CONFOCALMICROSCOPY Archives

July 2013

CONFOCALMICROSCOPY@LISTS.UMN.EDU
Options: Use Monospaced Font
Show Text Part by Default
Show All Mail Headers

Message: [<< First] [< Prev] [Next >] [Last >>]
Topic: [<< First] [< Prev] [Next >] [Last >>]
Author: [<< First] [< Prev] [Next >] [Last >>]

Print Reply
Subject:
Re: Red fluorescent proteins suitable for two-color two-photon microscopy and protein tagging
From:
Tobias Rose <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Tue, 2 Jul 2013 17:00:21 +0000
Content-Type:
text/plain
Parts/Attachments:
text/plain (29 lines) 
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

This is indeed a bit on the short side. However, in your test this should hurt mCherry even more than mRuby2; tdtomato should be favored  (emission: mCherry 610 vs mRuby2 600 vs. tdtomato 580). 

I use this filter:
http://www.semrock.com/FilterDetails.aspx?id=FF01-607/70-25

Tobias



-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Steven Henle
Sent: Tuesday, July 02, 2013 18:10
To: [log in to unmask]
Subject: Re: Red fluorescent proteins suitable for two-color two-photon microscopy and protein tagging

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

I did use the same mRuby2 construct from addgene, and like you started and the normal GFP start site. I just rechecked the sequence too and everything looks perfect. I am setting up to test it again on the 2-photon. Part of the issue may be the filter I am using (565-610), which is probably not far enough red to be optimal, but should catch over half of the signal. What filter do you use?

Steve

ATOM RSS1 RSS2