Rhodamine X (ROX)

[Cameron Nowell-3]

*****
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.
*****


Hi List

Has anyone had any experience with Rhodamine X? It seems like it should be amazingly bright and fairly useful. Is it fairly photo stable? And has anyone tried it in STORM imaging?

We are about to get a heap of peptides made up for various things and wondering what would be the best tag to have put on them.

Cheers
Cam



Cameron J. Nowell
Research Facilities Manager

Imaging, FACS and Analysis Core
Monash Institute of Pharmaceutical Sciences
Monash University
399 Royal Parade
(Mail address: 381 Royal Parade)
Parkville, VIC, 3052
Australia

Email: [hidden email]<mailto:[hidden email]>
Mobile: [phone]
Office: [phone]

LinkedIn: Profile<http://au.linkedin.com/pub/cameron-nowell/23/57/884/>
Research Gate:  Profile<http://www.researchgate.net/profile/Cameron_Nowell>

[Aleksander Szczurek]

*****
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.
*****

Hello Cameron,
I think it may be pretty promising. Check following paper by Carlini and
Manley:
http://www.ncbi.nlm.nih.gov/pubmed/24079385
They optimized imaging conditions for localization microscopy of Rhodamine
123.

greetings from Mainz
Aleksander


2014-08-14 8:16 GMT+02:00 Cameron Nowell <[hidden email]>:

> *****
> 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.
> *****
>
>
> Hi List
>
> Has anyone had any experience with Rhodamine X? It seems like it should be
> amazingly bright and fairly useful. Is it fairly photo stable? And has
> anyone tried it in STORM imaging?
>
> We are about to get a heap of peptides made up for various things and
> wondering what would be the best tag to have put on them.
>
> Cheers
> Cam
>
>
>
> Cameron J. Nowell
> Research Facilities Manager
>
> Imaging, FACS and Analysis Core
> Monash Institute of Pharmaceutical Sciences
> Monash University
> 399 Royal Parade
> (Mail address: 381 Royal Parade)
> Parkville, VIC, 3052
> Australia
>
> Email: [hidden email]<mailto:[hidden email]>
> Mobile: [phone]
> Office: [phone]
>
> LinkedIn: Profile<http://au.linkedin.com/pub/cameron-nowell/23/57/884/>
> Research Gate:  Profile<http://www.researchgate.net/profile/Cameron_Nowell
> >
>

[Iain Johnson]

*****
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.
*****

Hi Cam:

ROX is indeed a very photostable dye.  Its main drawback is that it is highly non polar (greasy). If you are singly labeling peptides, this may not be so much of a problem as it tends to be when multiple labeling an antibody. If you want something more polar but spectroscopically near-equivalent, that would be AF568.

Iain

Sent from my iPhone

On Aug 13, 2014, at 11:16 PM, Cameron Nowell <[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.
> *****
>
>
> Hi List
>
> Has anyone had any experience with Rhodamine X? It seems like it should be amazingly bright and fairly useful. Is it fairly photo stable? And has anyone tried it in STORM imaging?
>
> We are about to get a heap of peptides made up for various things and wondering what would be the best tag to have put on them.
>
> Cheers
> Cam
>
>
>
> Cameron J. Nowell
> Research Facilities Manager
>
> Imaging, FACS and Analysis Core
> Monash Institute of Pharmaceutical Sciences
> Monash University
> 399 Royal Parade
> (Mail address: 381 Royal Parade)
> Parkville, VIC, 3052
> Australia
>
> Email: [hidden email]<mailto:[hidden email]>
> Mobile: [phone]
> Office: [phone]
>
> LinkedIn: Profile<http://au.linkedin.com/pub/cameron-nowell/23/57/884/>
> Research Gate:  Profile<http://www.researchgate.net/profile/Cameron_Nowell>

[Cameron Nowell-3]

*****
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.
*****

Hi Ian,

Awesome. We mainly want to label ligands with single tags to do some live cell binding and TIRF so sounds like it would be great. AF568 is an option but the cost of ROX per mg is much much lower.

Cheers

Cam


Sent from Samsung Mobile


-------- Original message --------
From: Iain Johnson
Date:15/08/2014 1:05 AM (GMT+10:00)
To: [hidden email]
Subject: Re: Rhodamine X (ROX)

*****
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.
*****

Hi Cam:

ROX is indeed a very photostable dye.  Its main drawback is that it is highly non polar (greasy). If you are singly labeling peptides, this may not be so much of a problem as it tends to be when multiple labeling an antibody. If you want something more polar but spectroscopically near-equivalent, that would be AF568.

Iain

Sent from my iPhone

On Aug 13, 2014, at 11:16 PM, Cameron Nowell <[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.
> *****
>
>
> Hi List
>
> Has anyone had any experience with Rhodamine X? It seems like it should be amazingly bright and fairly useful. Is it fairly photo stable? And has anyone tried it in STORM imaging?
>
> We are about to get a heap of peptides made up for various things and wondering what would be the best tag to have put on them.
>
> Cheers
> Cam
>
>
>
> Cameron J. Nowell
> Research Facilities Manager
>
> Imaging, FACS and Analysis Core
> Monash Institute of Pharmaceutical Sciences
> Monash University
> 399 Royal Parade
> (Mail address: 381 Royal Parade)
> Parkville, VIC, 3052
> Australia
>
> Email: [hidden email]<mailto:[hidden email]>
> Mobile: [phone]
> Office: [phone]
>
> LinkedIn: Profile<http://au.linkedin.com/pub/cameron-nowell/23/57/884/>
> Research Gate:  Profile<http://www.researchgate.net/profile/Cameron_Nowell>

[Iain Johnson]

*****
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.
*****

Cam:

Couple of other things to bear in mind.  It is probably worth splashing a
bit of extra money to buy the single isomer form of the dye (i.e. 5-ROX or
6-ROX, not 5-(and 6)-ROX.  With the mixed isomers, you are making a mixture
of two products with different binding affinities, even if there's only a
single labeling site.  AF568 is only available as a mixed isomers
preparation.

One of the things that can be anticipated is nonspecific binding of the
labeled peptide mediated by the dye.  One thing that can be done to get an
assessment of this is to use the free dye (i.e. 5-carboxy ROX or
6-carboxy-ROX, NOT the reactive NHS ester forms used to do the peptide
labeling) at the same concentration, incubation time and wash stringency as
the labeled peptide. This is one of those images that you want to be as dim
as possible!.

Iain


On Thu, Aug 14, 2014 at 2:28 PM, Cameron Nowell <[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.
> *****
>
> Hi Ian,
>
> Awesome. We mainly want to label ligands with single tags to do some live
> cell binding and TIRF so sounds like it would be great. AF568 is an option
> but the cost of ROX per mg is much much lower.
>
> Cheers
>
> Cam
>
>
> Sent from Samsung Mobile
>
>
> -------- Original message --------
> From: Iain Johnson
> Date:15/08/2014 1:05 AM (GMT+10:00)
> To: [hidden email]
> Subject: Re: Rhodamine X (ROX)
>
> *****
> 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.
> *****
>
> Hi Cam:
>
> ROX is indeed a very photostable dye.  Its main drawback is that it is
> highly non polar (greasy). If you are singly labeling peptides, this may
> not be so much of a problem as it tends to be when multiple labeling an
> antibody. If you want something more polar but spectroscopically
> near-equivalent, that would be AF568.
>
> Iain
>
> Sent from my iPhone
>
> On Aug 13, 2014, at 11:16 PM, Cameron Nowell <[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.
> > *****
> >
> >
> > Hi List
> >
> > Has anyone had any experience with Rhodamine X? It seems like it should
> be amazingly bright and fairly useful. Is it fairly photo stable? And has
> anyone tried it in STORM imaging?
> >
> > We are about to get a heap of peptides made up for various things and
> wondering what would be the best tag to have put on them.
> >
> > Cheers
> > Cam
> >
> >
> >
> > Cameron J. Nowell
> > Research Facilities Manager
> >
> > Imaging, FACS and Analysis Core
> > Monash Institute of Pharmaceutical Sciences
> > Monash University
> > 399 Royal Parade
> > (Mail address: 381 Royal Parade)
> > Parkville, VIC, 3052
> > Australia
> >
> > Email: [hidden email]<mailto:[hidden email]>
> > Mobile: [phone]
> > Office: [phone]
> >
> > LinkedIn: Profile<http://au.linkedin.com/pub/cameron-nowell/23/57/884/>
> > Research Gate:  Profile<
> http://www.researchgate.net/profile/Cameron_Nowell>
>