From xpfhd041en at aol.com Mon Dec 8 14:55:33 2003 From: xpfhd041en at aol.com (Ashlee Ziegler) Date: Tue Mar 2 07:54:00 2004 Subject: [FRET] Fw: Credit Declined, application Message-ID: An HTML attachment was scrubbed... URL: http://six.pairlist.net/pipermail/fret/attachments/20031208/e68c1e75/attachment.htm From cgfshm at netscape.net Mon Dec 8 23:58:40 2003 From: cgfshm at netscape.net (Tommie Cole) Date: Tue Mar 2 07:54:00 2004 Subject: [FRET] Declined Credit Application Message-ID: <1-$7$02w60d6az$c1$j@3n643k.knd39> An HTML attachment was scrubbed... URL: http://six.pairlist.net/pipermail/fret/attachments/20031209/26d7c7c3/attachment.htm From fiw at fretimaging.org Tue Dec 9 13:49:35 2003 From: fiw at fretimaging.org (fiw@fretimaging.org) Date: Tue Mar 2 07:54:00 2004 Subject: [FRET] Information from the list owner Message-ID: <1257.69.160.38.22.1070995775.squirrel@webmail.pair.com> Dear list subscribers - I want to start be apologizing for a couple of spam emails that have made their way onto the list. We will in no way tolerate spam and have taken steps to prevent this from happening in the future. The other concern is that there has been no legitimate activity on the list for some time. In my experience, these things require a little push to get started and then take a life of their own. I have been working on a detailed introduction to FRET physics that I plan to post to the site soon. I hope to eventually supplement this with demonstrative Java applets and/or flash animations. While there are actually a significant number of subscribers to the list, there have been almost no postings. To get some dialog started, let me suggest some possible topics for posting to the list: 1. Suggest some content that would be helpful on the fretimaging.org site. 2. Write a brief summary of a recent, interesting paper you have written or read involving FRET. 3. Ask a question about hardware or software currently used by other investigators. If you have any other questions or concerns, please feel free to contact us directly at webmaster@fretimaging.org. Sincerely, Daniel Tyreus, PhD tyreus@socrates.berkeley.edu From GMcnamara at chla.usc.edu Wed Dec 10 17:49:35 2003 From: GMcnamara at chla.usc.edu (Mcnamara, George) Date: Tue Mar 2 07:54:00 2004 Subject: [FRET] FRET review Message-ID: Dear Daniel, Ok, I'll start the FRET reviews off: FICInbt896Elizabeth A Jares-Erijman & Thomas M Jovin (2003) FRET Imaging. Nature Biotechnol 21(11): 1387-1395. Abstract: Forster (or Fluorescence) Resonance Energy Transfer (FRET) is unique in generating fluorescence signals sensitive to molecular conformation, association, and separation in the 1-10 nm range. We introduce a revised photophysical framework for the phenomenon and provide a systematic catalog of FRET techniques adapted to imaging systems, including new approaches proposed as suitable prospects for implementation. Applications extending from a single molecule to live cells will benefit from multidimensional microscopy techniques, particularly those adapted for optical sectioning and incorporating new algorithms for resolving the component contributions to images of complex molecular systems. Review: Essential review on FRET imaging techniques, equations. authors introduce a very nice formalism of treating the flurorophore as a "photon conversion catalyst", i.e. as an 'enzyme' to which Michaelis-Menton kinetics can be applied. Their table 1, taking up most of one page, classifies FRET microscopy methods in categories of Donor quenching and/or acceptor sensitization and of emission anisotropy (each with several subcategories, equations, references). Many are lifetime or anisotropy (fluorescence polarization). They suggest several new approaches. They also define a "Forster Factor" [theta], graph it, and discuss consequences with respect to donor quantum yield. Their Figure 4 shows many fluorophores and protein carriers to the same scale (GFP's, Europium chelate, FlAsH-CCXXCC, Alexa 488, Cy3, Atto 565, QDot 565, strpetavidin and IgG (incidentally, I was told by Chris Baumann of Chroma Tech that the slightly larger QDot 655 is much brighter than the other commercial fluorescent nanocrystals. The E.c. and QY data I recieved from QDot Corp bears this out). Hopefully some company will turn Figure 4 into a freebie wall poster. Comment: Essentially no discussion of "spectral FRET" besides the cryptic subcategory Ia1 (2m3 signals; spectra). Given the number of spectral confocal microscopes on the market (Leica SP2, SP2 AOBS, Zeiss Meta, Bio-Rad's pseudo-spectral filter gizmo), along with Lightform's PARISS (which can be used as a slit scanning confocal), and Applied Spectral Imaging's SD-300 ("SKY") system (which can be used with a spinning disk confocal), and the Jovin's lab expertise in spectral imaging microscopy [search pubmed for: Jovin TM spectral], I was surprised not to see any discussion of "spectral unmixing" approaches to quantifying FRET. Many more of us have 'spectral' confocals than 'lifetime' imagers. Maybe next time? Special note: The November 2003 issue (vol 21, #11) of Nature Biotechnology has as the featured articles an series called Focus on Optical Imaging. The "New on the market" section is on imaging & microscopy. The December issue of NBt had a nice article by Roger Tsien on ReASH CALI/FALI. See also the 23 October issue of Nature (not NBt) for its Imaging Technology Focus article & table of suppliers (pages 867-875) and the Baumgart et al (Watt Webb) multiphoton article, page 821. You may also want to look at the September 2003 issue of Nature Cell Biol / Nature Reviews Mol Cell Biol "Focus on imaging in cell biology (the online version at http://www.nature.com/focus/cellbioimaging/index.html has additional content not in print), and the 4 April 2003 issue of Science "Biological Imaging" ( http://www.sciencemag.org/content/vol300/issue5616/index.shtml) plus the Science magazine's Web Extra" ( http://www.sciencemag.org/feature/data/bioimaging/index.shtml). Recommendation: read the FRET article and look at all the other imaging articles in the 11/2003 issue of NBt. George McNamara, Ph.D. Imaging Scientist Congressman Julian Dixon Image Core The Saban Research Institute of Childrens Hospital Los Angeles 4650 Sunset Blvd., MS 84, SRT 1016 Los Angeles, CA 90027 323-669-2548 voice 323-251-8878 cell 323-668-7921 fax gmcnamara@chla.usc.edu http://shelia/image Image Core Web (CHLA Intranet) \\ntapps49\home Image Core User Home (Intranet) \\ntapps49\appnotes Image Core Application Notes (Intranet) http://www.childrenshospitalla.org/research.html CHLA Research Institute < http://home.earthlink.net/~geomcnamara > Personal web site -----Original Message----- From: fret-request@fretimaging.org [ mailto:fret-request@fretimaging.org ] Sent: Wednesday, December 10, 2003 1:21 AM To: fret@fretimaging.org Subject: FRET digest, Vol 1 #6 - 1 msg Send FRET mailing list submissions to fret@fretimaging.org To subscribe or unsubscribe via the World Wide Web, visit http://four.pairlist.net/mailman/listinfo/fret or, via email, send a message with subject or body 'help' to fret-request@fretimaging.org You can reach the person managing the list at fret-admin@fretimaging.org When replying, please edit your Subject line so it is more specific than "Re: Contents of FRET digest..." Today's Topics: 1. Information from the list owner (fiw@fretimaging.org) --__--__-- Message: 1 Date: Tue, 9 Dec 2003 13:49:35 -0500 (EST) From: To: Subject: [FRET] Information from the list owner Dear list subscribers - I want to start be apologizing for a couple of spam emails that have made their way onto the list. We will in no way tolerate spam and have taken steps to prevent this from happening in the future. The other concern is that there has been no legitimate activity on the list for some time. In my experience, these things require a little push to get started and then take a life of their own. I have been working on a detailed introduction to FRET physics that I plan to post to the site soon. I hope to eventually supplement this with demonstrative Java applets and/or flash animations. While there are actually a significant number of subscribers to the list, there have been almost no postings. To get some dialog started, let me suggest some possible topics for posting to the list: 1. Suggest some content that would be helpful on the fretimaging.org site. 2. Write a brief summary of a recent, interesting paper you have written or read involving FRET. 3. Ask a question about hardware or software currently used by other investigators. If you have any other questions or concerns, please feel free to contact us directly at webmaster@fretimaging.org. Sincerely, Daniel Tyreus, PhD tyreus@socrates.berkeley.edu --__--__-- _______________________________________________ FRET mailing list FRET@fretimaging.org http://four.pairlist.net/mailman/listinfo/fret End of FRET Digest -------------- next part -------------- An HTML attachment was scrubbed... URL: http://six.pairlist.net/pipermail/fret/attachments/20031210/1162f243/attachment.htm