any, they might not be easy to use and 400ns is cutting it a bit
> short.
>
> One suggestion I have had so far is quantum dots which I need to look
> into. There maybe other options I am not aware of.
>
> I would be grateful if you have any suggestions?
>
> Regards,
>
> --
> Tom Lawson
> [hidden email]
> PhD Student,
> Macquarie University
> NSW, 2109
> Australia
>