Fwd: Best far red fluorescent protein?

[Bojana]

*****
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.
*****

Microscopy geeks,

What’s your preferred far-red fluorescent protein, brighter/more stable
than mcherry? Mplum, mkate, mcardinal, fusion red?

We want to use it as Lifeact tag to image cytoskeleton over 2-4 days, in
addition to blue, green and red proteins.

Bojana

[Heddleston, John]

*****
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 Bojana,
I really like to use the HALO tag system with the Janelia Fluor ligands. JF646 works very well...

If you can't use HALO, iRFP-670 has worked well. If you need more in the "middle ground" (590nm), I like mKate2.

Cheers,
John

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Bojana Gligorijevic
Sent: Friday, February 8, 2019 1:14 PM
To: [hidden email]
Subject: Fwd: Best far red fluorescent protein?

*****
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.
*****

Microscopy geeks,

What’s your preferred far-red fluorescent protein, brighter/more stable than mcherry? Mplum, mkate, mcardinal, fusion red?

We want to use it as Lifeact tag to image cytoskeleton over 2-4 days, in addition to blue, green and red proteins.

Bojana

[Philipp Tripal]

*****
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 Bojana


we have made good experiences with mScarlet. Each variant of mScarlet should enable the imaging of the actin cytoskeleton for 2-4 days. It´s not really far red but super bright, monomeric and highly stable in living cells.


Best regards

Philipp




Philipp Tripal
Optical Imaging Centre Erlangen

Hartmannstr. 14
91052 Erlangen, Germany
[phone] (Office)
[phone] (Secretary)
[phone] (Fax)

www.oice.uni-erlangen.de<https://groupware.fau.de/owa/redir.aspx?SURL=qGsZqxfKB1wge1J3khFcYM_9PCMB9VodNi5E85QS6SrIhCN-tOXSCGgAdAB0AHAAOgAvAC8AdwB3AHcALgBvAGkAYwBlAC4AdQBuAGkALQBlAHIAbABhAG4AZwBlAG4ALgBkAGUA&URL=http%3a%2f%2fwww.oice.uni-erlangen.de>
________________________________
Von: Confocal Microscopy List <[hidden email]> im Auftrag von Bojana Gligorijevic <[hidden email]>
Gesendet: Freitag, 8. Februar 2019 19:13:40
An: [hidden email]
Betreff: Fwd: Best far red fluorescent protein?

*****
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.
*****

Microscopy geeks,

What’s your preferred far-red fluorescent protein, brighter/more stable
than mcherry? Mplum, mkate, mcardinal, fusion red?

We want to use it as Lifeact tag to image cytoskeleton over 2-4 days, in
addition to blue, green and red proteins.

Bojana

*****
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 Bojana,

I agree with John. If you can get away with using a HALO- or SNAP-tag coupled to an organic, membrane permeable fluorophore, you will have the best results concerning photostability and brightness. I can recommend SiR (silicone rhodamine) for this. Far-red, bright and fluorogenic. You can purchase the SiR-SNAP construct, but the SiR-HALO you would need to couple yourself.

As for actual far-red fluorescent proteins, we have been well-served by mNeptune2, mNeptune2.5 or mCardinal, which were all very similar. mKate2 is also a good choice. But none come close to the excellence of organic dyes (again: if you can get way with them).

Good luck!
Nicolai

-----------------------------------------------------------------
Max Planck Florida Institute for Neuroscience
1 Max Planck Way
Jupiter, FL 33458

________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Heddleston, John <[hidden email]>
Sent: Friday, February 8, 2019 1:34 PM
To: [hidden email]
Subject: Re: Best far red fluorescent protein?

*****
To join, leave or search the confocal microscopy listserv, go to:
https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=02%7C01%7CNicolai.Urban%40MPFI.ORG%7C4da230a1c73c483683c908d68e9bb016%7C947b45517db44636a5fd1bdcad603ed0%7C0%7C0%7C636853196667607727&amp;sdata=ZyQy8QkUutQrfPvahz7h5wguBfJcOkftWtJzS1mE8jo%3D&amp;reserved=0
Post images on https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&amp;data=02%7C01%7CNicolai.Urban%40MPFI.ORG%7C4da230a1c73c483683c908d68e9bb016%7C947b45517db44636a5fd1bdcad603ed0%7C0%7C0%7C636853196667607727&amp;sdata=9oKmCzVGkjZKF2W4%2F4iq2sF0ESzVAImrPitzQ%2B9JG50%3D&amp;reserved=0 and include the link in your posting.
*****

Hi Bojana,
I really like to use the HALO tag system with the Janelia Fluor ligands. JF646 works very well...

If you can't use HALO, iRFP-670 has worked well. If you need more in the "middle ground" (590nm), I like mKate2.

Cheers,
John

-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Bojana Gligorijevic
Sent: Friday, February 8, 2019 1:14 PM
To: [hidden email]
Subject: Fwd: Best far red fluorescent protein?

*****
To join, leave or search the confocal microscopy listserv, go to:
https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&amp;data=02%7C01%7CNicolai.Urban%40MPFI.ORG%7C4da230a1c73c483683c908d68e9bb016%7C947b45517db44636a5fd1bdcad603ed0%7C0%7C0%7C636853196667607727&amp;sdata=ZyQy8QkUutQrfPvahz7h5wguBfJcOkftWtJzS1mE8jo%3D&amp;reserved=0
Post images on https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&amp;data=02%7C01%7CNicolai.Urban%40MPFI.ORG%7C4da230a1c73c483683c908d68e9bb016%7C947b45517db44636a5fd1bdcad603ed0%7C0%7C0%7C636853196667607727&amp;sdata=9oKmCzVGkjZKF2W4%2F4iq2sF0ESzVAImrPitzQ%2B9JG50%3D&amp;reserved=0 and include the link in your posting.
*****

Microscopy geeks,

What’s your preferred far-red fluorescent protein, brighter/more stable than mcherry? Mplum, mkate, mcardinal, fusion red?

We want to use it as Lifeact tag to image cytoskeleton over 2-4 days, in addition to blue, green and red proteins.

Bojana