Deconvolution of Confocal Images? (was: Airy Units)
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Are you sure there is a 514nm DPSS? I think that wavelength would have to be an OPS and I don't know anyone who makes one. Coherent do make a 505nm OPS which is intended to stand in for both the 488 & 514 lines of an Argon laser, and I rather suspect that this could be exactly what you need. But I have no experience of it. (I do have a Coherent 488nm OPS). Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Australian Centre for Microscopy & Microanalysis, Madsen Building F09, University of Sydney, NSW 2006 Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Tim Feinstein Sent: Thursday, 10 November 2011 7:34 AM To: [hidden email] Subject: FRET in the time of DPSS ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hello all, We want to spec a four-laser launch for a new live cell system that will handle both CFP/YFP FRET and red/green imaging. However, I am sad to see that gas lasers are no longer speccable and so the freebie 514 laser line is gone. We would therefore have to spec a 514 DPSS and forego the far-red line. I was wondering whether there is a way to do more (or at least the same) with less. 488 nm excites YFP well enough, so in theory I could image CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm laser lines. In my experience 442 nm laser excitation (via TIRF) causes negligible YFP excitation and 488 nm does not excite CFP, so it is possible that I could gain speed by passing everything through a single broad bandpass filter (e.g., 455-550 nm) and alternate excitations. Assuming that cross-talk is not a problem, the most significant cost would be that I lose a decent chunk of CFP emission to the scan head dichroic, but in return I gain a 641 nm laser. Has anyone tried this? Any feedback on or off-list would be much appreciated. Thanks and all the best, TF Timothy Feinstein, PhD Postdoctoral Fellow Laboratory for GPCR Biology Dept. of Pharmacology & Chemical Biology University of Pittsburgh, School of Medicine BST W1301, 200 Lothrop St. Pittsburgh, PA 15261 ----- No virus found in this message. Checked by AVG - www.avg.com Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: 11/08/11 |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** See: http://www.cobolt.se/coboltfandango515nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw No connection, just considering about switching away from our old Ar Ion as well. - Damir On 11/9/2011 3:36 PM, Guy Cox wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Are you sure there is a 514nm DPSS? I think that wavelength would have > to be an OPS and I don't know anyone who makes one. Coherent do make a > 505nm OPS which is intended to stand in for both the 488& 514 lines of > an Argon laser, and I rather suspect that this could be exactly what you > need. But I have no experience of it. (I do have a Coherent 488nm > OPS). > > Guy > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor& Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) > Australian Centre for Microscopy& Microanalysis, > Madsen Building F09, University of Sydney, NSW 2006 > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Tim Feinstein > Sent: Thursday, 10 November 2011 7:34 AM > To: [hidden email] > Subject: FRET in the time of DPSS > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello all, > > We want to spec a four-laser launch for a new live cell system that will > handle both CFP/YFP FRET and red/green imaging. However, I am sad to > see that gas lasers are no longer speccable and so the freebie 514 laser > line is gone. We would therefore have to spec a 514 DPSS and forego the > far-red line. > > I was wondering whether there is a way to do more (or at least the same) > with less. 488 nm excites YFP well enough, so in theory I could image > CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm laser > lines. In my experience 442 nm laser excitation (via TIRF) causes > negligible YFP excitation and 488 nm does not excite CFP, so it is > possible that I could gain speed by passing everything through a single > broad bandpass filter (e.g., 455-550 nm) and alternate excitations. > Assuming that cross-talk is not a problem, the most significant cost > would be that I lose a decent chunk of CFP emission to the scan head > dichroic, but in return I gain a 641 nm laser. > > Has anyone tried this? Any feedback on or off-list would be much > appreciated. > > Thanks and all the best, > > > TF > > Timothy Feinstein, PhD > Postdoctoral Fellow > Laboratory for GPCR Biology > Dept. of Pharmacology& Chemical Biology > University of Pittsburgh, School of Medicine > BST W1301, 200 Lothrop St. > Pittsburgh, PA 15261 > > ----- > No virus found in this message. > Checked by AVG - www.avg.com > Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: 11/08/11 -- Damir Sudar - Staff Scientist and Deputy for Technology Lawrence Berkeley Laboratory / Life Sciences Division One Cyclotron Road, MS 977, Berkeley, CA 94720, USA T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] WWW: http://www.lbl.gov/lsd/People_&_Organization/Scientific_Staff_Directory/Sudar_Lab.html |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Industry Response: There absolutely is a 514 DPSS available from 20 mW up to 150 mW available in the Sapphire form factor from Coherent. We use this laser, often in conjunction with the 445 CUBE in our LaserStack for CFP/YFP (and their newer brothers and sisters in FP evolution) imaging for FRET or otherwise. ------------------------ Samuel A. Connell Sales Manager Pacific Region-North America Intelligent Imaging Innovations, Inc 3250 Ocean Park Blvd, Suite 202 Santa Monica, CA 90405 Cell: (858) 692-4510 [hidden email] On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> > ***** > > See: http://www.cobolt.se/**coboltfandango515nm.html?** > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw<http://www.cobolt.se/coboltfandango515nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> > > No connection, just considering about switching away from our old Ar Ion > as well. > > - Damir > > > On 11/9/2011 3:36 PM, Guy Cox wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> >> ***** >> >> Are you sure there is a 514nm DPSS? I think that wavelength would have >> to be an OPS and I don't know anyone who makes one. Coherent do make a >> 505nm OPS which is intended to stand in for both the 488& 514 lines of >> >> an Argon laser, and I rather suspect that this could be exactly what you >> need. But I have no experience of it. (I do have a Coherent 488nm >> OPS). >> >> Guy >> >> Optical Imaging Techniques in Cell Biology >> by Guy Cox CRC Press / Taylor& Francis >> >> http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> >> ______________________________**________________ >> Associate Professor Guy Cox, MA, DPhil(Oxon) >> Australian Centre for Microscopy& Microanalysis, >> >> Madsen Building F09, University of Sydney, NSW 2006 >> >> Phone +61 2 9351 3176 Fax +61 2 9351 7682 >> Mobile 0413 281 861 >> ______________________________**________________ >> http://www.guycox.net >> >> >> >> -----Original Message----- >> From: Confocal Microscopy List [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU<[hidden email]> >> ] >> On Behalf Of Tim Feinstein >> Sent: Thursday, 10 November 2011 7:34 AM >> To: [hidden email].**EDU <[hidden email]> >> Subject: FRET in the time of DPSS >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> >> ***** >> >> Hello all, >> >> We want to spec a four-laser launch for a new live cell system that will >> handle both CFP/YFP FRET and red/green imaging. However, I am sad to >> see that gas lasers are no longer speccable and so the freebie 514 laser >> line is gone. We would therefore have to spec a 514 DPSS and forego the >> far-red line. >> >> I was wondering whether there is a way to do more (or at least the same) >> with less. 488 nm excites YFP well enough, so in theory I could image >> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm laser >> lines. In my experience 442 nm laser excitation (via TIRF) causes >> negligible YFP excitation and 488 nm does not excite CFP, so it is >> possible that I could gain speed by passing everything through a single >> broad bandpass filter (e.g., 455-550 nm) and alternate excitations. >> Assuming that cross-talk is not a problem, the most significant cost >> would be that I lose a decent chunk of CFP emission to the scan head >> dichroic, but in return I gain a 641 nm laser. >> >> Has anyone tried this? Any feedback on or off-list would be much >> appreciated. >> >> Thanks and all the best, >> >> >> TF >> >> Timothy Feinstein, PhD >> Postdoctoral Fellow >> Laboratory for GPCR Biology >> Dept. of Pharmacology& Chemical Biology >> >> University of Pittsburgh, School of Medicine >> BST W1301, 200 Lothrop St. >> Pittsburgh, PA 15261 >> >> ----- >> No virus found in this message. >> Checked by AVG - www.avg.com >> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: 11/08/11 >> > > -- > Damir Sudar - Staff Scientist and Deputy for Technology > Lawrence Berkeley Laboratory / Life Sciences Division > One Cyclotron Road, MS 977, Berkeley, CA 94720, USA > T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] > WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** > Staff_Directory/Sudar_Lab.html<http://www.lbl.gov/lsd/People_&_Organization/Scientific_Staff_Directory/Sudar_Lab.html> > |
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In reply to this post by Tim Feinstein-2
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Ditch the CFP/YFP constructs and go with EGFP (488) and whatever Red (561) FP is working well. That leaves 405 and 633 nm for the other wavelengths into the launch. p.s. no offense to those who have sent me Teal, Cerulean3 and other Cyan containing FRET constructs. On 11/9/2011 3:33 PM, Tim Feinstein wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello all, > > We want to spec a four-laser launch for a new live cell system that will handle both CFP/YFP FRET and red/green imaging. However, I am sad to see that gas lasers are no longer speccable and so the freebie 514 laser line is gone. We would therefore have to spec a 514 DPSS and forego the far-red line. > > I was wondering whether there is a way to do more (or at least the same) with less. 488 nm excites YFP well enough, so in theory I could image CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm laser lines. In my experience 442 nm laser excitation (via TIRF) causes negligible YFP excitation and 488 nm does not excite CFP, so it is possible that I could gain speed by passing everything through a single broad bandpass filter (e.g., 455-550 nm) and alternate excitations. Assuming that cross-talk is not a problem, the most significant cost would be that I lose a decent chunk of CFP emission to the scan head dichroic, but in return I gain a 641 nm laser. > > Has anyone tried this? Any feedback on or off-list would be much appreciated. > > Thanks and all the best, > > > TF > > Timothy Feinstein, PhD > Postdoctoral Fellow > Laboratory for GPCR Biology > Dept. of Pharmacology& Chemical Biology > University of Pittsburgh, School of Medicine > BST W1301, 200 Lothrop St. > Pittsburgh, PA 15261 > > -- George McNamara, PhD Analytical Imaging Core Facility University of Miami |
 
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Zac Arrac Atelaz |
Re: FRET in the time of DPSS
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In reply to this post by Tim Feinstein-2
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Timothy: If you are planning a confocal you can order 405, 440, 559, 635, this combination would do what you need, if you can go for 514dpss, then the money might be enough to have also the 475. This would have all the range in it, and you will have no significant change in the general price of the system. I hope this helps Gabriel OH > Date: Wed, 9 Nov 2011 15:33:30 -0500 > From: [hidden email] > Subject: FRET in the time of DPSS > To: [hidden email] > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hello all, > > We want to spec a four-laser launch for a new live cell system that will handle both CFP/YFP FRET and red/green imaging. However, I am sad to see that gas lasers are no longer speccable and so the freebie 514 laser line is gone. We would therefore have to spec a 514 DPSS and forego the far-red line. > > I was wondering whether there is a way to do more (or at least the same) with less. 488 nm excites YFP well enough, so in theory I could image CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm laser lines. In my experience 442 nm laser excitation (via TIRF) causes negligible YFP excitation and 488 nm does not excite CFP, so it is possible that I could gain speed by passing everything through a single broad bandpass filter (e.g., 455-550 nm) and alternate excitations. Assuming that cross-talk is not a problem, the most significant cost would be that I lose a decent chunk of CFP emission to the scan head dichroic, but in return I gain a 641 nm laser. > > Has anyone tried this? Any feedback on or off-list would be much appreciated. > > Thanks and all the best, > > > TF > > Timothy Feinstein, PhD > Postdoctoral Fellow > Laboratory for GPCR Biology > Dept. of Pharmacology & Chemical Biology > University of Pittsburgh, School of Medicine > BST W1301, 200 Lothrop St. > Pittsburgh, PA 15261 |
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In reply to this post by samuel connell
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** The Sapphire series lasers are OPS (Optically Pumped Semiconductor) not DPSS but, you are right, they do indeed have a 514nm version. I don't know why I didn't find it, but did find the 505, when I searched this morning. I still think that the 505 might be exactly what Timothy needs, though, since he doesn't want to buy an extra laser. I have also been contacted off-list and told that Spectra-Physics have a 515nm DPSS which is close enough to make no difference. I've no idea what the lasing crystal is. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Australian Centre for Microscopy & Microanalysis, Madsen Building F09, University of Sydney, NSW 2006 Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of samuel connell Sent: Thursday, 10 November 2011 1:57 PM To: [hidden email] Subject: Re: FRET in the time of DPSS ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Industry Response: There absolutely is a 514 DPSS available from 20 mW up to 150 mW available in the Sapphire form factor from Coherent. We use this laser, often in conjunction with the 445 CUBE in our LaserStack for CFP/YFP (and their newer brothers and sisters in FP evolution) imaging for FRET or otherwise. ------------------------ Samuel A. Connell Sales Manager Pacific Region-North America Intelligent Imaging Innovations, Inc 3250 Ocean Park Blvd, Suite 202 Santa Monica, CA 90405 Cell: (858) 692-4510 [hidden email] On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn .edu/cgi-bin/wa?A0=confocalmicroscopy> > ***** > > See: http://www.cobolt.se/**coboltfandango515nm.html?** > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw<http://www.cobolt.se/coboltfandango51 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> > > No connection, just considering about switching away from our old Ar Ion > as well. > > - Damir > > > On 11/9/2011 3:36 PM, Guy Cox wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> .edu/cgi-bin/wa?A0=confocalmicroscopy> >> ***** >> >> Are you sure there is a 514nm DPSS? I think that wavelength would have >> to be an OPS and I don't know anyone who makes one. Coherent do make a >> 505nm OPS which is intended to stand in for both the 488& 514 lines of >> >> an Argon laser, and I rather suspect that this could be exactly what you >> need. But I have no experience of it. (I do have a Coherent 488nm >> OPS). >> >> Guy >> >> Optical Imaging Techniques in Cell Biology >> by Guy Cox CRC Press / Taylor& Francis >> >> http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> >> ______________________________**________________ >> Associate Professor Guy Cox, MA, DPhil(Oxon) >> Australian Centre for Microscopy& Microanalysis, >> >> Madsen Building F09, University of Sydney, NSW 2006 >> >> Phone +61 2 9351 3176 Fax +61 2 9351 7682 >> Mobile 0413 281 861 >> ______________________________**________________ >> http://www.guycox.net >> >> >> >> -----Original Message----- >> From: Confocal Microscopy List EDU> >> ] >> On Behalf Of Tim Feinstein >> Sent: Thursday, 10 November 2011 7:34 AM >> To: [hidden email].**EDU <[hidden email]> >> Subject: FRET in the time of DPSS >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn .edu/cgi-bin/wa?A0=confocalmicroscopy> >> ***** >> >> Hello all, >> >> We want to spec a four-laser launch for a new live cell system that will >> handle both CFP/YFP FRET and red/green imaging. However, I am sad to >> see that gas lasers are no longer speccable and so the freebie 514 laser >> line is gone. We would therefore have to spec a 514 DPSS and forego the >> far-red line. >> >> I was wondering whether there is a way to do more (or at least the same) >> with less. 488 nm excites YFP well enough, so in theory I could image >> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm laser >> lines. In my experience 442 nm laser excitation (via TIRF) causes >> negligible YFP excitation and 488 nm does not excite CFP, so it is >> possible that I could gain speed by passing everything through a single >> broad bandpass filter (e.g., 455-550 nm) and alternate excitations. >> Assuming that cross-talk is not a problem, the most significant cost >> would be that I lose a decent chunk of CFP emission to the scan head >> dichroic, but in return I gain a 641 nm laser. >> >> Has anyone tried this? Any feedback on or off-list would be much >> appreciated. >> >> Thanks and all the best, >> >> >> TF >> >> Timothy Feinstein, PhD >> Postdoctoral Fellow >> Laboratory for GPCR Biology >> Dept. of Pharmacology& Chemical Biology >> >> University of Pittsburgh, School of Medicine >> BST W1301, 200 Lothrop St. >> Pittsburgh, PA 15261 >> >> ----- >> No virus found in this message. >> Checked by AVG - www.avg.com >> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: >> > > -- > Damir Sudar - Staff Scientist and Deputy for Technology > Lawrence Berkeley Laboratory / Life Sciences Division > One Cyclotron Road, MS 977, Berkeley, CA 94720, USA > T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] > WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** > Staff_Directory/Sudar_Lab.html<http://www.lbl.gov/lsd/People_&_Organizat ion/Scientific_Staff_Directory/Sudar_Lab.html> > ----- No virus found in this message. Checked by AVG - www.avg.com Version: 10.0.1411 / Virus Database: 2092/4006 - Release Date: 11/09/11 |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi guys (and Guy), Many thanks for the feedback. I like the 505 laser idea in particular. As long as it excites 488 fluorophores well enough it could be exactly what I need. Does anybody have experience with using that in place of a 488 line? All the best, Tim Sent from my iPad On Nov 10, 2011, at 1:57 AM, Guy Cox <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > The Sapphire series lasers are OPS (Optically Pumped Semiconductor) not > DPSS but, you are right, they do indeed have a 514nm version. I don't > know why I didn't find it, but did find the 505, when I searched this > morning. I still think that the 505 might be exactly what Timothy > needs, though, since he doesn't want to buy an extra laser. > > I have also been contacted off-list and told that Spectra-Physics have a > 515nm DPSS which is close enough to make no difference. I've no idea > what the lasing crystal is. > > Guy > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) > Australian Centre for Microscopy & Microanalysis, > Madsen Building F09, University of Sydney, NSW 2006 > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of samuel connell > Sent: Thursday, 10 November 2011 1:57 PM > To: [hidden email] > Subject: Re: FRET in the time of DPSS > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Industry Response: > > There absolutely is a 514 DPSS available from 20 mW up to 150 mW > available > in the Sapphire form factor from Coherent. We use this laser, often in > conjunction with the 445 CUBE in our LaserStack for CFP/YFP (and their > newer brothers and sisters in FP evolution) imaging for FRET or > otherwise. > > ------------------------ > Samuel A. Connell > Sales Manager > Pacific Region-North America > Intelligent Imaging Innovations, Inc > 3250 Ocean Park Blvd, Suite 202 > Santa Monica, CA 90405 > Cell: (858) 692-4510 > [hidden email] > > > On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn > .edu/cgi-bin/wa?A0=confocalmicroscopy> >> ***** >> >> See: http://www.cobolt.se/**coboltfandango515nm.html?** >> > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw<http://www.cobolt.se/coboltfandango51 > 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> >> >> No connection, just considering about switching away from our old Ar > Ion >> as well. >> >> - Damir >> >> >> On 11/9/2011 3:36 PM, Guy Cox wrote: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn > .edu/cgi-bin/wa?A0=confocalmicroscopy> >>> ***** >>> >>> Are you sure there is a 514nm DPSS? I think that wavelength would > have >>> to be an OPS and I don't know anyone who makes one. Coherent do make > a >>> 505nm OPS which is intended to stand in for both the 488& 514 lines > of >>> >>> an Argon laser, and I rather suspect that this could be exactly what > you >>> need. But I have no experience of it. (I do have a Coherent 488nm >>> OPS). >>> >>> Guy >>> >>> Optical Imaging Techniques in Cell Biology >>> by Guy Cox CRC Press / Taylor& Francis >>> >>> > http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> >>> ______________________________**________________ >>> Associate Professor Guy Cox, MA, DPhil(Oxon) >>> Australian Centre for Microscopy& Microanalysis, >>> >>> Madsen Building F09, University of Sydney, NSW 2006 >>> >>> Phone +61 2 9351 3176 Fax +61 2 9351 7682 >>> Mobile 0413 281 861 >>> ______________________________**________________ >>> http://www.guycox.net >>> >>> >>> >>> -----Original Message----- >>> From: Confocal Microscopy List > [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU<[hidden email]. > EDU> >>> ] >>> On Behalf Of Tim Feinstein >>> Sent: Thursday, 10 November 2011 7:34 AM >>> To: [hidden email].**EDU > <[hidden email]> >>> Subject: FRET in the time of DPSS >>> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn > .edu/cgi-bin/wa?A0=confocalmicroscopy> >>> ***** >>> >>> Hello all, >>> >>> We want to spec a four-laser launch for a new live cell system that > will >>> handle both CFP/YFP FRET and red/green imaging. However, I am sad > to >>> see that gas lasers are no longer speccable and so the freebie 514 > laser >>> line is gone. We would therefore have to spec a 514 DPSS and forego > the >>> far-red line. >>> >>> I was wondering whether there is a way to do more (or at least the > same) >>> with less. 488 nm excites YFP well enough, so in theory I could > image >>> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm > laser >>> lines. In my experience 442 nm laser excitation (via TIRF) causes >>> negligible YFP excitation and 488 nm does not excite CFP, so it is >>> possible that I could gain speed by passing everything through a > single >>> broad bandpass filter (e.g., 455-550 nm) and alternate excitations. >>> Assuming that cross-talk is not a problem, the most significant cost >>> would be that I lose a decent chunk of CFP emission to the scan head >>> dichroic, but in return I gain a 641 nm laser. >>> >>> Has anyone tried this? Any feedback on or off-list would be much >>> appreciated. >>> >>> Thanks and all the best, >>> >>> >>> TF >>> >>> Timothy Feinstein, PhD >>> Postdoctoral Fellow >>> Laboratory for GPCR Biology >>> Dept. of Pharmacology& Chemical Biology >>> >>> University of Pittsburgh, School of Medicine >>> BST W1301, 200 Lothrop St. >>> Pittsburgh, PA 15261 >>> >>> ----- >>> No virus found in this message. >>> Checked by AVG - www.avg.com >>> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: > 11/08/11 >>> >> >> -- >> Damir Sudar - Staff Scientist and Deputy for Technology >> Lawrence Berkeley Laboratory / Life Sciences Division >> One Cyclotron Road, MS 977, Berkeley, CA 94720, USA >> T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] >> WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** >> > Staff_Directory/Sudar_Lab.html<http://www.lbl.gov/lsd/People_&_Organizat > ion/Scientific_Staff_Directory/Sudar_Lab.html> >> > > ----- > No virus found in this message. > Checked by AVG - www.avg.com > Version: 10.0.1411 / Virus Database: 2092/4006 - Release Date: 11/09/11 |
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Boswell, Carl A - (cboswell) |
Re: FRET in the time of DPSS
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Hi Tim,
The spectral website: http://www.spectra.arizona.edu/ will show you specifically what the "efficiency" of absorbance is for any dye you might be considering. Simply place the cursor over the curve at any wavelength to find out the relative absorbance. This can be used to roughly pre-test dyes vs. excitation sources so you have a rational basis for considering which laser might do what you need. Good luck. C Carl A. Boswell 520-954-7053 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Tim Feinstein Sent: Thursday, November 10, 2011 6:09 AM To: [hidden email] Subject: Re: FRET in the time of DPSS ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi guys (and Guy), Many thanks for the feedback. I like the 505 laser idea in particular. As long as it excites 488 fluorophores well enough it could be exactly what I need. Does anybody have experience with using that in place of a 488 line? All the best, Tim Sent from my iPad On Nov 10, 2011, at 1:57 AM, Guy Cox <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > The Sapphire series lasers are OPS (Optically Pumped Semiconductor) > not DPSS but, you are right, they do indeed have a 514nm version. I > don't know why I didn't find it, but did find the 505, when I searched > this morning. I still think that the 505 might be exactly what > Timothy needs, though, since he doesn't want to buy an extra laser. > > I have also been contacted off-list and told that Spectra-Physics have > a 515nm DPSS which is close enough to make no difference. I've no > idea what the lasing crystal is. > > Guy > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) Australian Centre for > Microscopy & Microanalysis, Madsen Building F09, University of Sydney, > NSW 2006 > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > > > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] > On Behalf Of samuel connell > Sent: Thursday, 10 November 2011 1:57 PM > To: [hidden email] > Subject: Re: FRET in the time of DPSS > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Industry Response: > > There absolutely is a 514 DPSS available from 20 mW up to 150 mW > available in the Sapphire form factor from Coherent. We use this > laser, often in conjunction with the 445 CUBE in our LaserStack for > CFP/YFP (and their newer brothers and sisters in FP evolution) imaging > for FRET or otherwise. > > ------------------------ > Samuel A. Connell > Sales Manager > Pacific Region-North America > Intelligent Imaging Innovations, Inc > 3250 Ocean Park Blvd, Suite 202 > Santa Monica, CA 90405 > Cell: (858) 692-4510 > [hidden email] > > > On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.u > mn .edu/cgi-bin/wa?A0=confocalmicroscopy> >> ***** >> >> See: http://www.cobolt.se/**coboltfandango515nm.html?** >> > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw<http://www.cobolt.se/coboltfandango > 51 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> >> >> No connection, just considering about switching away from our old Ar > Ion >> as well. >> >> - Damir >> >> >> On 11/9/2011 3:36 PM, Guy Cox wrote: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.u > mn .edu/cgi-bin/wa?A0=confocalmicroscopy> >>> ***** >>> >>> Are you sure there is a 514nm DPSS? I think that wavelength would > have >>> to be an OPS and I don't know anyone who makes one. Coherent do >>> make > a >>> 505nm OPS which is intended to stand in for both the 488& 514 lines > of >>> >>> an Argon laser, and I rather suspect that this could be exactly what > you >>> need. But I have no experience of it. (I do have a Coherent 488nm >>> OPS). >>> >>> Guy >>> >>> Optical Imaging Techniques in Cell Biology >>> by Guy Cox CRC Press / Taylor& Francis >>> >>> > http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> >>> ______________________________**________________ >>> Associate Professor Guy Cox, MA, DPhil(Oxon) Australian Centre for >>> Microscopy& Microanalysis, >>> >>> Madsen Building F09, University of Sydney, NSW 2006 >>> >>> Phone +61 2 9351 3176 Fax +61 2 9351 7682 >>> Mobile 0413 281 861 >>> ______________________________**________________ >>> http://www.guycox.net >>> >>> >>> >>> -----Original Message----- >>> From: Confocal Microscopy List > [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU<[hidden email]. > EDU> >>> ] >>> On Behalf Of Tim Feinstein >>> Sent: Thursday, 10 November 2011 7:34 AM >>> To: [hidden email].**EDU > <[hidden email]> >>> Subject: FRET in the time of DPSS >>> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.u > mn .edu/cgi-bin/wa?A0=confocalmicroscopy> >>> ***** >>> >>> Hello all, >>> >>> We want to spec a four-laser launch for a new live cell system that > will >>> handle both CFP/YFP FRET and red/green imaging. However, I am sad > to >>> see that gas lasers are no longer speccable and so the freebie 514 > laser >>> line is gone. We would therefore have to spec a 514 DPSS and forego > the >>> far-red line. >>> >>> I was wondering whether there is a way to do more (or at least the > same) >>> with less. 488 nm excites YFP well enough, so in theory I could > image >>> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm > laser >>> lines. In my experience 442 nm laser excitation (via TIRF) causes >>> negligible YFP excitation and 488 nm does not excite CFP, so it is >>> possible that I could gain speed by passing everything through a > single >>> broad bandpass filter (e.g., 455-550 nm) and alternate excitations. >>> Assuming that cross-talk is not a problem, the most significant cost >>> would be that I lose a decent chunk of CFP emission to the scan head >>> dichroic, but in return I gain a 641 nm laser. >>> >>> Has anyone tried this? Any feedback on or off-list would be much >>> appreciated. >>> >>> Thanks and all the best, >>> >>> >>> TF >>> >>> Timothy Feinstein, PhD >>> Postdoctoral Fellow >>> Laboratory for GPCR Biology >>> Dept. of Pharmacology& Chemical Biology >>> >>> University of Pittsburgh, School of Medicine BST W1301, 200 Lothrop >>> St. >>> Pittsburgh, PA 15261 >>> >>> ----- >>> No virus found in this message. >>> Checked by AVG - www.avg.com >>> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: > 11/08/11 >>> >> >> -- >> Damir Sudar - Staff Scientist and Deputy for Technology Lawrence >> Berkeley Laboratory / Life Sciences Division One Cyclotron Road, MS >> 977, Berkeley, CA 94720, USA >> T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] >> WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** >> > Staff_Directory/Sudar_Lab.html<http://www.lbl.gov/lsd/People_&_Organiz > at ion/Scientific_Staff_Directory/Sudar_Lab.html> >> > > ----- > No virus found in this message. > Checked by AVG - www.avg.com > Version: 10.0.1411 / Virus Database: 2092/4006 - Release Date: > 11/09/11 |
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In reply to this post by Tim Feinstein-2
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** There are also the new direct diode lasers at 520nm if you can handle a 6nm red wavelength shift from 514. They've only been around for about a year but many manufacturers have lumped them in with their direct-diode product lines and they look pretty decent. Craig On Thu, Nov 10, 2011 at 6:08 AM, Tim Feinstein <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi guys (and Guy), > > Many thanks for the feedback. I like the 505 laser idea in particular. > As long as it excites 488 fluorophores well enough it could be exactly > what I need. Does anybody have experience with using that in place of a > 488 line? > > All the best, > > > Tim > > Sent from my iPad > > On Nov 10, 2011, at 1:57 AM, Guy Cox <[hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > The Sapphire series lasers are OPS (Optically Pumped Semiconductor) not > > DPSS but, you are right, they do indeed have a 514nm version. I don't > > know why I didn't find it, but did find the 505, when I searched this > > morning. I still think that the 505 might be exactly what Timothy > > needs, though, since he doesn't want to buy an extra laser. > > > > I have also been contacted off-list and told that Spectra-Physics have a > > 515nm DPSS which is close enough to make no difference. I've no idea > > what the lasing crystal is. > > > > Guy > > > > Optical Imaging Techniques in Cell Biology > > by Guy Cox CRC Press / Taylor & Francis > > http://www.guycox.com/optical.htm > > ______________________________________________ > > Associate Professor Guy Cox, MA, DPhil(Oxon) > > Australian Centre for Microscopy & Microanalysis, > > Madsen Building F09, University of Sydney, NSW 2006 > > > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > Mobile 0413 281 861 > > ______________________________________________ > > http://www.guycox.net > > > > > > > > -----Original Message----- > > From: Confocal Microscopy List [mailto:[hidden email]] > > On Behalf Of samuel connell > > Sent: Thursday, 10 November 2011 1:57 PM > > To: [hidden email] > > Subject: Re: FRET in the time of DPSS > > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Industry Response: > > > > There absolutely is a 514 DPSS available from 20 mW up to 150 mW > > available > > in the Sapphire form factor from Coherent. We use this laser, often in > > conjunction with the 445 CUBE in our LaserStack for CFP/YFP (and their > > newer brothers and sisters in FP evolution) imaging for FRET or > > otherwise. > > > > ------------------------ > > Samuel A. Connell > > Sales Manager > > Pacific Region-North America > > Intelligent Imaging Innovations, Inc > > 3250 Ocean Park Blvd, Suite 202 > > Santa Monica, CA 90405 > > Cell: (858) 692-4510 > > [hidden email] > > > > > > On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> wrote: > > > >> ***** > >> To join, leave or search the confocal microscopy listserv, go to: > >> > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > >> ***** > >> > >> See: http://www.cobolt.se/**coboltfandango515nm.html?** > >> > > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw<http://www.cobolt.se/coboltfandango51 > > 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> > >> > >> No connection, just considering about switching away from our old Ar > > Ion > >> as well. > >> > >> - Damir > >> > >> > >> On 11/9/2011 3:36 PM, Guy Cox wrote: > >> > >>> ***** > >>> To join, leave or search the confocal microscopy listserv, go to: > >>> > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > >>> ***** > >>> > >>> Are you sure there is a 514nm DPSS? I think that wavelength would > > have > >>> to be an OPS and I don't know anyone who makes one. Coherent do make > > a > >>> 505nm OPS which is intended to stand in for both the 488& 514 lines > > of > >>> > >>> an Argon laser, and I rather suspect that this could be exactly what > > you > >>> need. But I have no experience of it. (I do have a Coherent 488nm > >>> OPS). > >>> > >>> Guy > >>> > >>> Optical Imaging Techniques in Cell Biology > >>> by Guy Cox CRC Press / Taylor& Francis > >>> > >>> > > http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> > >>> ______________________________**________________ > >>> Associate Professor Guy Cox, MA, DPhil(Oxon) > >>> Australian Centre for Microscopy& Microanalysis, > >>> > >>> Madsen Building F09, University of Sydney, NSW 2006 > >>> > >>> Phone +61 2 9351 3176 Fax +61 2 9351 7682 > >>> Mobile 0413 281 861 > >>> ______________________________**________________ > >>> http://www.guycox.net > >>> > >>> > >>> > >>> -----Original Message----- > >>> From: Confocal Microscopy List > > [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU<[hidden email]. > > EDU> > >>> ] > >>> On Behalf Of Tim Feinstein > >>> Sent: Thursday, 10 November 2011 7:34 AM > >>> To: [hidden email].**EDU > > <[hidden email]> > >>> Subject: FRET in the time of DPSS > >>> > >>> ***** > >>> To join, leave or search the confocal microscopy listserv, go to: > >>> > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > >>> ***** > >>> > >>> Hello all, > >>> > >>> We want to spec a four-laser launch for a new live cell system that > > will > >>> handle both CFP/YFP FRET and red/green imaging. However, I am sad > > to > >>> see that gas lasers are no longer speccable and so the freebie 514 > > laser > >>> line is gone. We would therefore have to spec a 514 DPSS and forego > > the > >>> far-red line. > >>> > >>> I was wondering whether there is a way to do more (or at least the > > same) > >>> with less. 488 nm excites YFP well enough, so in theory I could > > image > >>> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm > > laser > >>> lines. In my experience 442 nm laser excitation (via TIRF) causes > >>> negligible YFP excitation and 488 nm does not excite CFP, so it is > >>> possible that I could gain speed by passing everything through a > > single > >>> broad bandpass filter (e.g., 455-550 nm) and alternate excitations. > >>> Assuming that cross-talk is not a problem, the most significant cost > >>> would be that I lose a decent chunk of CFP emission to the scan head > >>> dichroic, but in return I gain a 641 nm laser. > >>> > >>> Has anyone tried this? Any feedback on or off-list would be much > >>> appreciated. > >>> > >>> Thanks and all the best, > >>> > >>> > >>> TF > >>> > >>> Timothy Feinstein, PhD > >>> Postdoctoral Fellow > >>> Laboratory for GPCR Biology > >>> Dept. of Pharmacology& Chemical Biology > >>> > >>> University of Pittsburgh, School of Medicine > >>> BST W1301, 200 Lothrop St. > >>> Pittsburgh, PA 15261 > >>> > >>> ----- > >>> No virus found in this message. > >>> Checked by AVG - www.avg.com > >>> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: > > 11/08/11 > >>> > >> > >> -- > >> Damir Sudar - Staff Scientist and Deputy for Technology > >> Lawrence Berkeley Laboratory / Life Sciences Division > >> One Cyclotron Road, MS 977, Berkeley, CA 94720, USA > >> T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] > >> WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** > >> > > Staff_Directory/Sudar_Lab.html<http://www.lbl.gov/lsd/People_&_Organizat > > ion/Scientific_Staff_Directory/Sudar_Lab.html> > >> > > > > ----- > > No virus found in this message. > > Checked by AVG - www.avg.com > > Version: 10.0.1411 / Virus Database: 2092/4006 - Release Date: 11/09/11 > |
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Hi all
This discussion is very timely for our facility, as we are looking to purchase a solide state laser in the ~500-520nm range as soon as possible I am insufficiently familiar with the pros and cons of choosing a DPSS versus a OPS or a direct-diode laser From the discussion so far it sounds like the choices for a solid state laser for imaging YFP are 515nm DPSS from Spectra-Physics, a 505nm Sapphire OPS, a 514nm Sapphire OPS or a 520 direct diode. Guy - I presume by "direct diode" you mean a diode pumped solid state, which I understand would be not as good as a DPSS (please correct me if I am wrong) I dont have any idea how an OPS compares to a DPSS laser Any advice as to which of these options might be "best" as an add on to a new Olympus FV1000 for the primary purpose of imaging and bleaching YFP in the CFP/YFP FRET pair would be greatly appreciated. Kind regards Jen -- Jennifer Clarke BSc (Hons) PhD Research Associate, Anatomy and Histology Centre for Neuroscience, School of Medicine & Facility Manager, Optical Microscopy Suite, Flinders Microscopy Flinders University GPO Box 2100, Adelaide 5001 Phone: 61 8 8204 6454/ 61 8 8204 6637 Email: [hidden email] ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Craig Brideau [[hidden email]] Sent: Friday, 11 November 2011 5:03 AM To: [hidden email] Subject: Re: FRET in the time of DPSS ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** There are also the new direct diode lasers at 520nm if you can handle a 6nm red wavelength shift from 514. They've only been around for about a year but many manufacturers have lumped them in with their direct-diode product lines and they look pretty decent. Craig On Thu, Nov 10, 2011 at 6:08 AM, Tim Feinstein <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi guys (and Guy), > > Many thanks for the feedback. I like the 505 laser idea in particular. > As long as it excites 488 fluorophores well enough it could be exactly > what I need. Does anybody have experience with using that in place of a > 488 line? > > All the best, > > > Tim > > Sent from my iPad > > On Nov 10, 2011, at 1:57 AM, Guy Cox <[hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > The Sapphire series lasers are OPS (Optically Pumped Semiconductor) not > > DPSS but, you are right, they do indeed have a 514nm version. I don't > > know why I didn't find it, but did find the 505, when I searched this > > morning. I still think that the 505 might be exactly what Timothy > > needs, though, since he doesn't want to buy an extra laser. > > > > I have also been contacted off-list and told that Spectra-Physics have a > > 515nm DPSS which is close enough to make no difference. I've no idea > > what the lasing crystal is. > > > > Guy > > > > Optical Imaging Techniques in Cell Biology > > by Guy Cox CRC Press / Taylor & Francis > > http://www.guycox.com/optical.htm > > ______________________________________________ > > Associate Professor Guy Cox, MA, DPhil(Oxon) > > Australian Centre for Microscopy & Microanalysis, > > Madsen Building F09, University of Sydney, NSW 2006 > > > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > Mobile 0413 281 861 > > ______________________________________________ > > http://www.guycox.net > > > > > > > > -----Original Message----- > > From: Confocal Microscopy List [mailto:[hidden email]] > > On Behalf Of samuel connell > > Sent: Thursday, 10 November 2011 1:57 PM > > To: [hidden email] > > Subject: Re: FRET in the time of DPSS > > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Industry Response: > > > > There absolutely is a 514 DPSS available from 20 mW up to 150 mW > > available > > in the Sapphire form factor from Coherent. We use this laser, often in > > conjunction with the 445 CUBE in our LaserStack for CFP/YFP (and their > > newer brothers and sisters in FP evolution) imaging for FRET or > > otherwise. > > > > ------------------------ > > Samuel A. Connell > > Sales Manager > > Pacific Region-North America > > Intelligent Imaging Innovations, Inc > > 3250 Ocean Park Blvd, Suite 202 > > Santa Monica, CA 90405 > > Cell: (858) 692-4510 > > [hidden email] > > > > > > On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> wrote: > > > >> ***** > >> To join, leave or search the confocal microscopy listserv, go to: > >> > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > >> ***** > >> > >> See: http://www.cobolt.se/**coboltfandango515nm.html?** > >> > > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw<http://www.cobolt.se/coboltfandango51 > > 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> > >> > >> No connection, just considering about switching away from our old Ar > > Ion > >> as well. > >> > >> - Damir > >> > >> > >> On 11/9/2011 3:36 PM, Guy Cox wrote: > >> > >>> ***** > >>> To join, leave or search the confocal microscopy listserv, go to: > >>> > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > >>> ***** > >>> > >>> Are you sure there is a 514nm DPSS? I think that wavelength would > > have > >>> to be an OPS and I don't know anyone who makes one. Coherent do make > > a > >>> 505nm OPS which is intended to stand in for both the 488& 514 lines > > of > >>> > >>> an Argon laser, and I rather suspect that this could be exactly what > > you > >>> need. But I have no experience of it. (I do have a Coherent 488nm > >>> OPS). > >>> > >>> Guy > >>> > >>> Optical Imaging Techniques in Cell Biology > >>> by Guy Cox CRC Press / Taylor& Francis > >>> > >>> > > http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> > >>> ______________________________**________________ > >>> Associate Professor Guy Cox, MA, DPhil(Oxon) > >>> Australian Centre for Microscopy& Microanalysis, > >>> > >>> Madsen Building F09, University of Sydney, NSW 2006 > >>> > >>> Phone +61 2 9351 3176 Fax +61 2 9351 7682 > >>> Mobile 0413 281 861 > >>> ______________________________**________________ > >>> http://www.guycox.net > >>> > >>> > >>> > >>> -----Original Message----- > >>> From: Confocal Microscopy List > > [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU<[hidden email]. > > EDU> > >>> ] > >>> On Behalf Of Tim Feinstein > >>> Sent: Thursday, 10 November 2011 7:34 AM > >>> To: [hidden email].**EDU > > <[hidden email]> > >>> Subject: FRET in the time of DPSS > >>> > >>> ***** > >>> To join, leave or search the confocal microscopy listserv, go to: > >>> > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.umn > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > >>> ***** > >>> > >>> Hello all, > >>> > >>> We want to spec a four-laser launch for a new live cell system that > > will > >>> handle both CFP/YFP FRET and red/green imaging. However, I am sad > > to > >>> see that gas lasers are no longer speccable and so the freebie 514 > > laser > >>> line is gone. We would therefore have to spec a 514 DPSS and forego > > the > >>> far-red line. > >>> > >>> I was wondering whether there is a way to do more (or at least the > > same) > >>> with less. 488 nm excites YFP well enough, so in theory I could > > image > >>> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm > > laser > >>> lines. In my experience 442 nm laser excitation (via TIRF) causes > >>> negligible YFP excitation and 488 nm does not excite CFP, so it is > >>> possible that I could gain speed by passing everything through a > > single > >>> broad bandpass filter (e.g., 455-550 nm) and alternate excitations. > >>> Assuming that cross-talk is not a problem, the most significant cost > >>> would be that I lose a decent chunk of CFP emission to the scan head > >>> dichroic, but in return I gain a 641 nm laser. > >>> > >>> Has anyone tried this? Any feedback on or off-list would be much > >>> appreciated. > >>> > >>> Thanks and all the best, > >>> > >>> > >>> TF > >>> > >>> Timothy Feinstein, PhD > >>> Postdoctoral Fellow > >>> Laboratory for GPCR Biology > >>> Dept. of Pharmacology& Chemical Biology > >>> > >>> University of Pittsburgh, School of Medicine > >>> BST W1301, 200 Lothrop St. > >>> Pittsburgh, PA 15261 > >>> > >>> ----- > >>> No virus found in this message. > >>> Checked by AVG - www.avg.com > >>> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: > > 11/08/11 > >>> > >> > >> -- > >> Damir Sudar - Staff Scientist and Deputy for Technology > >> Lawrence Berkeley Laboratory / Life Sciences Division > >> One Cyclotron Road, MS 977, Berkeley, CA 94720, USA > >> T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] > >> WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** > >> > > Staff_Directory/Sudar_Lab.html<http://www.lbl.gov/lsd/People_&_Organizat > > ion/Scientific_Staff_Directory/Sudar_Lab.html> > >> > > > > ----- > > No virus found in this message. > > Checked by AVG - www.avg.com > > Version: 10.0.1411 / Virus Database: 2092/4006 - Release Date: 11/09/11 > |
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*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Direct diode means that the laser has a single element. It tends to be the simplest type of laser; basically it is a big laser pointer. Diode-pumped solid state use an infrared diode (typically) which pumps some secondary medium that gives you your desired wavelength. They are more complex than direct diode since you now have two separate objects that have to be reliably optically coupled. Until recently direct diode lasers have only been capable of near-IR, Red, and Violet or Blue lines, but a recent breakthrough in laser diode chemistry has allowed them to work at around 520nm. Here's an old article from 2009 mentioning one of the first green direct diode lasers: http://displaydaily.com/2009/07/21/green-laser-diode-demonstrated-a-breakthrough/ The current problem is pushing them to longer wavelengths. Projector companies want to get them out to 530, 540nm for making displays and video projectors: http://www.qmed.com/mpmn/article/suppliers-chip-based-diode-makes-waves-green-laser-world But 'shorter' direct diode green lasers in the 520nm or less range are fairly available right now. Craig On Thu, Nov 10, 2011 at 5:09 PM, Jen Clarke <[hidden email] > wrote: > Hi all > > This discussion is very timely for our facility, as we are looking to > purchase a solide state laser in the ~500-520nm range as soon as possible > > I am insufficiently familiar with the pros and cons of choosing a DPSS > versus a OPS or a direct-diode laser > > From the discussion so far it sounds like the choices for a solid state > laser for imaging YFP are 515nm DPSS from Spectra-Physics, a 505nm Sapphire > OPS, a 514nm Sapphire OPS or a 520 direct diode. > > Guy - I presume by "direct diode" you mean a diode pumped solid state, > which I understand would be not as good as a DPSS (please correct me if I > am wrong) > > I dont have any idea how an OPS compares to a DPSS laser > > Any advice as to which of these options might be "best" as an add on to a > new Olympus FV1000 for the primary purpose of imaging and bleaching YFP in > the CFP/YFP FRET pair would be greatly appreciated. > > Kind regards > Jen > -- > Jennifer Clarke BSc (Hons) PhD > Research Associate, Anatomy and Histology > Centre for Neuroscience, School of Medicine > & > Facility Manager, Optical Microscopy Suite, Flinders Microscopy > > Flinders University > GPO Box 2100, Adelaide 5001 > Phone: 61 8 8204 6454/ 61 8 8204 6637 > Email: [hidden email] > > > ________________________________________ > From: Confocal Microscopy List [[hidden email]] On > Behalf Of Craig Brideau [[hidden email]] > Sent: Friday, 11 November 2011 5:03 AM > To: [hidden email] > Subject: Re: FRET in the time of DPSS > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > There are also the new direct diode lasers at 520nm if you can handle a 6nm > red wavelength shift from 514. They've only been around for about a year > but many manufacturers have lumped them in with their direct-diode product > lines and they look pretty decent. > > Craig > > > > On Thu, Nov 10, 2011 at 6:08 AM, Tim Feinstein <[hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Hi guys (and Guy), > > > > Many thanks for the feedback. I like the 505 laser idea in particular. > > As long as it excites 488 fluorophores well enough it could be exactly > > what I need. Does anybody have experience with using that in place of a > > 488 line? > > > > All the best, > > > > > > Tim > > > > Sent from my iPad > > > > On Nov 10, 2011, at 1:57 AM, Guy Cox <[hidden email]> wrote: > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > The Sapphire series lasers are OPS (Optically Pumped Semiconductor) not > > > DPSS but, you are right, they do indeed have a 514nm version. I don't > > > know why I didn't find it, but did find the 505, when I searched this > > > morning. I still think that the 505 might be exactly what Timothy > > > needs, though, since he doesn't want to buy an extra laser. > > > > > > I have also been contacted off-list and told that Spectra-Physics have > a > > > 515nm DPSS which is close enough to make no difference. I've no idea > > > what the lasing crystal is. > > > > > > Guy > > > > > > Optical Imaging Techniques in Cell Biology > > > by Guy Cox CRC Press / Taylor & Francis > > > http://www.guycox.com/optical.htm > > > ______________________________________________ > > > Associate Professor Guy Cox, MA, DPhil(Oxon) > > > Australian Centre for Microscopy & Microanalysis, > > > Madsen Building F09, University of Sydney, NSW 2006 > > > > > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > > Mobile 0413 281 861 > > > ______________________________________________ > > > http://www.guycox.net > > > > > > > > > > > > -----Original Message----- > > > From: Confocal Microscopy List [mailto: > [hidden email]] > > > On Behalf Of samuel connell > > > Sent: Thursday, 10 November 2011 1:57 PM > > > To: [hidden email] > > > Subject: Re: FRET in the time of DPSS > > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > Industry Response: > > > > > > There absolutely is a 514 DPSS available from 20 mW up to 150 mW > > > available > > > in the Sapphire form factor from Coherent. We use this laser, often in > > > conjunction with the 445 CUBE in our LaserStack for CFP/YFP (and their > > > newer brothers and sisters in FP evolution) imaging for FRET or > > > otherwise. > > > > > > ------------------------ > > > Samuel A. Connell > > > Sales Manager > > > Pacific Region-North America > > > Intelligent Imaging Innovations, Inc > > > 3250 Ocean Park Blvd, Suite 202 > > > Santa Monica, CA 90405 > > > Cell: (858) 692-4510 > > > [hidden email] > > > > > > > > > On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> wrote: > > > > > >> ***** > > >> To join, leave or search the confocal microscopy listserv, go to: > > >> > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > http://lists.umn > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > >> ***** > > >> > > >> See: http://www.cobolt.se/**coboltfandango515nm.html?** > > >> > > > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw< > http://www.cobolt.se/coboltfandango51 > > > 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> > > >> > > >> No connection, just considering about switching away from our old Ar > > > Ion > > >> as well. > > >> > > >> - Damir > > >> > > >> > > >> On 11/9/2011 3:36 PM, Guy Cox wrote: > > >> > > >>> ***** > > >>> To join, leave or search the confocal microscopy listserv, go to: > > >>> > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > http://lists.umn > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > >>> ***** > > >>> > > >>> Are you sure there is a 514nm DPSS? I think that wavelength would > > > have > > >>> to be an OPS and I don't know anyone who makes one. Coherent do make > > > a > > >>> 505nm OPS which is intended to stand in for both the 488& 514 lines > > > of > > >>> > > >>> an Argon laser, and I rather suspect that this could be exactly what > > > you > > >>> need. But I have no experience of it. (I do have a Coherent 488nm > > >>> OPS). > > >>> > > >>> Guy > > >>> > > >>> Optical Imaging Techniques in Cell Biology > > >>> by Guy Cox CRC Press / Taylor& Francis > > >>> > > >>> > > > http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> > > >>> ______________________________**________________ > > >>> Associate Professor Guy Cox, MA, DPhil(Oxon) > > >>> Australian Centre for Microscopy& Microanalysis, > > >>> > > >>> Madsen Building F09, University of Sydney, NSW 2006 > > >>> > > >>> Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > >>> Mobile 0413 281 861 > > >>> ______________________________**________________ > > >>> http://www.guycox.net > > >>> > > >>> > > >>> > > >>> -----Original Message----- > > >>> From: Confocal Microscopy List > > > [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU > <[hidden email]. > > > EDU> > > >>> ] > > >>> On Behalf Of Tim Feinstein > > >>> Sent: Thursday, 10 November 2011 7:34 AM > > >>> To: [hidden email].**EDU > > > <[hidden email]> > > >>> Subject: FRET in the time of DPSS > > >>> > > >>> ***** > > >>> To join, leave or search the confocal microscopy listserv, go to: > > >>> > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > http://lists.umn > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > >>> ***** > > >>> > > >>> Hello all, > > >>> > > >>> We want to spec a four-laser launch for a new live cell system that > > > will > > >>> handle both CFP/YFP FRET and red/green imaging. However, I am sad > > > to > > >>> see that gas lasers are no longer speccable and so the freebie 514 > > > laser > > >>> line is gone. We would therefore have to spec a 514 DPSS and forego > > > the > > >>> far-red line. > > >>> > > >>> I was wondering whether there is a way to do more (or at least the > > > same) > > >>> with less. 488 nm excites YFP well enough, so in theory I could > > > image > > >>> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm > > > laser > > >>> lines. In my experience 442 nm laser excitation (via TIRF) causes > > >>> negligible YFP excitation and 488 nm does not excite CFP, so it is > > >>> possible that I could gain speed by passing everything through a > > > single > > >>> broad bandpass filter (e.g., 455-550 nm) and alternate excitations. > > >>> Assuming that cross-talk is not a problem, the most significant cost > > >>> would be that I lose a decent chunk of CFP emission to the scan head > > >>> dichroic, but in return I gain a 641 nm laser. > > >>> > > >>> Has anyone tried this? Any feedback on or off-list would be much > > >>> appreciated. > > >>> > > >>> Thanks and all the best, > > >>> > > >>> > > >>> TF > > >>> > > >>> Timothy Feinstein, PhD > > >>> Postdoctoral Fellow > > >>> Laboratory for GPCR Biology > > >>> Dept. of Pharmacology& Chemical Biology > > >>> > > >>> University of Pittsburgh, School of Medicine > > >>> BST W1301, 200 Lothrop St. > > >>> Pittsburgh, PA 15261 > > >>> > > >>> ----- > > >>> No virus found in this message. > > >>> Checked by AVG - www.avg.com > > >>> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: > > > 11/08/11 > > >>> > > >> > > >> -- > > >> Damir Sudar - Staff Scientist and Deputy for Technology > > >> Lawrence Berkeley Laboratory / Life Sciences Division > > >> One Cyclotron Road, MS 977, Berkeley, CA 94720, USA > > >> T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] > > >> WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** > > >> > > > Staff_Directory/Sudar_Lab.html< > http://www.lbl.gov/lsd/People_&_Organizat > > > ion/Scientific_Staff_Directory/Sudar_Lab.html> > > >> > > > > > > ----- > > > No virus found in this message. > > > Checked by AVG - www.avg.com > > > Version: 10.0.1411 / Virus Database: 2092/4006 - Release Date: 11/09/11 > > > > |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** OK, this is quite a confusing area (and the manufacturers do not always make things as clear as they could). A simple diode laser is a pure semiconductor device. The laser cavity is contained within a single chip (basically an LED with mirror layers above and below it). Wavelengths currently available are from near UV (~365nm) through violet to blue (~470nm), and then down in the red from ~640nm. Because the laser cavity is so small the wavelength is not very precise, which can be a problem for AOD based devices, though it is not likely to be a biological problem. Your 405 and 440 nm lasers are most likely simple diodes, as well and your 647nm. If you have a red laser pointer this will be a simple diode. OPS (optically pumped semiconductor) lasers are semiconductor devices which are not excited by electric current but by input from a diode laser. They can access wavelengths which direct diodes cannot. I think it is fair to say that this has been very much Coherent technology. 488 - 515 lasers, which are tricky from other sources, are within this range. DPSS are solid state crystal lasers pumped by red diode lasers. Neodymium Yttrium Aluminium Garnet (NdYAG) is the classic, working at 1064nm. For our purposes we are mostly interested in lasers which are frequency doubled by second harmonic generating crystals and the classic is therefore 532nm - the wavelength of your green laser pointer and the laser which pumps your TiS laser for multiphoton microscopy. Ytterbium lasers give 1030 nm (and hence 515nm when doubled). 561 nm is another option, and I believe it comes from a secondary line from an Nd YAG. Correction from laser companies welcome, but I hope this helps. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Australian Centre for Microscopy & Microanalysis, Madsen Building F09, University of Sydney, NSW 2006 Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Craig Brideau Sent: Friday, 11 November 2011 12:05 PM To: [hidden email] Subject: Re: FRET in the time of DPSS ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Direct diode means that the laser has a single element. It tends to be the simplest type of laser; basically it is a big laser pointer. Diode-pumped solid state use an infrared diode (typically) which pumps some secondary medium that gives you your desired wavelength. They are more complex than direct diode since you now have two separate objects that have to be reliably optically coupled. Until recently direct diode lasers have only been capable of near-IR, Red, and Violet or Blue lines, but a recent breakthrough in laser diode chemistry has allowed them to work at around 520nm. Here's an old article from 2009 mentioning one of the first green direct diode lasers: http://displaydaily.com/2009/07/21/green-laser-diode-demonstrated-a-brea kthrough/ The current problem is pushing them to longer wavelengths. Projector companies want to get them out to 530, 540nm for making displays and video projectors: http://www.qmed.com/mpmn/article/suppliers-chip-based-diode-makes-waves- green-laser-world But 'shorter' direct diode green lasers in the 520nm or less range are fairly available right now. Craig On Thu, Nov 10, 2011 at 5:09 PM, Jen Clarke <[hidden email] > wrote: > Hi all > > This discussion is very timely for our facility, as we are looking to > purchase a solide state laser in the ~500-520nm range as soon as possible > > I am insufficiently familiar with the pros and cons of choosing a DPSS > versus a OPS or a direct-diode laser > > From the discussion so far it sounds like the choices for a solid state > laser for imaging YFP are 515nm DPSS from Spectra-Physics, a 505nm Sapphire > OPS, a 514nm Sapphire OPS or a 520 direct diode. > > Guy - I presume by "direct diode" you mean a diode pumped solid state, > which I understand would be not as good as a DPSS (please correct me if I > am wrong) > > I dont have any idea how an OPS compares to a DPSS laser > > Any advice as to which of these options might be "best" as an add on to a > new Olympus FV1000 for the primary purpose of imaging and bleaching YFP in > the CFP/YFP FRET pair would be greatly appreciated. > > Kind regards > Jen > -- > Jennifer Clarke BSc (Hons) PhD > Research Associate, Anatomy and Histology > Centre for Neuroscience, School of Medicine > & > Facility Manager, Optical Microscopy Suite, Flinders Microscopy > > Flinders University > GPO Box 2100, Adelaide 5001 > Phone: 61 8 8204 6454/ 61 8 8204 6637 > Email: [hidden email] > > > ________________________________________ > From: Confocal Microscopy List [[hidden email]] On > Behalf Of Craig Brideau [[hidden email]] > Sent: Friday, 11 November 2011 5:03 AM > To: [hidden email] > Subject: Re: FRET in the time of DPSS > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > There are also the new direct diode lasers at 520nm if you can handle > red wavelength shift from 514. They've only been around for about a year > but many manufacturers have lumped them in with their direct-diode product > lines and they look pretty decent. > > Craig > > > > On Thu, Nov 10, 2011 at 6:08 AM, Tim Feinstein <[hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Hi guys (and Guy), > > > > Many thanks for the feedback. I like the 505 laser idea in > > As long as it excites 488 fluorophores well enough it could be exactly > > what I need. Does anybody have experience with using that in place of a > > 488 line? > > > > All the best, > > > > > > Tim > > > > Sent from my iPad > > > > On Nov 10, 2011, at 1:57 AM, Guy Cox <[hidden email]> wrote: > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > The Sapphire series lasers are OPS (Optically Pumped > > > DPSS but, you are right, they do indeed have a 514nm version. I don't > > > know why I didn't find it, but did find the 505, when I searched this > > > morning. I still think that the 505 might be exactly what Timothy > > > needs, though, since he doesn't want to buy an extra laser. > > > > > > I have also been contacted off-list and told that Spectra-Physics have > a > > > 515nm DPSS which is close enough to make no difference. I've no idea > > > what the lasing crystal is. > > > > > > Guy > > > > > > Optical Imaging Techniques in Cell Biology > > > by Guy Cox CRC Press / Taylor & Francis > > > http://www.guycox.com/optical.htm > > > ______________________________________________ > > > Associate Professor Guy Cox, MA, DPhil(Oxon) > > > Australian Centre for Microscopy & Microanalysis, > > > Madsen Building F09, University of Sydney, NSW 2006 > > > > > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > > Mobile 0413 281 861 > > > ______________________________________________ > > > http://www.guycox.net > > > > > > > > > > > > -----Original Message----- > > > From: Confocal Microscopy List [mailto: > [hidden email]] > > > On Behalf Of samuel connell > > > Sent: Thursday, 10 November 2011 1:57 PM > > > To: [hidden email] > > > Subject: Re: FRET in the time of DPSS > > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > Industry Response: > > > > > > There absolutely is a 514 DPSS available from 20 mW up to 150 mW > > > available > > > in the Sapphire form factor from Coherent. We use this laser, > > > conjunction with the 445 CUBE in our LaserStack for CFP/YFP (and their > > > newer brothers and sisters in FP evolution) imaging for FRET or > > > otherwise. > > > > > > ------------------------ > > > Samuel A. Connell > > > Sales Manager > > > Pacific Region-North America > > > Intelligent Imaging Innovations, Inc > > > 3250 Ocean Park Blvd, Suite 202 > > > Santa Monica, CA 90405 > > > Cell: (858) 692-4510 > > > [hidden email] > > > > > > > > > On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> > > > > > >> ***** > > >> To join, leave or search the confocal microscopy listserv, go to: > > >> > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > http://lists.umn > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > >> ***** > > >> > > >> See: http://www.cobolt.se/**coboltfandango515nm.html?** > > >> > > > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw< > http://www.cobolt.se/coboltfandango51 > > > 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> > > >> > > >> No connection, just considering about switching away from our old > > > Ion > > >> as well. > > >> > > >> - Damir > > >> > > >> > > >> On 11/9/2011 3:36 PM, Guy Cox wrote: > > >> > > >>> ***** > > >>> To join, leave or search the confocal microscopy listserv, go > > >>> > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > http://lists.umn > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > >>> ***** > > >>> > > >>> Are you sure there is a 514nm DPSS? I think that wavelength would > > > have > > >>> to be an OPS and I don't know anyone who makes one. Coherent do make > > > a > > >>> 505nm OPS which is intended to stand in for both the 488& 514 lines > > > of > > >>> > > >>> an Argon laser, and I rather suspect that this could be exactly what > > > you > > >>> need. But I have no experience of it. (I do have a Coherent 488nm > > >>> OPS). > > >>> > > >>> Guy > > >>> > > >>> Optical Imaging Techniques in Cell Biology > > >>> by Guy Cox CRC Press / Taylor& Francis > > >>> > > >>> > > > http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> > > >>> ______________________________**________________ > > >>> Associate Professor Guy Cox, MA, DPhil(Oxon) > > >>> Australian Centre for Microscopy& Microanalysis, > > >>> > > >>> Madsen Building F09, University of Sydney, NSW 2006 > > >>> > > >>> Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > >>> Mobile 0413 281 861 > > >>> ______________________________**________________ > > >>> http://www.guycox.net > > >>> > > >>> > > >>> > > >>> -----Original Message----- > > >>> From: Confocal Microscopy List > > > [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU > <[hidden email]. > > > EDU> > > >>> ] > > >>> On Behalf Of Tim Feinstein > > >>> Sent: Thursday, 10 November 2011 7:34 AM > > >>> To: [hidden email].**EDU > > > <[hidden email]> > > >>> Subject: FRET in the time of DPSS > > >>> > > >>> ***** > > >>> To join, leave or search the confocal microscopy listserv, go > > >>> > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > http://lists.umn > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > >>> ***** > > >>> > > >>> Hello all, > > >>> > > >>> We want to spec a four-laser launch for a new live cell system that > > > will > > >>> handle both CFP/YFP FRET and red/green imaging. However, I am sad > > > to > > >>> see that gas lasers are no longer speccable and so the freebie 514 > > > laser > > >>> line is gone. We would therefore have to spec a 514 DPSS and forego > > > the > > >>> far-red line. > > >>> > > >>> I was wondering whether there is a way to do more (or at least the > > > same) > > >>> with less. 488 nm excites YFP well enough, so in theory I could > > > image > > >>> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm > > > laser > > >>> lines. In my experience 442 nm laser excitation (via TIRF) causes > > >>> negligible YFP excitation and 488 nm does not excite CFP, so it is > > >>> possible that I could gain speed by passing everything through a > > > single > > >>> broad bandpass filter (e.g., 455-550 nm) and alternate excitations. > > >>> Assuming that cross-talk is not a problem, the most significant cost > > >>> would be that I lose a decent chunk of CFP emission to the scan head > > >>> dichroic, but in return I gain a 641 nm laser. > > >>> > > >>> Has anyone tried this? Any feedback on or off-list would be much > > >>> appreciated. > > >>> > > >>> Thanks and all the best, > > >>> > > >>> > > >>> TF > > >>> > > >>> Timothy Feinstein, PhD > > >>> Postdoctoral Fellow > > >>> Laboratory for GPCR Biology > > >>> Dept. of Pharmacology& Chemical Biology > > >>> > > >>> University of Pittsburgh, School of Medicine > > >>> BST W1301, 200 Lothrop St. > > >>> Pittsburgh, PA 15261 > > >>> > > >>> ----- > > >>> No virus found in this message. > > >>> Checked by AVG - www.avg.com > > >>> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: > > > 11/08/11 > > >>> > > >> > > >> -- > > >> Damir Sudar - Staff Scientist and Deputy for Technology > > >> Lawrence Berkeley Laboratory / Life Sciences Division > > >> One Cyclotron Road, MS 977, Berkeley, CA 94720, USA > > >> T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] > > >> WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** > > >> > > > Staff_Directory/Sudar_Lab.html< > http://www.lbl.gov/lsd/People_&_Organizat > > > ion/Scientific_Staff_Directory/Sudar_Lab.html> > > >> > > > > > > ----- > > > No virus found in this message. > > > Checked by AVG - www.avg.com > > > Version: 10.0.1411 / Virus Database: 2092/4006 - Release Date: > > > > |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** To add to this, the new 520nm diode lasers are of the first type Guy talks about. I agree that their center wavelengths can be slightly variable, but a good vendor will tell you exactly what wavelength is coming out of the laser you buy. Also, these diode lasers can be modulated electrically so you can use them without AOM devices. Craig On Fri, Nov 11, 2011 at 5:44 AM, Guy Cox <[hidden email]> wrote: > A simple diode laser is a pure semiconductor device. The laser cavity > is contained within a single chip (basically an LED with mirror layers > above and below it). Wavelengths currently available are from near UV > (~365nm) through violet to blue (~470nm), and then down in the red from > ~640nm. Because the laser cavity is so small the wavelength is not very > precise, which can be a problem for AOD based devices, though it is not > likely to be a biological problem. Your 405 and 440 nm lasers are most > likely simple diodes, as well and your 647nm. If you have a red laser > pointer this will be a simple diode. > > Correction from laser companies welcome, but I hope this helps. > > Guy > > > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) > Australian Centre for Microscopy & Microanalysis, > Madsen Building F09, University of Sydney, NSW 2006 > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Craig Brideau > Sent: Friday, 11 November 2011 12:05 PM > To: [hidden email] > Subject: Re: FRET in the time of DPSS > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Direct diode means that the laser has a single element. It tends to be > the > simplest type of laser; basically it is a big laser pointer. > Diode-pumped > solid state use an infrared diode (typically) which pumps some secondary > medium that gives you your desired wavelength. They are more complex > than > direct diode since you now have two separate objects that have to be > reliably optically coupled. Until recently direct diode lasers have > only > been capable of near-IR, Red, and Violet or Blue lines, but a recent > breakthrough in laser diode chemistry has allowed them to work at around > 520nm. > > Here's an old article from 2009 mentioning one of the first green direct > diode lasers: > > http://displaydaily.com/2009/07/21/green-laser-diode-demonstrated-a-brea > kthrough/ > > The current problem is pushing them to longer wavelengths. Projector > companies want to get them out to 530, 540nm for making displays and > video > projectors: > > http://www.qmed.com/mpmn/article/suppliers-chip-based-diode-makes-waves- > green-laser-world > > But 'shorter' direct diode green lasers in the 520nm or less range are > fairly available right now. > > Craig > > > > On Thu, Nov 10, 2011 at 5:09 PM, Jen Clarke > <[hidden email] > > wrote: > > > Hi all > > > > This discussion is very timely for our facility, as we are looking to > > purchase a solide state laser in the ~500-520nm range as soon as > possible > > > > I am insufficiently familiar with the pros and cons of choosing a DPSS > > versus a OPS or a direct-diode laser > > > > From the discussion so far it sounds like the choices for a solid > state > > laser for imaging YFP are 515nm DPSS from Spectra-Physics, a 505nm > Sapphire > > OPS, a 514nm Sapphire OPS or a 520 direct diode. > > > > Guy - I presume by "direct diode" you mean a diode pumped solid state, > > which I understand would be not as good as a DPSS (please correct me > if I > > am wrong) > > > > I dont have any idea how an OPS compares to a DPSS laser > > > > Any advice as to which of these options might be "best" as an add on > to a > > new Olympus FV1000 for the primary purpose of imaging and bleaching > YFP in > > the CFP/YFP FRET pair would be greatly appreciated. > > > > Kind regards > > Jen > > -- > > Jennifer Clarke BSc (Hons) PhD > > Research Associate, Anatomy and Histology > > Centre for Neuroscience, School of Medicine > > & > > Facility Manager, Optical Microscopy Suite, Flinders Microscopy > > > > Flinders University > > GPO Box 2100, Adelaide 5001 > > Phone: 61 8 8204 6454/ 61 8 8204 6637 > > Email: [hidden email] > > > > > > ________________________________________ > > From: Confocal Microscopy List [[hidden email]] On > > Behalf Of Craig Brideau [[hidden email]] > > Sent: Friday, 11 November 2011 5:03 AM > > To: [hidden email] > > Subject: Re: FRET in the time of DPSS > > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > There are also the new direct diode lasers at 520nm if you can handle > a 6nm > > red wavelength shift from 514. They've only been around for about a > year > > but many manufacturers have lumped them in with their direct-diode > product > > lines and they look pretty decent. > > > > Craig > > > > > > > > On Thu, Nov 10, 2011 at 6:08 AM, Tim Feinstein <[hidden email]> wrote: > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > Hi guys (and Guy), > > > > > > Many thanks for the feedback. I like the 505 laser idea in > particular. > > > As long as it excites 488 fluorophores well enough it could be > exactly > > > what I need. Does anybody have experience with using that in place > of a > > > 488 line? > > > > > > All the best, > > > > > > > > > Tim > > > > > > Sent from my iPad > > > > > > On Nov 10, 2011, at 1:57 AM, Guy Cox <[hidden email]> wrote: > > > > > > > ***** > > > > To join, leave or search the confocal microscopy listserv, go to: > > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > > ***** > > > > > > > > The Sapphire series lasers are OPS (Optically Pumped > Semiconductor) not > > > > DPSS but, you are right, they do indeed have a 514nm version. I > don't > > > > know why I didn't find it, but did find the 505, when I searched > this > > > > morning. I still think that the 505 might be exactly what Timothy > > > > needs, though, since he doesn't want to buy an extra laser. > > > > > > > > I have also been contacted off-list and told that Spectra-Physics > have > > a > > > > 515nm DPSS which is close enough to make no difference. I've no > idea > > > > what the lasing crystal is. > > > > > > > > Guy > > > > > > > > Optical Imaging Techniques in Cell Biology > > > > by Guy Cox CRC Press / Taylor & Francis > > > > http://www.guycox.com/optical.htm > > > > ______________________________________________ > > > > Associate Professor Guy Cox, MA, DPhil(Oxon) > > > > Australian Centre for Microscopy & Microanalysis, > > > > Madsen Building F09, University of Sydney, NSW 2006 > > > > > > > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > > > Mobile 0413 281 861 > > > > ______________________________________________ > > > > http://www.guycox.net > > > > > > > > > > > > > > > > -----Original Message----- > > > > From: Confocal Microscopy List [mailto: > > [hidden email]] > > > > On Behalf Of samuel connell > > > > Sent: Thursday, 10 November 2011 1:57 PM > > > > To: [hidden email] > > > > Subject: Re: FRET in the time of DPSS > > > > > > > > ***** > > > > To join, leave or search the confocal microscopy listserv, go to: > > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > > ***** > > > > > > > > Industry Response: > > > > > > > > There absolutely is a 514 DPSS available from 20 mW up to 150 mW > > > > available > > > > in the Sapphire form factor from Coherent. We use this laser, > often in > > > > conjunction with the 445 CUBE in our LaserStack for CFP/YFP (and > their > > > > newer brothers and sisters in FP evolution) imaging for FRET or > > > > otherwise. > > > > > > > > ------------------------ > > > > Samuel A. Connell > > > > Sales Manager > > > > Pacific Region-North America > > > > Intelligent Imaging Innovations, Inc > > > > 3250 Ocean Park Blvd, Suite 202 > > > > Santa Monica, CA 90405 > > > > Cell: (858) 692-4510 > > > > [hidden email] > > > > > > > > > > > > On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> > wrote: > > > > > > > >> ***** > > > >> To join, leave or search the confocal microscopy listserv, go to: > > > >> > > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > > http://lists.umn > > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > > >> ***** > > > >> > > > >> See: http://www.cobolt.se/**coboltfandango515nm.html?** > > > >> > > > > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw< > > http://www.cobolt.se/coboltfandango51 > > > > 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> > > > >> > > > >> No connection, just considering about switching away from our old > Ar > > > > Ion > > > >> as well. > > > >> > > > >> - Damir > > > >> > > > >> > > > >> On 11/9/2011 3:36 PM, Guy Cox wrote: > > > >> > > > >>> ***** > > > >>> To join, leave or search the confocal microscopy listserv, go > to: > > > >>> > > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > > http://lists.umn > > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > > >>> ***** > > > >>> > > > >>> Are you sure there is a 514nm DPSS? I think that wavelength > would > > > > have > > > >>> to be an OPS and I don't know anyone who makes one. Coherent do > make > > > > a > > > >>> 505nm OPS which is intended to stand in for both the 488& 514 > lines > > > > of > > > >>> > > > >>> an Argon laser, and I rather suspect that this could be exactly > what > > > > you > > > >>> need. But I have no experience of it. (I do have a Coherent > 488nm > > > >>> OPS). > > > >>> > > > >>> Guy > > > >>> > > > >>> Optical Imaging Techniques in Cell Biology > > > >>> by Guy Cox CRC Press / Taylor& Francis > > > >>> > > > >>> > > > > > http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> > > > >>> ______________________________**________________ > > > >>> Associate Professor Guy Cox, MA, DPhil(Oxon) > > > >>> Australian Centre for Microscopy& Microanalysis, > > > >>> > > > >>> Madsen Building F09, University of Sydney, NSW 2006 > > > >>> > > > >>> Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > > >>> Mobile 0413 281 861 > > > >>> ______________________________**________________ > > > >>> http://www.guycox.net > > > >>> > > > >>> > > > >>> > > > >>> -----Original Message----- > > > >>> From: Confocal Microscopy List > > > > [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU > > <[hidden email]. > > > > EDU> > > > >>> ] > > > >>> On Behalf Of Tim Feinstein > > > >>> Sent: Thursday, 10 November 2011 7:34 AM > > > >>> To: [hidden email].**EDU > > > > <[hidden email]> > > > >>> Subject: FRET in the time of DPSS > > > >>> > > > >>> ***** > > > >>> To join, leave or search the confocal microscopy listserv, go > to: > > > >>> > > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > > http://lists.umn > > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > > >>> ***** > > > >>> > > > >>> Hello all, > > > >>> > > > >>> We want to spec a four-laser launch for a new live cell system > that > > > > will > > > >>> handle both CFP/YFP FRET and red/green imaging. However, I am > sad > > > > to > > > >>> see that gas lasers are no longer speccable and so the freebie > 514 > > > > laser > > > >>> line is gone. We would therefore have to spec a 514 DPSS and > forego > > > > the > > > >>> far-red line. > > > >>> > > > >>> I was wondering whether there is a way to do more (or at least > the > > > > same) > > > >>> with less. 488 nm excites YFP well enough, so in theory I could > > > > image > > > >>> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 > nm > > > > laser > > > >>> lines. In my experience 442 nm laser excitation (via TIRF) > causes > > > >>> negligible YFP excitation and 488 nm does not excite CFP, so it > is > > > >>> possible that I could gain speed by passing everything through a > > > > single > > > >>> broad bandpass filter (e.g., 455-550 nm) and alternate > excitations. > > > >>> Assuming that cross-talk is not a problem, the most significant > cost > > > >>> would be that I lose a decent chunk of CFP emission to the scan > head > > > >>> dichroic, but in return I gain a 641 nm laser. > > > >>> > > > >>> Has anyone tried this? Any feedback on or off-list would be > much > > > >>> appreciated. > > > >>> > > > >>> Thanks and all the best, > > > >>> > > > >>> > > > >>> TF > > > >>> > > > >>> Timothy Feinstein, PhD > > > >>> Postdoctoral Fellow > > > >>> Laboratory for GPCR Biology > > > >>> Dept. of Pharmacology& Chemical Biology > > > >>> > > > >>> University of Pittsburgh, School of Medicine > > > >>> BST W1301, 200 Lothrop St. > > > >>> Pittsburgh, PA 15261 > > > >>> > > > >>> ----- > > > >>> No virus found in this message. > > > >>> Checked by AVG - www.avg.com > > > >>> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: > > > > 11/08/11 > > > >>> > > > >> > > > >> -- > > > >> Damir Sudar - Staff Scientist and Deputy for Technology > > > >> Lawrence Berkeley Laboratory / Life Sciences Division > > > >> One Cyclotron Road, MS 977, Berkeley, CA 94720, USA > > > >> T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] > > > >> WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** > > > >> > > > > Staff_Directory/Sudar_Lab.html< > > http://www.lbl.gov/lsd/People_&_Organizat > > > > ion/Scientific_Staff_Directory/Sudar_Lab.html> > > > >> > > > > > > > > ----- > > > > No virus found in this message. > > > > Checked by AVG - www.avg.com > > > > Version: 10.0.1411 / Virus Database: 2092/4006 - Release Date: > 11/09/11 > > > > > > > > |
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Arrigoni, Marco |
Re: FRET in the time of DPSS - commercial posting
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In reply to this post by Guy Cox-2
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** As Guy correctly mentioned, the OPS technology is unique to Coherent and uses a diode-pumped semiconductor chip to generate infrared radiation that is then intra-cavity doubled or tripled to produce CW visible/UV light. This technology is flexible in wavelength and highly scalable in power. For example, Coherent offers OPS devices with wavelengths in the range 460-589 nm and power levels ranging from milliwatts to 10 watts. Frequency-tripled versions operate at 355 nm. A number of products using OPS technology are on display this week at Neuroscience. Details of these products and the OPS technology can be found at http://www.coherent.com/products/?771/Optically-Pumped-Semiconductor-Laser-OPSL-Technology Best regards, Marco Arrigoni Director of Marketing Scientific Market Segment Coherent, Inc. 408-764-4661 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: Friday, November 11, 2011 4:45 AM To: [hidden email] Subject: Re: FRET in the time of DPSS ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** OK, this is quite a confusing area (and the manufacturers do not always make things as clear as they could). A simple diode laser is a pure semiconductor device. The laser cavity is contained within a single chip (basically an LED with mirror layers above and below it). Wavelengths currently available are from near UV (~365nm) through violet to blue (~470nm), and then down in the red from ~640nm. Because the laser cavity is so small the wavelength is not very precise, which can be a problem for AOD based devices, though it is not likely to be a biological problem. Your 405 and 440 nm lasers are most likely simple diodes, as well and your 647nm. If you have a red laser pointer this will be a simple diode. OPS (optically pumped semiconductor) lasers are semiconductor devices which are not excited by electric current but by input from a diode laser. They can access wavelengths which direct diodes cannot. I think it is fair to say that this has been very much Coherent technology. 488 - 515 lasers, which are tricky from other sources, are within this range. DPSS are solid state crystal lasers pumped by red diode lasers. Neodymium Yttrium Aluminium Garnet (NdYAG) is the classic, working at 1064nm. For our purposes we are mostly interested in lasers which are frequency doubled by second harmonic generating crystals and the classic is therefore 532nm - the wavelength of your green laser pointer and the laser which pumps your TiS laser for multiphoton microscopy. Ytterbium lasers give 1030 nm (and hence 515nm when doubled). 561 nm is another option, and I believe it comes from a secondary line from an Nd YAG. Correction from laser companies welcome, but I hope this helps. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Australian Centre for Microscopy & Microanalysis, Madsen Building F09, University of Sydney, NSW 2006 Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Craig Brideau Sent: Friday, 11 November 2011 12:05 PM To: [hidden email] Subject: Re: FRET in the time of DPSS ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Direct diode means that the laser has a single element. It tends to be the simplest type of laser; basically it is a big laser pointer. Diode-pumped solid state use an infrared diode (typically) which pumps some secondary medium that gives you your desired wavelength. They are more complex than direct diode since you now have two separate objects that have to be reliably optically coupled. Until recently direct diode lasers have only been capable of near-IR, Red, and Violet or Blue lines, but a recent breakthrough in laser diode chemistry has allowed them to work at around 520nm. Here's an old article from 2009 mentioning one of the first green direct diode lasers: http://displaydaily.com/2009/07/21/green-laser-diode-demonstrated-a-brea kthrough/ The current problem is pushing them to longer wavelengths. Projector companies want to get them out to 530, 540nm for making displays and video projectors: http://www.qmed.com/mpmn/article/suppliers-chip-based-diode-makes-waves- green-laser-world But 'shorter' direct diode green lasers in the 520nm or less range are fairly available right now. Craig On Thu, Nov 10, 2011 at 5:09 PM, Jen Clarke <[hidden email] > wrote: > Hi all > > This discussion is very timely for our facility, as we are looking to > purchase a solide state laser in the ~500-520nm range as soon as possible > > I am insufficiently familiar with the pros and cons of choosing a DPSS > versus a OPS or a direct-diode laser > > From the discussion so far it sounds like the choices for a solid state > laser for imaging YFP are 515nm DPSS from Spectra-Physics, a 505nm Sapphire > OPS, a 514nm Sapphire OPS or a 520 direct diode. > > Guy - I presume by "direct diode" you mean a diode pumped solid state, > which I understand would be not as good as a DPSS (please correct me if I > am wrong) > > I dont have any idea how an OPS compares to a DPSS laser > > Any advice as to which of these options might be "best" as an add on to a > new Olympus FV1000 for the primary purpose of imaging and bleaching YFP in > the CFP/YFP FRET pair would be greatly appreciated. > > Kind regards > Jen > -- > Jennifer Clarke BSc (Hons) PhD > Research Associate, Anatomy and Histology > Centre for Neuroscience, School of Medicine > & > Facility Manager, Optical Microscopy Suite, Flinders Microscopy > > Flinders University > GPO Box 2100, Adelaide 5001 > Phone: 61 8 8204 6454/ 61 8 8204 6637 > Email: [hidden email] > > > ________________________________________ > From: Confocal Microscopy List [[hidden email]] On > Behalf Of Craig Brideau [[hidden email]] > Sent: Friday, 11 November 2011 5:03 AM > To: [hidden email] > Subject: Re: FRET in the time of DPSS > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > There are also the new direct diode lasers at 520nm if you can handle > red wavelength shift from 514. They've only been around for about a year > but many manufacturers have lumped them in with their direct-diode product > lines and they look pretty decent. > > Craig > > > > On Thu, Nov 10, 2011 at 6:08 AM, Tim Feinstein <[hidden email]> wrote: > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Hi guys (and Guy), > > > > Many thanks for the feedback. I like the 505 laser idea in > > As long as it excites 488 fluorophores well enough it could be exactly > > what I need. Does anybody have experience with using that in place of a > > 488 line? > > > > All the best, > > > > > > Tim > > > > Sent from my iPad > > > > On Nov 10, 2011, at 1:57 AM, Guy Cox <[hidden email]> wrote: > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > The Sapphire series lasers are OPS (Optically Pumped > > > DPSS but, you are right, they do indeed have a 514nm version. I don't > > > know why I didn't find it, but did find the 505, when I searched this > > > morning. I still think that the 505 might be exactly what Timothy > > > needs, though, since he doesn't want to buy an extra laser. > > > > > > I have also been contacted off-list and told that Spectra-Physics have > a > > > 515nm DPSS which is close enough to make no difference. I've no idea > > > what the lasing crystal is. > > > > > > Guy > > > > > > Optical Imaging Techniques in Cell Biology > > > by Guy Cox CRC Press / Taylor & Francis > > > http://www.guycox.com/optical.htm > > > ______________________________________________ > > > Associate Professor Guy Cox, MA, DPhil(Oxon) > > > Australian Centre for Microscopy & Microanalysis, > > > Madsen Building F09, University of Sydney, NSW 2006 > > > > > > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > > Mobile 0413 281 861 > > > ______________________________________________ > > > http://www.guycox.net > > > > > > > > > > > > -----Original Message----- > > > From: Confocal Microscopy List [mailto: > [hidden email]] > > > On Behalf Of samuel connell > > > Sent: Thursday, 10 November 2011 1:57 PM > > > To: [hidden email] > > > Subject: Re: FRET in the time of DPSS > > > > > > ***** > > > To join, leave or search the confocal microscopy listserv, go to: > > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > > ***** > > > > > > Industry Response: > > > > > > There absolutely is a 514 DPSS available from 20 mW up to 150 mW > > > available > > > in the Sapphire form factor from Coherent. We use this laser, > > > conjunction with the 445 CUBE in our LaserStack for CFP/YFP (and their > > > newer brothers and sisters in FP evolution) imaging for FRET or > > > otherwise. > > > > > > ------------------------ > > > Samuel A. Connell > > > Sales Manager > > > Pacific Region-North America > > > Intelligent Imaging Innovations, Inc > > > 3250 Ocean Park Blvd, Suite 202 > > > Santa Monica, CA 90405 > > > Cell: (858) 692-4510 > > > [hidden email] > > > > > > > > > On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[hidden email]> > > > > > >> ***** > > >> To join, leave or search the confocal microscopy listserv, go to: > > >> > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > http://lists.umn > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > >> ***** > > >> > > >> See: http://www.cobolt.se/**coboltfandango515nm.html?** > > >> > > > gclid=CPCOueXlqqwCFQyEhwodQn-**aAw< > http://www.cobolt.se/coboltfandango51 > > > 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw> > > >> > > >> No connection, just considering about switching away from our old > > > Ion > > >> as well. > > >> > > >> - Damir > > >> > > >> > > >> On 11/9/2011 3:36 PM, Guy Cox wrote: > > >> > > >>> ***** > > >>> To join, leave or search the confocal microscopy listserv, go > > >>> > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > http://lists.umn > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > >>> ***** > > >>> > > >>> Are you sure there is a 514nm DPSS? I think that wavelength would > > > have > > >>> to be an OPS and I don't know anyone who makes one. Coherent do make > > > a > > >>> 505nm OPS which is intended to stand in for both the 488& 514 lines > > > of > > >>> > > >>> an Argon laser, and I rather suspect that this could be exactly what > > > you > > >>> need. But I have no experience of it. (I do have a Coherent 488nm > > >>> OPS). > > >>> > > >>> Guy > > >>> > > >>> Optical Imaging Techniques in Cell Biology > > >>> by Guy Cox CRC Press / Taylor& Francis > > >>> > > >>> > > > http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm> > > >>> ______________________________**________________ > > >>> Associate Professor Guy Cox, MA, DPhil(Oxon) > > >>> Australian Centre for Microscopy& Microanalysis, > > >>> > > >>> Madsen Building F09, University of Sydney, NSW 2006 > > >>> > > >>> Phone +61 2 9351 3176 Fax +61 2 9351 7682 > > >>> Mobile 0413 281 861 > > >>> ______________________________**________________ > > >>> http://www.guycox.net > > >>> > > >>> > > >>> > > >>> -----Original Message----- > > >>> From: Confocal Microscopy List > > > [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU > <[hidden email]. > > > EDU> > > >>> ] > > >>> On Behalf Of Tim Feinstein > > >>> Sent: Thursday, 10 November 2011 7:34 AM > > >>> To: [hidden email].**EDU > > > <[hidden email]> > > >>> Subject: FRET in the time of DPSS > > >>> > > >>> ***** > > >>> To join, leave or search the confocal microscopy listserv, go > > >>> > > > http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy< > http://lists.umn > > > .edu/cgi-bin/wa?A0=confocalmicroscopy> > > >>> ***** > > >>> > > >>> Hello all, > > >>> > > >>> We want to spec a four-laser launch for a new live cell system that > > > will > > >>> handle both CFP/YFP FRET and red/green imaging. However, I am sad > > > to > > >>> see that gas lasers are no longer speccable and so the freebie 514 > > > laser > > >>> line is gone. We would therefore have to spec a 514 DPSS and forego > > > the > > >>> far-red line. > > >>> > > >>> I was wondering whether there is a way to do more (or at least the > > > same) > > >>> with less. 488 nm excites YFP well enough, so in theory I could > > > image > > >>> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm > > > laser > > >>> lines. In my experience 442 nm laser excitation (via TIRF) causes > > >>> negligible YFP excitation and 488 nm does not excite CFP, so it is > > >>> possible that I could gain speed by passing everything through a > > > single > > >>> broad bandpass filter (e.g., 455-550 nm) and alternate excitations. > > >>> Assuming that cross-talk is not a problem, the most significant cost > > >>> would be that I lose a decent chunk of CFP emission to the scan head > > >>> dichroic, but in return I gain a 641 nm laser. > > >>> > > >>> Has anyone tried this? Any feedback on or off-list would be much > > >>> appreciated. > > >>> > > >>> Thanks and all the best, > > >>> > > >>> > > >>> TF > > >>> > > >>> Timothy Feinstein, PhD > > >>> Postdoctoral Fellow > > >>> Laboratory for GPCR Biology > > >>> Dept. of Pharmacology& Chemical Biology > > >>> > > >>> University of Pittsburgh, School of Medicine > > >>> BST W1301, 200 Lothrop St. > > >>> Pittsburgh, PA 15261 > > >>> > > >>> ----- > > >>> No virus found in this message. > > >>> Checked by AVG - www.avg.com > > >>> Version: 10.0.1411 / Virus Database: 2092/4005 - Release Date: > > > 11/08/11 > > >>> > > >> > > >> -- > > >> Damir Sudar - Staff Scientist and Deputy for Technology > > >> Lawrence Berkeley Laboratory / Life Sciences Division > > >> One Cyclotron Road, MS 977, Berkeley, CA 94720, USA > > >> T: 510/486-5346 - F: 510/486-5586 - E: [hidden email] > > >> WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_** > > >> > > > Staff_Directory/Sudar_Lab.html< > http://www.lbl.gov/lsd/People_&_Organizat > > > ion/Scientific_Staff_Directory/Sudar_Lab.html> > > >> > > > > > > ----- > > > No virus found in this message. > > > Checked by AVG - www.avg.com > > > Version: 10.0.1411 / Virus Database: 2092/4006 - Release Date: > > > > The information contained in this communication is confidential and may be legally privileged. It is intended solely for the use of the individual or entity to whom it is addressed and others authorized to receive it. If you are not the intended recipient you are hereby (a) notified that any disclosure, copying, distribution or taking any action with respect to the content of this information is strictly prohibited and may be unlawful, and (b) kindly requested to inform the sender immediately and delete any copies. Coherent is neither liable for the proper and complete transmission of the information contained in this communication nor for any delay in its receipt. |
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James Pawley |
Re: Deconvolution of Confocal Images? (was: Airy Units)
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In reply to this post by James Mansfield
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Jim Mansfield makes a good point about "clutter" being a problem in automatic image analysis. However, when I talk about "the noise produced by out-of-focus light" I am really talking about noise. I do not refer to the intensity of the signal caused by this out-of-focus light but merely the Poisson Noise associated with this signal. Given a large, bright blob located away from the focus plane, this Poison Noise can swamp the "real" signal from a small in-focus feature. JP > >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >Hi, > >I'm not sure I like calling out-of-focal-plane photons "noise". They >are not noise, they are just signals from places where you don't >happen to want them at the moment. > >To take a set of terms from another field, the military >hyperspectral imaging crowd (who are generally trying to isolate the >signal from a tank or other military object from the background of >normal non-military objects) have come up with: > >Noise: random or instrument signals that are an artifact of the way >you collected the data >Signal: the signal you actually care about >Clutter: other (real) signals that interfering with your ability to >see the signal > >Depending on the sample, either "noise" or "clutter" are the >limiting factors. For many samples looked at in confocal, noise is >the limiting factor. For many highly autofluorescent tissue samples, >clutter is the limiting factor and needs to be dealt with >differently than random noise. >Mike Rafferty <[hidden email]> >Jim > > > >James R. Mansfield >Director, Tissue Analysis Applications >Caliper Life Sciences, Inc. >68 Elm Street, Hopkinton, MA 01748 >Office: +1 774 278 2802 >Cell: +1 617 416 6175 >Email: [hidden email] >www.caliperls.com >www.cri-inc.com > >Need to send me large files? Use my YouSendIt DropBox: >https://dropbox.yousendit.com/Mansfield-Dropbox > > >-----Original Message----- >From: Confocal Microscopy List >[mailto:[hidden email]] On Behalf Of Guy Cox >Sent: Monday, October 31, 2011 7:21 AM >To: [hidden email] >Subject: Re: Deconvolution of Confocal Images? (was: Airy Units) > >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >Daniel White wrote: > >" in a confocal you throw away most of the signal, as its out of focus. >So as a result the images are often very noisy. " > >This is often stated but IT IS TOTALLY UNTRUE. What is out of focus is >noise, not signal. If you have no SA (and, honestly, if you are >seriously interested in high-resolution imaging that should be a given) >then a confocal microscope with the pinhole set at 1 Airy diameter >throws away no signal at all. So why are confocal images often noisy? >Well, it's just statistics. If you take a wide-field image with a 1 >second exposure each point is exposed for one second. If you take a >confocal image at 512 x 512 for 1 second then each point is exposed for >~4 microseconds. The difference is rather substantial ... > > Guy > >Optical Imaging Techniques in Cell Biology >by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm >______________________________________________ >Associate Professor Guy Cox, MA, DPhil(Oxon) >Australian Centre for Microscopy & Microanalysis, >Madsen Building F09, University of Sydney, NSW 2006 > >Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 >______________________________________________ > http://www.guycox.net > > >-----Original Message----- >From: Confocal Microscopy List [mailto:[hidden email]] >On Behalf Of daniel white >Sent: Monday, 31 October 2011 8:30 PM >To: [hidden email] >Subject: Deconvolution of Confocal Images? (was: Airy Units) > >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >Hi Peter, > >On Oct 31, 2011, at 6:02 AM, CONFOCALMICROSCOPY automatic digest system >wrote: > >> >> Date: Sun, 30 Oct 2011 13:09:10 -0700 >> From: Peter Werner <[hidden email]> >> Subject: Deconvolution of Confocal Images? (was: Airy Units) >> >> An interesting point was made here by Jim Pawley: >> >>> I agree that sampling a bit higher than Nyquist never hurts, >>> especially if you deconvolve (as you always should), but I think >>> that it is a mistake to think that one can "separate" out the noise >>> by decon. I think that noise is pretty fundamental. >> >> I had always heard that if you're doing confocal microscopy, at least > >> point-scanning confocal with a pinhole size of 1AU or smaller, that >> deconvolution was superfluous, because you shouldn't be getting out of > >> focus light. So what is gained by deconvolution when one is sampling >> voxel by voxel? > >in a confocal you throw away most of the signal, as its out of focus. >So as a result the images are often very noisy. >Good contrast.... but high Poisson distributed photon shot noise >from only measuring a handful of photons. > >So usually one needs to do something about that noise... >we want to separate the real signal from the noise. > >Often a Gaussian or mean filter is applied... which suppresses the noise > >by smoothing it out... but it also smooths the real signal, so >effectively you lose >the contrast and resolution that was the whole point of doing confocal. > >The smart way to suppress the noise, but keep the contrast and >resolution >is to do deconvolution. >Deconvolution using a max likelyhood method uses the known shape of the >PSF >to make a best guess model of the real fluorophore distribution in the >sample. >You tell the deconvolution algorithm how noisy the image is (you have to >guess >unless you take 2 images and measure it) >then it attempts to throw out the noise and keep the real signal, >resolution and contrast intact. > >D > >> >> Peter G. Werner >> Merritt College Microscopy Program > >Dr. Daniel James White BSc. (Hons.) PhD > >Leader - Image Processing Facility, >Senior Microscopist, >Light Microscopy Facility. > >Max Planck Institute of Molecular Cell Biology and Genetics >Pfotenhauerstrasse 108 >01307 DRESDEN >Germany > >+49 (0)15114966933 (German Mobile) >+49 (0)351 210 2627 (Work phone at MPI-CBG) >+49 (0)351 210 1078 (Fax MPI-CBG LMF) >chalkie666 Skype >http://www.bioimagexd.net BioImageXD >http://fiji.sc Fiji - is just ImageJ >(Batteries Included) >http://www.chalkie.org.uk Dan's Homepages >https://ifn.mpi-cbg.de Biopolis Dresden Imaging >Platform (BioDIP) >dan (at) chalkie.org.uk >( white (at) mpi-cbg.de ) > >----- >No virus found in this message. >Checked by AVG - www.avg.com >Version: 10.0.1411 / Virus Database: 2092/3985 - Release Date: 10/30/11 > >============================================================= The >information contained in this e-mail message is intended only for >the personal and confidential use of the recipient(s) named above. >If the reader of this message is not the intended recipient or an >agent responsible for delivering it to the intended recipient, you >are hereby notified that you have received this document in error >and that any review, dissemination, distribution, or copying of this >message is strictly prohibited. If you have received this >communication in error, please notify us immediately by e-mail, and >delete the original message. >============================================================= > >==================================================================================== > >The information contained in this e-mail message is intended only >for the personal and confidential use of the recipient(s) named >above. If the reader of this message is not the intended recipient >or an agent responsible for delivering it to the intended recipient, >you are hereby notified that you have received this document in >error and that any review, dissemination, distribution, or copying >of this message is strictly prohibited. If you have received this >communication in error, please notify us immediately by e-mail, and >delete the original message. > >==================================================================================== -- James and Christine Pawley, 21 N. Prospect Ave. Madison, WI, 53726 Phone: 608-238-3953 |
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Tim O'Brien Sr. |
3D viewing of your stacks at ASCB - Booth 853
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear Confocal Listserve users- If you have stacks of cells stained for tubular structures (or filaments) plus another fluorophore, and are attending ASCB, we would like to view them at the meeting in Denver. We are trying to improve our free 3D stereo rendering program (ImageSurfer 2.0) and make it a seriously useful program for Cell Biologists. With inexpensive 3D viewing systems now easily assembled (for less than $5000 including computer and monitor, glasses, etc), we feel ImageSurfer could be a great way to routinely view collocalizations within cells in many labs. The new functionality within ISurfer allows much better rendering of tubular structures (a subject dear to my heart), plus rendering of multiple colors should make it a useful tool for visualizing and quantifying relative locations of proteins within cells. The program is freely available at our website (http://cismm.cs.unc.edu/) along with many other useful programs such as the microscope simulator, image tracker, and so on. We also have a how to for assembling your own stereo viewing system for your lab. The advantage to viewing your stacks in Denver is that we can learn what you like and how we can improve it. You can view your images with 3D glasses at our booth, and take back a movie of the rendering. If you aren't going to ASCB, please feel free to download the program and try it out. Let me know what can be improved, or contact us through the website. If you are going, please consider bringing a thumb drive with your stacks to booth 853. Thanks! Tim O'Brien -- E. Tim O'Brien Sr. Computer Integrated Systems for Microscopy and Manipulation UNC Physics and Astronomy UNC Chapel Hill, NC 27599 919 843 2798 |
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Andreas Bruckbauer |
Re: 3D viewing of your stacks at ASCB - Booth 853
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi, that sounds great, viewing our nice 3D and 4D data with up to date technology. I heard Imaris can do the same, does anyone have experience with this? I would be interested in the specs for the graphic cards for Imaris. best wishes Andreas -----Original Message----- From: Tim O'Brien Sr. <[hidden email]> To: CONFOCALMICROSCOPY <[hidden email]> Sent: Tue, 29 Nov 2011 22:57 Subject: 3D viewing of your stacks at ASCB - Booth 853 ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear Confocal Listserve users- If you have stacks of cells stained for tubular structures (or filaments) plus another fluorophore, and are attending ASCB, we would like to view them at the meeting in Denver. We are trying to improve our free 3D stereo rendering program (ImageSurfer 2.0) and make it a seriously useful program for Cell Biologists. With inexpensive 3D viewing systems now easily assembled (for less than $5000 including computer and monitor, glasses, etc), we feel ImageSurfer could be a great way to routinely view collocalizations within cells in many labs. The new functionality within ISurfer allows much better rendering of tubular structures (a subject dear to my heart), plus rendering of multiple colors should make it a useful tool for visualizing and quantifying relative locations of proteins within cells. The program is freely available at our website (http://cismm.cs.unc.edu/) along with many other useful programs such as the microscope simulator, image tracker, and so on. We also have a how to for assembling your own stereo viewing system for your lab. The advantage to viewing your stacks in Denver is that we can learn what you like and how we can improve it. You can view your images with 3D glasses at our booth, and take back a movie of the rendering. If you aren't going to ASCB, please feel free to download the program and try it out. Let me know what can be improved, or contact us through the website. If you are going, please consider bringing a thumb drive with your stacks to booth 853. Thanks! Tim O'Brien -- E. Tim O'Brien Sr. Computer Integrated Systems for Microscopy and Manipulation UNC Physics and Astronomy UNC Chapel Hill, NC 27599 919 843 2798 |
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Tim O'Brien Sr. |
Re: 3D viewing of your stacks at ASCB - Booth 853
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Andreas- Our software is up to date, especially in the sense that it is still evolving. Definitely usable and useful, and we want help prioritizing the next steps/features needed in the new version. See you soon, Tim On 12/1/2011 4:10 PM, Andreas Bruckbauer wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi, > that sounds great, viewing our nice 3D and 4D data with up to date technology. I heard Imaris can do the same, does anyone have experience with this? I would be interested in the specs for the graphic cards for Imaris. > > best wishes > > Andreas > > > > > > > > > -----Original Message----- > From: Tim O'Brien Sr.<[hidden email]> > To: CONFOCALMICROSCOPY<[hidden email]> > Sent: Tue, 29 Nov 2011 22:57 > Subject: 3D viewing of your stacks at ASCB - Booth 853 > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Dear Confocal Listserve users- > > > > If you have stacks of cells stained for tubular structures (or > > filaments) plus another fluorophore, and are attending ASCB, we would > > like to view them at the meeting in Denver. We are trying to improve > > our free 3D stereo rendering program (ImageSurfer 2.0) and make it a > > seriously useful program for Cell Biologists. > > > > With inexpensive 3D viewing systems now easily assembled (for less than > > $5000 including computer and monitor, glasses, etc), we feel ImageSurfer > > could be a great way to routinely view collocalizations within cells in > > many labs. The new functionality within ISurfer allows much better > > rendering of tubular structures (a subject dear to my heart), plus > > rendering of multiple colors should make it a useful tool for > > visualizing and quantifying relative locations of proteins within > > cells. The program is freely available at our website > > (http://cismm.cs.unc.edu/) along with many other useful programs such as > > the microscope simulator, image tracker, and so on. We also have a how > > to for assembling your own stereo viewing system for your lab. > > > > The advantage to viewing your stacks in Denver is that we can learn what > > you like and how we can improve it. You can view your images with 3D > > glasses at our booth, and take back a movie of the rendering. > > > > If you aren't going to ASCB, please feel free to download the program > > and try it out. Let me know what can be improved, or contact us through > > the website. If you are going, please consider bringing a thumb drive > > with your stacks to booth 853. > > > > Thanks! > > > > Tim O'Brien > > > -- E. Tim O'Brien Sr. Computer Integrated Systems for Microscopy and Manipulation UNC Physics and Astronomy UNC Chapel Hill, NC 27599 919 843 2798 |
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George McNamara |
Zeiss released ZEN Lite 2011 with nice 3D rendering ... Re: 3D viewing of your stacks at ASCB - Booth 853
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Tim, Thanks for letting us know about your new iSurfer 2.0. I realize it is free, but can't resist the question I always ask about software: Does it have a working Print money button? Not 3D stereo, but the Zeiss ZEN Lite 2011 available at http://www.zeiss.de/C12567BE0045ACF1/Contents-Frame/72E7F802F859568FC125734C0039B812 has nice 3D rendering. Hopefully other image acquisition oriented companies will enable some - or all - their cool features in their free viewing/analysis software. Sincerely, George p.s. My thanks to my UM colleague Justin Price for the ZEN URL. Justin was the Zeiss summer intern at MBL and CSHL this past summer. On 12/1/2011 5:36 PM, Tim O'Brien Sr. wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Andreas- > > Our software is up to date, especially in the sense that it is still > evolving. Definitely usable and useful, and we want help prioritizing > the next steps/features needed in the new version. > > See you soon, > Tim > > > On 12/1/2011 4:10 PM, Andreas Bruckbauer wrote: >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Hi, >> that sounds great, viewing our nice 3D and 4D data with up to date >> technology. I heard Imaris can do the same, does anyone have >> experience with this? I would be interested in the specs for the >> graphic cards for Imaris. >> >> best wishes >> >> Andreas >> >> >> >> >> >> >> >> >> -----Original Message----- >> From: Tim O'Brien Sr.<[hidden email]> >> To: CONFOCALMICROSCOPY<[hidden email]> >> Sent: Tue, 29 Nov 2011 22:57 >> Subject: 3D viewing of your stacks at ASCB - Booth 853 >> >> >> ***** >> >> To join, leave or search the confocal microscopy listserv, go to: >> >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> >> ***** >> >> >> >> Dear Confocal Listserve users- >> >> >> >> If you have stacks of cells stained for tubular structures (or >> >> filaments) plus another fluorophore, and are attending ASCB, we would >> >> like to view them at the meeting in Denver. We are trying to improve >> >> our free 3D stereo rendering program (ImageSurfer 2.0) and make it a >> >> seriously useful program for Cell Biologists. >> >> >> >> With inexpensive 3D viewing systems now easily assembled (for less than >> >> $5000 including computer and monitor, glasses, etc), we feel ImageSurfer >> >> could be a great way to routinely view collocalizations within cells in >> >> many labs. The new functionality within ISurfer allows much better >> >> rendering of tubular structures (a subject dear to my heart), plus >> >> rendering of multiple colors should make it a useful tool for >> >> visualizing and quantifying relative locations of proteins within >> >> cells. The program is freely available at our website >> >> (http://cismm.cs.unc.edu/) along with many other useful programs such as >> >> the microscope simulator, image tracker, and so on. We also have a how >> >> to for assembling your own stereo viewing system for your lab. >> >> >> >> The advantage to viewing your stacks in Denver is that we can learn what >> >> you like and how we can improve it. You can view your images with 3D >> >> glasses at our booth, and take back a movie of the rendering. >> >> >> >> If you aren't going to ASCB, please feel free to download the program >> >> and try it out. Let me know what can be improved, or contact us through >> >> the website. If you are going, please consider bringing a thumb drive >> >> with your stacks to booth 853. >> >> >> >> Thanks! >> >> >> >> Tim O'Brien >> >> >> > > -- George McNamara, PhD Analytical Imaging Core Facility University of Miami |
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