20X objective for 2P work
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Boswell, Carl A - (cboswell) |
20X objective for 2P work
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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 listers, We're currently on the search for a high NA water immersion 20x objective to use for intravital 2P imaging. As such, we need something that does it all, short wavelength transmission for SHG and long wavelength transmission for our Ti:Saph excitation source. Any thoughts on what folks use/have seen/would like to have/have heard about would be appreciated. Thanks, c Carl A. Boswell, Ph.D. Molecular and Cellular Biology University of Arizona 520-954-7053 FAX 520-621-3709 |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=646 I think it may only be sold with the Flo-view1000MPE but does what you want. Patty On 10/18/2010 11:20 AM, Carl Boswell wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear listers, > We're currently on the search for a high NA water immersion 20x > objective to use for intravital 2P imaging. As such, we need > something that does it all, short wavelength transmission for SHG and > long wavelength transmission for our Ti:Saph excitation source. Any > thoughts on what folks use/have seen/would like to have/have heard > about would be appreciated. > > Thanks, > c > > Carl A. Boswell, Ph.D. > Molecular and Cellular Biology > University of Arizona > 520-954-7053 > FAX 520-621-3709 |
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Stephen Firth (Med) |
Re: 20X objective for 2P work
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** We use the Leica 20x 1.0NA dipping on our 2P (Leica CFS w. MaiTai), it's worked well for all our deep/intravital imaging. Cheers, Stephen > > On 10/18/2010 11:20 AM, Carl Boswell wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Dear listers, >> We're currently on the search for a high NA water immersion 20x objective >> to use for intravital 2P imaging. As such, we need something that does it >> all, short wavelength transmission for SHG and long wavelength transmission >> for our Ti:Saph excitation source. Any thoughts on what folks use/have >> seen/would like to have/have heard about would be appreciated. >> >> Thanks, >> c >> >> Carl A. Boswell, Ph.D. >> Molecular and Cellular Biology >> University of Arizona >> 520-954-7053 >> FAX 520-621-3709 >> > -- Stephen Firth Manager of Advanced Optical Microscopy Monash Micro Imaging Monash University Room G58 Building 75 Phone 990 55612 Mob 0400 695 425 |
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In reply to this post by PattyJansma
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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 List, Unfortunately the Olympus 25x is only sold as part of a complete system (we have one and it has been great). But the older 20x XLumPlanFL works pretty well too for visible and IR wavelengths and you should be able to buy one of those separately. Cheers Cam Cameron J. Nowell Microscopy Manager Centre for Advanced Microscopy Ludwig Institute for Cancer Research PO Box 2008 Royal Melbourne Hospital Victoria, 3050 AUSTRALIA Office: +61 3 9341 3155 Mobile: +61422882700 Fax: +61 3 9341 3104 Facility Website -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Patty Jansma Sent: Tuesday, 19 October 2010 6:07 AM To: [hidden email] Subject: Re: 20X objective for 2P work ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=6 46 I think it may only be sold with the Flo-view1000MPE but does what you want. Patty On 10/18/2010 11:20 AM, Carl Boswell wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear listers, > We're currently on the search for a high NA water immersion 20x > objective to use for intravital 2P imaging. As such, we need > something that does it all, short wavelength transmission for SHG and > long wavelength transmission for our Ti:Saph excitation source. Any > thoughts on what folks use/have seen/would like to have/have heard > about would be appreciated. > > Thanks, > c > > Carl A. Boswell, Ph.D. > Molecular and Cellular Biology > University of Arizona > 520-954-7053 > FAX 520-621-3709 This communication is intended only for the named recipient and may contain information that is confidential, legally privileged or subject to copyright; the Ludwig Institute for Cancer Research Ltd does not waive any rights if you have received this communication in error. The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. |
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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 all, Nikon also has a new 25x objective for multiphoton work - specifications look very similar to the Olympus and it available without buying a complete system... however I've not had any feedback from actual users yet. I'm hoping to be able to test one on our LaVision BioTec TriMScopes at some stage but haven't had any success getting that organised yet. We've had good results with the Olympus 20x NA 0.95 (standard and special IR versions). Regards, Adrian Smith Centenary Institute, Sydney, Australia On 19/10/2010, at 10:16 AM, Cameron Nowell wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi List, > > Unfortunately the Olympus 25x is only sold as part of a complete system > (we have one and it has been great). But the older 20x XLumPlanFL works > pretty well too for visible and IR wavelengths and you should be able to > buy one of those separately. > > > Cheers > > Cam > > > Cameron J. Nowell > Microscopy Manager > Centre for Advanced Microscopy > Ludwig Institute for Cancer Research > PO Box 2008 > Royal Melbourne Hospital > Victoria, 3050 > AUSTRALIA > Office: +61 3 9341 3155 > Mobile: +61422882700 > Fax: +61 3 9341 3104 > Facility Website > > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Patty Jansma > Sent: Tuesday, 19 October 2010 6:07 AM > To: [hidden email] > Subject: Re: 20X objective for 2P work > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=6 > 46 > > I think it may only be sold with the Flo-view1000MPE but does what you > want. > > Patty > > On 10/18/2010 11:20 AM, Carl Boswell wrote: >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Dear listers, >> We're currently on the search for a high NA water immersion 20x >> objective to use for intravital 2P imaging. As such, we need >> something that does it all, short wavelength transmission for SHG and >> long wavelength transmission for our Ti:Saph excitation source. Any >> thoughts on what folks use/have seen/would like to have/have heard >> about would be appreciated. >> >> Thanks, >> c >> >> Carl A. Boswell, Ph.D. >> Molecular and Cellular Biology >> University of Arizona >> 520-954-7053 >> FAX 520-621-3709 > > > This communication is intended only for the named recipient and may contain information that is confidential, legally privileged or subject to copyright; the Ludwig Institute for Cancer Research Ltd does not waive any rights if you have received this communication in error. > The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. |
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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, We use the 20x and its really a workhorse! I was told recently that the 20x from olympus has better response in red/infrared range than de 25x despite of the lower NA. Can anyone confirm this? All the best, Nuno Moreno, PhD Head of Equipment Management Instituto Gulbenkian de Ciência Fundação Calouste Gulbenkian Tel . +351 4464538 Fax. +351 4407970 On Oct 19, 2010, at 5:33 AM, Adrian Smith wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi all, > > Nikon also has a new 25x objective for multiphoton work - specifications look very similar to the Olympus and it available without buying a complete system... however I've not had any feedback from actual users yet. > > I'm hoping to be able to test one on our LaVision BioTec TriMScopes at some stage but haven't had any success getting that organised yet. > > We've had good results with the Olympus 20x NA 0.95 (standard and special IR versions). > > Regards, > > Adrian Smith > Centenary Institute, Sydney, Australia > > On 19/10/2010, at 10:16 AM, Cameron Nowell wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Hi List, >> >> Unfortunately the Olympus 25x is only sold as part of a complete system >> (we have one and it has been great). But the older 20x XLumPlanFL works >> pretty well too for visible and IR wavelengths and you should be able to >> buy one of those separately. >> >> >> Cheers >> >> Cam >> >> >> Cameron J. Nowell >> Microscopy Manager >> Centre for Advanced Microscopy >> Ludwig Institute for Cancer Research >> PO Box 2008 >> Royal Melbourne Hospital >> Victoria, 3050 >> AUSTRALIA >> Office: +61 3 9341 3155 >> Mobile: +61422882700 >> Fax: +61 3 9341 3104 >> Facility Website >> >> >> >> -----Original Message----- >> From: Confocal Microscopy List [mailto:[hidden email]] >> On Behalf Of Patty Jansma >> Sent: Tuesday, 19 October 2010 6:07 AM >> To: [hidden email] >> Subject: Re: 20X objective for 2P work >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=6 >> 46 >> >> I think it may only be sold with the Flo-view1000MPE but does what you >> want. >> >> Patty >> >> On 10/18/2010 11:20 AM, Carl Boswell wrote: >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> Dear listers, >>> We're currently on the search for a high NA water immersion 20x >>> objective to use for intravital 2P imaging. As such, we need >>> something that does it all, short wavelength transmission for SHG and >>> long wavelength transmission for our Ti:Saph excitation source. Any >>> thoughts on what folks use/have seen/would like to have/have heard >>> about would be appreciated. >>> >>> Thanks, >>> c >>> >>> Carl A. Boswell, Ph.D. >>> Molecular and Cellular Biology >>> University of Arizona >>> 520-954-7053 >>> FAX 520-621-3709 >> >> >> This communication is intended only for the named recipient and may contain information that is confidential, legally privileged or subject to copyright; the Ludwig Institute for Cancer Research Ltd does not waive any rights if you have received this communication in error. >> The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. |
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Andreas Bruckbauer |
Re: 20X objective for 2P work
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** >I was told recently that the 20x from olympus has better response in >red/infrared range than de 25x the 25x has a higher transmission in the red/IR. To achieve the same laser power out of the objective e.g you don't want to kill your tissue, you need to attenuate more. If you are limited in laser power, e.g. at he fringes of the spectrum and for deep imaging you get more through with the 25x. That is waht is was designed for. best wishes Andreas -----Original Message----- From: Nuno Moreno <[hidden email]> To: [hidden email] Sent: Tue, 19 Oct 2010 9:31 Subject: Re: 20X objective for 2P work ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi, We use the 20x and its really a workhorse! I was told recently that the 20x from olympus has better response in red/infrared range than de 25x despite of the lower NA. Can anyone confirm this? All the best, Nuno Moreno, PhD Head of Equipment Management Instituto Gulbenkian de Ciência Fundação Calouste Gulbenkian Tel . +351 4464538 Fax. +351 4407970 On Oct 19, 2010, at 5:33 AM, Adrian Smith wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi all, > > Nikon also has a new 25x objective for multiphoton work - specifications look very similar to the Olympus and it available without buying a complete system... however I've not had any feedback from actual users yet. > > I'm hoping to be able to test one on our LaVision BioTec TriMScopes at some stage but haven't had any success getting that organised yet. > > We've had good results with the Olympus 20x NA 0.95 (standard and special IR versions). > > Regards, > > Adrian Smith > Centenary Institute, Sydney, Australia > > On 19/10/2010, at 10:16 AM, Cameron Nowell wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Hi List, >> >> Unfortunately the Olympus 25x is only sold as part of a complete system >> (we have one and it has been great). But the older 20x XLumPlanFL works >> pretty well too for visible and IR wavelengths and you should be able to >> buy one of those separately. >> >> >> Cheers >> >> Cam >> >> >> Cameron J. Nowell >> Microscopy Manager >> Centre for Advanced Microscopy >> Ludwig Institute for Cancer Research >> PO Box 2008 >> Royal Melbourne Hospital >> Victoria, 3050 >> AUSTRALIA >> Office: +61 3 9341 3155 >> Mobile: +61422882700 >> Fax: +61 3 9341 3104 >> Facility Website >> >> >> >> -----Original Message----- >> From: Confocal Microscopy List [mailto:[hidden email]] >> On Behalf Of Patty Jansma >> Sent: Tuesday, 19 October 2010 6:07 AM >> To: [hidden email] >> Subject: Re: 20X objective for 2P work >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=6 >> 46 >> >> I think it may only be sold with the Flo-view1000MPE but does what you >> want. >> >> Patty >> >> On 10/18/2010 11:20 AM, Carl Boswell wrote: >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> Dear listers, >>> We're currently on the search for a high NA water immersion 20x >>> objective to use for intravital 2P imaging. As such, we need >>> something that does it all, short wavelength transmission for SHG and >>> long wavelength transmission for our Ti:Saph excitation source. Any >>> thoughts on what folks use/have seen/would like to have/have heard >>> about would be appreciated. >>> >>> Thanks, >>> c >>> >>> Carl A. Boswell, Ph.D. >>> Molecular and Cellular Biology >>> University of Arizona >>> 520-954-7053 >>> FAX 520-621-3709 >> >> >> This communication is intended only for the named recipient and may contain information that is confidential, legally privileged or subject to copyright; the Ludwig Institute for Cancer Research Ltd does not waive any rights if you have received this communication in error. >> The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** is there any numbers? Opinions I already have. On Oct 19, 2010, at 10:02 AM, Andreas Bruckbauer wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > > >> I was told recently that the 20x from olympus has better response in >> red/infrared range than de 25x > > the 25x has a higher transmission in the red/IR. To achieve the same laser power out of the objective e.g you don't want to kill your tissue, you need to attenuate more. If you are limited in laser power, e.g. at he fringes of the spectrum and for deep imaging you get more through with the 25x. That is waht is was designed for. > > best wishes > > Andreas > > > > > > > -----Original Message----- > From: Nuno Moreno <[hidden email]> > To: [hidden email] > Sent: Tue, 19 Oct 2010 9:31 > Subject: Re: 20X objective for 2P work > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Hi, > > > > We use the 20x and its really a workhorse! > > > > I was told recently that the 20x from olympus has better response in > > red/infrared range than de 25x despite of the lower NA. Can anyone confirm this? > > > > All the best, > > Nuno Moreno, PhD > > Head of Equipment Management > > Instituto Gulbenkian de Ciência > > Fundação Calouste Gulbenkian > > Tel . +351 4464538 > > Fax. +351 4407970 > > > > > > > > > > > > On Oct 19, 2010, at 5:33 AM, Adrian Smith wrote: > > > >> ***** > >> To join, leave or search the confocal microscopy listserv, go to: > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> ***** > >> > >> Hi all, > >> > >> Nikon also has a new 25x objective for multiphoton work - specifications look > > very similar to the Olympus and it available without buying a complete > > system... however I've not had any feedback from actual users yet. > >> > >> I'm hoping to be able to test one on our LaVision BioTec TriMScopes at some > > stage but haven't had any success getting that organised yet. > >> > >> We've had good results with the Olympus 20x NA 0.95 (standard and special IR > > versions). > >> > >> Regards, > >> > >> Adrian Smith > >> Centenary Institute, Sydney, Australia > >> > >> On 19/10/2010, at 10:16 AM, Cameron Nowell wrote: > >> > >>> ***** > >>> To join, leave or search the confocal microscopy listserv, go to: > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >>> ***** > >>> > >>> Hi List, > >>> > >>> Unfortunately the Olympus 25x is only sold as part of a complete system > >>> (we have one and it has been great). But the older 20x XLumPlanFL works > >>> pretty well too for visible and IR wavelengths and you should be able to > >>> buy one of those separately. > >>> > >>> > >>> Cheers > >>> > >>> Cam > >>> > >>> > >>> Cameron J. Nowell > >>> Microscopy Manager > >>> Centre for Advanced Microscopy > >>> Ludwig Institute for Cancer Research > >>> PO Box 2008 > >>> Royal Melbourne Hospital > >>> Victoria, 3050 > >>> AUSTRALIA > >>> Office: +61 3 9341 3155 > >>> Mobile: +61422882700 > >>> Fax: +61 3 9341 3104 > >>> Facility Website > >>> > >>> > >>> > >>> -----Original Message----- > >>> From: Confocal Microscopy List [mailto:[hidden email]] > >>> On Behalf Of Patty Jansma > >>> Sent: Tuesday, 19 October 2010 6:07 AM > >>> To: [hidden email] > >>> Subject: Re: 20X objective for 2P work > >>> > >>> ***** > >>> To join, leave or search the confocal microscopy listserv, go to: > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >>> ***** > >>> > >>> http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=6 > >>> 46 > >>> > >>> I think it may only be sold with the Flo-view1000MPE but does what you > >>> want. > >>> > >>> Patty > >>> > >>> On 10/18/2010 11:20 AM, Carl Boswell wrote: > >>>> ***** > >>>> To join, leave or search the confocal microscopy listserv, go to: > >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >>>> ***** > >>>> > >>>> Dear listers, > >>>> We're currently on the search for a high NA water immersion 20x > >>>> objective to use for intravital 2P imaging. As such, we need > >>>> something that does it all, short wavelength transmission for SHG and > >>>> long wavelength transmission for our Ti:Saph excitation source. Any > >>>> thoughts on what folks use/have seen/would like to have/have heard > >>>> about would be appreciated. > >>>> > >>>> Thanks, > >>>> c > >>>> > >>>> Carl A. Boswell, Ph.D. > >>>> Molecular and Cellular Biology > >>>> University of Arizona > >>>> 520-954-7053 > >>>> FAX 520-621-3709 > >>> > >>> > >>> This communication is intended only for the named recipient and may contain > > information that is confidential, legally privileged or subject to copyright; > > the Ludwig Institute for Cancer Research Ltd does not waive any rights if you > > have received this communication in error. > >>> The views expressed in this communication are those of the sender and do not > > necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. > > > |
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Andreas Bruckbauer |
Re: 20X objective for 2P work
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** >is there any numbers? Opinions I already have. at 800 nm, 2.5% laser power setting (AOM transmission) 25x: 10.4 mW, 20x: 7.9 mW at 950 nm: 25x: 3.2 mW, 20x: 2.85 at 1020 mW 6% 25x: 2.61, 20x 2.39 mW these were measured on the Olympus system which fills the back aperture of the 25x objective, but underfills the 20x objective, when i set the beam expander to get the same fill factor for the 25x as for the 20x, i measure 11.8 mW (800nm), 3.5mW (950), 2.91 mW (1020) so i conclude about 20-50% more transmission in the IR for the 25x These values were measured in air, so there is some total reflection expected for the 25x NA1.05 objective. The measured power levels also depend on the output characteristics of the laser. Furthermore there are some uncertainties depending on the type of power meter used, i think this was done with a photodiode based sensor (was a while ago). best wishes Andreas On Oct 19, 2010, at 10:02 AM, Andreas Bruckbauer wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > > >> I was told recently that the 20x from olympus has better response in >> red/infrared range than de 25x > > the 25x has a higher transmission in the red/IR. To achieve the same laser power out of the objective e.g you don't want to kill your tissue, you need to attenuate more. If you are limited in laser power, e.g. at he fringes of the spectrum and for deep imaging you get more through with the 25x. That is waht is was designed for. > > best wishes > > Andreas > > > > > > > -----Original Message----- > From: Nuno Moreno <[hidden email]> > To: [hidden email] > Sent: Tue, 19 Oct 2010 9:31 > Subject: Re: 20X objective for 2P work > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > Hi, > > > > We use the 20x and its really a workhorse! > > > > I was told recently that the 20x from olympus has better response in > > red/infrared range than de 25x despite of the lower NA. Can anyone confirm this? > > > > All the best, > > Nuno Moreno, PhD > > Head of Equipment Management > > Instituto Gulbenkian de Ciência > > Fundação Calouste Gulbenkian > > Tel . +351 4464538 > > Fax. +351 4407970 > > > > > > > > > > > > On Oct 19, 2010, at 5:33 AM, Adrian Smith wrote: > > > >> ***** > >> To join, leave or search the confocal microscopy listserv, go to: > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> ***** > >> > >> Hi all, > >> > >> Nikon also has a new 25x objective for multiphoton work - specifications look > > very similar to the Olympus and it available without buying a complete > > system... however I've not had any feedback from actual users yet. > >> > >> I'm hoping to be able to test one on our LaVision BioTec TriMScopes at some > > stage but haven't had any success getting that organised yet. > >> > >> We've had good results with the Olympus 20x NA 0.95 (standard and special IR > > versions). > >> > >> Regards, > >> > >> Adrian Smith > >> Centenary Institute, Sydney, Australia > >> > >> On 19/10/2010, at 10:16 AM, Cameron Nowell wrote: > >> > >>> ***** > >>> To join, leave or search the confocal microscopy listserv, go to: > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >>> ***** > >>> > >>> Hi List, > >>> > >>> Unfortunately the Olympus 25x is only sold as part of a complete system > >>> (we have one and it has been great). But the older 20x XLumPlanFL works > >>> pretty well too for visible and IR wavelengths and you should be able to > >>> buy one of those separately. > >>> > >>> > >>> Cheers > >>> > >>> Cam > >>> > >>> > >>> Cameron J. Nowell > >>> Microscopy Manager > >>> Centre for Advanced Microscopy > >>> Ludwig Institute for Cancer Research > >>> PO Box 2008 > >>> Royal Melbourne Hospital > >>> Victoria, 3050 > >>> AUSTRALIA > >>> Office: +61 3 9341 3155 > >>> Mobile: +61422882700 > >>> Fax: +61 3 9341 3104 > >>> Facility Website > >>> > >>> > >>> > >>> -----Original Message----- > >>> From: Confocal Microscopy List [mailto:[hidden email]] > >>> On Behalf Of Patty Jansma > >>> Sent: Tuesday, 19 October 2010 6:07 AM > >>> To: [hidden email] > >>> Subject: Re: 20X objective for 2P work > >>> > >>> ***** > >>> To join, leave or search the confocal microscopy listserv, go to: > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >>> ***** > >>> > >>> http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=6 > >>> 46 > >>> > >>> I think it may only be sold with the Flo-view1000MPE but does what you > >>> want. > >>> > >>> Patty > >>> > >>> On 10/18/2010 11:20 AM, Carl Boswell wrote: > >>>> ***** > >>>> To join, leave or search the confocal microscopy listserv, go to: > >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >>>> ***** > >>>> > >>>> Dear listers, > >>>> We're currently on the search for a high NA water immersion 20x > >>>> objective to use for intravital 2P imaging. As such, we need > >>>> something that does it all, short wavelength transmission for SHG and > >>>> long wavelength transmission for our Ti:Saph excitation source. Any > >>>> thoughts on what folks use/have seen/would like to have/have heard > >>>> about would be appreciated. > >>>> > >>>> Thanks, > >>>> c > >>>> > >>>> Carl A. Boswell, Ph.D. > >>>> Molecular and Cellular Biology > >>>> University of Arizona > >>>> 520-954-7053 > >>>> FAX 520-621-3709 > >>> > >>> > >>> This communication is intended only for the named recipient and may contain > > information that is confidential, legally privileged or subject to copyright; > > the Ludwig Institute for Cancer Research Ltd does not waive any rights if you > > have received this communication in error. > >>> The views expressed in this communication are those of the sender and do not > > necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. > > > |
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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, Now I think I understand why on Nimejen they have the 20x. They have an OPO and use >1100 nm for mcherry. It might be that there is a turning point in performance because at 1020 the difference is only 9%. Thank you for those numbers. Even with technical limitations better those than nothing! NM On Oct 19, 2010, at 4:28 PM, Andreas Bruckbauer wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > >> is there any numbers? Opinions I already have. > > > > at 800 nm, 2.5% laser power setting (AOM transmission) > 25x: 10.4 mW, 20x: 7.9 mW > > at 950 nm: > 25x: 3.2 mW, 20x: 2.85 > > at 1020 mW 6% > 25x: 2.61, 20x 2.39 mW > > these were measured on the Olympus system which fills the back aperture > of the 25x objective, but underfills the 20x objective, when i set the beam expander to get the same fill factor for the 25x as for the 20x, i measure > 11.8 mW (800nm), 3.5mW (950), 2.91 mW (1020) > > so i conclude about 20-50% more transmission in the IR for the 25x > These values were measured in air, so there is some total reflection expected for the 25x NA1.05 objective. The measured power levels also depend on > the output characteristics of the laser. Furthermore there are some uncertainties depending on the type of power meter used, i think this was done with > a photodiode based sensor (was a while ago). > > best wishes > > Andreas > > > > > > > > On Oct 19, 2010, at 10:02 AM, Andreas Bruckbauer wrote: > > > >> ***** > >> To join, leave or search the confocal microscopy listserv, go to: > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> ***** > >> > >> > >> > >>> I was told recently that the 20x from olympus has better response in > >>> red/infrared range than de 25x > >> > >> the 25x has a higher transmission in the red/IR. To achieve the same laser > > power out of the objective e.g you don't want to kill your tissue, you need to > > attenuate more. If you are limited in laser power, e.g. at he fringes of the > > spectrum and for deep imaging you get more through with the 25x. That is waht is > > was designed for. > >> > >> best wishes > >> > >> Andreas > >> > >> > >> > >> > >> > >> > >> -----Original Message----- > >> From: Nuno Moreno <[hidden email]> > >> To: [hidden email] > >> Sent: Tue, 19 Oct 2010 9:31 > >> Subject: Re: 20X objective for 2P work > >> > >> > >> ***** > >> > >> To join, leave or search the confocal microscopy listserv, go to: > >> > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >> ***** > >> > >> > >> > >> Hi, > >> > >> > >> > >> We use the 20x and its really a workhorse! > >> > >> > >> > >> I was told recently that the 20x from olympus has better response in > >> > >> red/infrared range than de 25x despite of the lower NA. Can anyone confirm > > this? > >> > >> > >> > >> All the best, > >> > >> Nuno Moreno, PhD > >> > >> Head of Equipment Management > >> > >> Instituto Gulbenkian de Ciência > >> > >> Fundação Calouste Gulbenkian > >> > >> Tel . +351 4464538 > >> > >> Fax. +351 4407970 > >> > >> > >> > >> > >> > >> > >> > >> > >> > >> > >> > >> On Oct 19, 2010, at 5:33 AM, Adrian Smith wrote: > >> > >> > >> > >>> ***** > >> > >>> To join, leave or search the confocal microscopy listserv, go to: > >> > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >>> ***** > >> > >>> > >> > >>> Hi all, > >> > >>> > >> > >>> Nikon also has a new 25x objective for multiphoton work - specifications look > > > >> > >> very similar to the Olympus and it available without buying a complete > >> > >> system... however I've not had any feedback from actual users yet. > >> > >>> > >> > >>> I'm hoping to be able to test one on our LaVision BioTec TriMScopes at some > >> > >> stage but haven't had any success getting that organised yet. > >> > >>> > >> > >>> We've had good results with the Olympus 20x NA 0.95 (standard and special IR > >> > >> versions). > >> > >>> > >> > >>> Regards, > >> > >>> > >> > >>> Adrian Smith > >> > >>> Centenary Institute, Sydney, Australia > >> > >>> > >> > >>> On 19/10/2010, at 10:16 AM, Cameron Nowell wrote: > >> > >>> > >> > >>>> ***** > >> > >>>> To join, leave or search the confocal microscopy listserv, go to: > >> > >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >>>> ***** > >> > >>>> > >> > >>>> Hi List, > >> > >>>> > >> > >>>> Unfortunately the Olympus 25x is only sold as part of a complete system > >> > >>>> (we have one and it has been great). But the older 20x XLumPlanFL works > >> > >>>> pretty well too for visible and IR wavelengths and you should be able to > >> > >>>> buy one of those separately. > >> > >>>> > >> > >>>> > >> > >>>> Cheers > >> > >>>> > >> > >>>> Cam > >> > >>>> > >> > >>>> > >> > >>>> Cameron J. Nowell > >> > >>>> Microscopy Manager > >> > >>>> Centre for Advanced Microscopy > >> > >>>> Ludwig Institute for Cancer Research > >> > >>>> PO Box 2008 > >> > >>>> Royal Melbourne Hospital > >> > >>>> Victoria, 3050 > >> > >>>> AUSTRALIA > >> > >>>> Office: +61 3 9341 3155 > >> > >>>> Mobile: +61422882700 > >> > >>>> Fax: +61 3 9341 3104 > >> > >>>> Facility Website > >> > >>>> > >> > >>>> > >> > >>>> > >> > >>>> -----Original Message----- > >> > >>>> From: Confocal Microscopy List [mailto:[hidden email]] > >> > >>>> On Behalf Of Patty Jansma > >> > >>>> Sent: Tuesday, 19 October 2010 6:07 AM > >> > >>>> To: [hidden email] > >> > >>>> Subject: Re: 20X objective for 2P work > >> > >>>> > >> > >>>> ***** > >> > >>>> To join, leave or search the confocal microscopy listserv, go to: > >> > >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >>>> ***** > >> > >>>> > >> > >>>> http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=6 > >> > >>>> 46 > >> > >>>> > >> > >>>> I think it may only be sold with the Flo-view1000MPE but does what you > >> > >>>> want. > >> > >>>> > >> > >>>> Patty > >> > >>>> > >> > >>>> On 10/18/2010 11:20 AM, Carl Boswell wrote: > >> > >>>>> ***** > >> > >>>>> To join, leave or search the confocal microscopy listserv, go to: > >> > >>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >>>>> ***** > >> > >>>>> > >> > >>>>> Dear listers, > >> > >>>>> We're currently on the search for a high NA water immersion 20x > >> > >>>>> objective to use for intravital 2P imaging. As such, we need > >> > >>>>> something that does it all, short wavelength transmission for SHG and > >> > >>>>> long wavelength transmission for our Ti:Saph excitation source. Any > >> > >>>>> thoughts on what folks use/have seen/would like to have/have heard > >> > >>>>> about would be appreciated. > >> > >>>>> > >> > >>>>> Thanks, > >> > >>>>> c > >> > >>>>> > >> > >>>>> Carl A. Boswell, Ph.D. > >> > >>>>> Molecular and Cellular Biology > >> > >>>>> University of Arizona > >> > >>>>> 520-954-7053 > >> > >>>>> FAX 520-621-3709 > >> > >>>> > >> > >>>> > >> > >>>> This communication is intended only for the named recipient and may contain > >> > >> information that is confidential, legally privileged or subject to copyright; > >> > >> the Ludwig Institute for Cancer Research Ltd does not waive any rights if you > >> > >> have received this communication in error. > >> > >>>> The views expressed in this communication are those of the sender and do not > > > >> > >> necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. > >> > >> > >> > > > |
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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, If you are referring to the LaVision BioTec systems at Nijmegen then the other reason they will have the 20x is that Olympus won't sell them the 25x as it is not an Olympus system :) We are in the same boat here with our LaVision BioTec systems with OPO. We actually have two versions of the 20x Olympus objective - one "normal" and one "special IR" which has much better transmission in the longer wavelengths. I don't have any rigorous numbers or 25x to compare with so I won't post anything here. We have also heard that the current 20x Olympus objectives (might be NA 1.0?) are not as good for the OPO as the older NA0.95 special IR version... so it certainly seems that not all Olympus 20x objectives are created equal! Regards, Adrian Smith Centenary Institute, Sydney, Australia On 20/10/2010, at 3:05 AM, Nuno Moreno wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi, > > Now I think I understand why on Nimejen they have the 20x. They have an OPO and use >1100 nm for mcherry. It might be that there is a turning point in performance because at 1020 the difference is only 9%. > > Thank you for those numbers. Even with technical limitations better those than nothing! > NM > > > > > On Oct 19, 2010, at 4:28 PM, Andreas Bruckbauer wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >>> is there any numbers? Opinions I already have. >> >> >> >> at 800 nm, 2.5% laser power setting (AOM transmission) >> 25x: 10.4 mW, 20x: 7.9 mW >> >> at 950 nm: >> 25x: 3.2 mW, 20x: 2.85 >> >> at 1020 mW 6% >> 25x: 2.61, 20x 2.39 mW >> >> these were measured on the Olympus system which fills the back aperture >> of the 25x objective, but underfills the 20x objective, when i set the beam expander to get the same fill factor for the 25x as for the 20x, i measure >> 11.8 mW (800nm), 3.5mW (950), 2.91 mW (1020) >> >> so i conclude about 20-50% more transmission in the IR for the 25x >> These values were measured in air, so there is some total reflection expected for the 25x NA1.05 objective. The measured power levels also depend on >> the output characteristics of the laser. Furthermore there are some uncertainties depending on the type of power meter used, i think this was done with >> a photodiode based sensor (was a while ago). >> >> best wishes >> >> Andreas >> >> >> >> >> >> >> >> On Oct 19, 2010, at 10:02 AM, Andreas Bruckbauer wrote: >> >> >> >>> ***** >> >>> To join, leave or search the confocal microscopy listserv, go to: >> >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> >>> ***** >> >>> >> >>> >> >>> >> >>>> I was told recently that the 20x from olympus has better response in >> >>>> red/infrared range than de 25x >> >>> >> >>> the 25x has a higher transmission in the red/IR. To achieve the same laser >> >> power out of the objective e.g you don't want to kill your tissue, you need to >> >> attenuate more. If you are limited in laser power, e.g. at he fringes of the >> >> spectrum and for deep imaging you get more through with the 25x. That is waht is >> >> was designed for. >> >>> >> >>> best wishes >> >>> >> >>> Andreas >> >>> >> >>> >> >>> >> >>> >> >>> >> >>> >> >>> -----Original Message----- >> >>> From: Nuno Moreno <[hidden email]> >> >>> To: [hidden email] >> >>> Sent: Tue, 19 Oct 2010 9:31 >> >>> Subject: Re: 20X objective for 2P work >> >>> >> >>> >> >>> ***** >> >>> >> >>> To join, leave or search the confocal microscopy listserv, go to: >> >>> >> >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> >>> >> >>> ***** >> >>> >> >>> >> >>> >> >>> Hi, >> >>> >> >>> >> >>> >> >>> We use the 20x and its really a workhorse! >> >>> >> >>> >> >>> >> >>> I was told recently that the 20x from olympus has better response in >> >>> >> >>> red/infrared range than de 25x despite of the lower NA. Can anyone confirm >> >> this? >> >>> >> >>> >> >>> >> >>> All the best, >> >>> >> >>> Nuno Moreno, PhD >> >>> >> >>> Head of Equipment Management >> >>> >> >>> Instituto Gulbenkian de Ciência >> >>> >> >>> Fundação Calouste Gulbenkian >> >>> >> >>> Tel . +351 4464538 >> >>> >> >>> Fax. +351 4407970 >> >>> >> >>> >> >>> >> >>> >> >>> >> >>> >> >>> >> >>> >> >>> >> >>> >> >>> >> >>> On Oct 19, 2010, at 5:33 AM, Adrian Smith wrote: >> >>> >> >>> >> >>> >> >>>> ***** >> >>> >> >>>> To join, leave or search the confocal microscopy listserv, go to: >> >>> >> >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> >>> >> >>>> ***** >> >>> >> >>>> >> >>> >> >>>> Hi all, >> >>> >> >>>> >> >>> >> >>>> Nikon also has a new 25x objective for multiphoton work - specifications look >> >> >> >>> >> >>> very similar to the Olympus and it available without buying a complete >> >>> >> >>> system... however I've not had any feedback from actual users yet. >> >>> >> >>>> >> >>> >> >>>> I'm hoping to be able to test one on our LaVision BioTec TriMScopes at some >> >>> >> >>> stage but haven't had any success getting that organised yet. >> >>> >> >>>> >> >>> >> >>>> We've had good results with the Olympus 20x NA 0.95 (standard and special IR >> >>> >> >>> versions). >> >>> >> >>>> >> >>> >> >>>> Regards, >> >>> >> >>>> >> >>> >> >>>> Adrian Smith >> >>> >> >>>> Centenary Institute, Sydney, Australia >> >>> >> >>>> >> >>> >> >>>> On 19/10/2010, at 10:16 AM, Cameron Nowell wrote: >> >>> >> >>>> >> >>> >> >>>>> ***** >> >>> >> >>>>> To join, leave or search the confocal microscopy listserv, go to: >> >>> >> >>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> >>> >> >>>>> ***** >> >>> >> >>>>> >> >>> >> >>>>> Hi List, >> >>> >> >>>>> >> >>> >> >>>>> Unfortunately the Olympus 25x is only sold as part of a complete system >> >>> >> >>>>> (we have one and it has been great). But the older 20x XLumPlanFL works >> >>> >> >>>>> pretty well too for visible and IR wavelengths and you should be able to >> >>> >> >>>>> buy one of those separately. >> >>> >> >>>>> >> >>> >> >>>>> >> >>> >> >>>>> Cheers >> >>> >> >>>>> >> >>> >> >>>>> Cam >> >>> >> >>>>> >> >>> >> >>>>> >> >>> >> >>>>> Cameron J. Nowell >> >>> >> >>>>> Microscopy Manager >> >>> >> >>>>> Centre for Advanced Microscopy >> >>> >> >>>>> Ludwig Institute for Cancer Research >> >>> >> >>>>> PO Box 2008 >> >>> >> >>>>> Royal Melbourne Hospital >> >>> >> >>>>> Victoria, 3050 >> >>> >> >>>>> AUSTRALIA >> >>> >> >>>>> Office: +61 3 9341 3155 >> >>> >> >>>>> Mobile: +61422882700 >> >>> >> >>>>> Fax: +61 3 9341 3104 >> >>> >> >>>>> Facility Website >> >>> >> >>>>> >> >>> >> >>>>> >> >>> >> >>>>> >> >>> >> >>>>> -----Original Message----- >> >>> >> >>>>> From: Confocal Microscopy List [mailto:[hidden email]] >> >>> >> >>>>> On Behalf Of Patty Jansma >> >>> >> >>>>> Sent: Tuesday, 19 October 2010 6:07 AM >> >>> >> >>>>> To: [hidden email] >> >>> >> >>>>> Subject: Re: 20X objective for 2P work >> >>> >> >>>>> >> >>> >> >>>>> ***** >> >>> >> >>>>> To join, leave or search the confocal microscopy listserv, go to: >> >>> >> >>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> >>> >> >>>>> ***** >> >>> >> >>>>> >> >>> >> >>>>> http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=6 >> >>> >> >>>>> 46 >> >>> >> >>>>> >> >>> >> >>>>> I think it may only be sold with the Flo-view1000MPE but does what you >> >>> >> >>>>> want. >> >>> >> >>>>> >> >>> >> >>>>> Patty >> >>> >> >>>>> >> >>> >> >>>>> On 10/18/2010 11:20 AM, Carl Boswell wrote: >> >>> >> >>>>>> ***** >> >>> >> >>>>>> To join, leave or search the confocal microscopy listserv, go to: >> >>> >> >>>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> >>> >> >>>>>> ***** >> >>> >> >>>>>> >> >>> >> >>>>>> Dear listers, >> >>> >> >>>>>> We're currently on the search for a high NA water immersion 20x >> >>> >> >>>>>> objective to use for intravital 2P imaging. As such, we need >> >>> >> >>>>>> something that does it all, short wavelength transmission for SHG and >> >>> >> >>>>>> long wavelength transmission for our Ti:Saph excitation source. Any >> >>> >> >>>>>> thoughts on what folks use/have seen/would like to have/have heard >> >>> >> >>>>>> about would be appreciated. >> >>> >> >>>>>> >> >>> >> >>>>>> Thanks, >> >>> >> >>>>>> c >> >>> >> >>>>>> >> >>> >> >>>>>> Carl A. Boswell, Ph.D. >> >>> >> >>>>>> Molecular and Cellular Biology >> >>> >> >>>>>> University of Arizona >> >>> >> >>>>>> 520-954-7053 >> >>> >> >>>>>> FAX 520-621-3709 >> >>> >> >>>>> >> >>> >> >>>>> >> >>> >> >>>>> This communication is intended only for the named recipient and may contain >> >>> >> >>> information that is confidential, legally privileged or subject to copyright; >> >>> >> >>> the Ludwig Institute for Cancer Research Ltd does not waive any rights if you >> >>> >> >>> have received this communication in error. >> >>> >> >>>>> The views expressed in this communication are those of the sender and do not >> >> >> >>> >> >>> necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. >> >>> >> >>> >> >>> >> >> >> |
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Andreas Bruckbauer |
Re: 20X objective for 2P work
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In reply to this post by Nuno Moreno
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** OLympus does not have the transmission data for the 25x NA 1.05 objective (its called XLPLN25xW-MP) online. When i remember correctly from prensentation by Eiji Yokoi at the FOM 2008 where they launched this lens, the transmission was improved over the whole range of wavelengths. My measurement was with the old 20x, XLUMPLFL 20XW (NA0.95) and now this is redesigned and called XLUMPLFLN 20XW (NA 1.0), the transmission curve up to 1000 nm is available on the Olympus website http://microscope.olympus-global.com/uis2/en/xlumplfln20xw/ Maybe Olympus can comment on the transmission > 1100 nm for all these objectives? Another issue when using these lenses on other manufacturers microscopes and these include Olympus microscope stands sold to other manufacturers such as Lavision, that they might have different scanning lenses or tube lenses leading to different results. best wishes Andreas -----Original Message----- From: Nuno Moreno <[hidden email]> To: [hidden email] Sent: Tue, 19 Oct 2010 17:05 Subject: Re: 20X objective for 2P work ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi, Now I think I understand why on Nimejen they have the 20x. They have an OPO and use >1100 nm for mcherry. It might be that there is a turning point in performance because at 1020 the difference is only 9%. Thank you for those numbers. Even with technical limitations better those than nothing! NM On Oct 19, 2010, at 4:28 PM, Andreas Bruckbauer wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > >> is there any numbers? Opinions I already have. > > > > at 800 nm, 2.5% laser power setting (AOM transmission) > 25x: 10.4 mW, 20x: 7.9 mW > > at 950 nm: > 25x: 3.2 mW, 20x: 2.85 > > at 1020 mW 6% > 25x: 2.61, 20x 2.39 mW > > these were measured on the Olympus system which fills the back aperture > of the 25x objective, but underfills the 20x objective, when i set the beam expander to get the same fill factor for the 25x as for the 20x, i measure > 11.8 mW (800nm), 3.5mW (950), 2.91 mW (1020) > > so i conclude about 20-50% more transmission in the IR for the 25x > These values were measured in air, so there is some total reflection expected for the 25x NA1.05 objective. The measured power levels also depend on > the output characteristics of the laser. Furthermore there are some uncertainties depending on the type of power meter used, i think this was done with > a photodiode based sensor (was a while ago). > > best wishes > > Andreas > > > > > > > > On Oct 19, 2010, at 10:02 AM, Andreas Bruckbauer wrote: > > > >> ***** > >> To join, leave or search the confocal microscopy listserv, go to: > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> ***** > >> > >> > >> > >>> I was told recently that the 20x from olympus has better response in > >>> red/infrared range than de 25x > >> > >> the 25x has a higher transmission in the red/IR. To achieve the same laser > > power out of the objective e.g you don't want to kill your tissue, you need to > > attenuate more. If you are limited in laser power, e.g. at he fringes of the > > spectrum and for deep imaging you get more through with the 25x. That is waht is > > was designed for. > >> > >> best wishes > >> > >> Andreas > >> > >> > >> > >> > >> > >> > >> -----Original Message----- > >> From: Nuno Moreno <[hidden email]> > >> To: [hidden email] > >> Sent: Tue, 19 Oct 2010 9:31 > >> Subject: Re: 20X objective for 2P work > >> > >> > >> ***** > >> > >> To join, leave or search the confocal microscopy listserv, go to: > >> > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >> ***** > >> > >> > >> > >> Hi, > >> > >> > >> > >> We use the 20x and its really a workhorse! > >> > >> > >> > >> I was told recently that the 20x from olympus has better response in > >> > >> red/infrared range than de 25x despite of the lower NA. Can anyone confirm > > this? > >> > >> > >> > >> All the best, > >> > >> Nuno Moreno, PhD > >> > >> Head of Equipment Management > >> > >> Instituto Gulbenkian de Ciência > >> > >> Fundação Calouste Gulbenkian > >> > >> Tel . +351 4464538 > >> > >> Fax. +351 4407970 > >> > >> > >> > >> > >> > >> > >> > >> > >> > >> > >> > >> On Oct 19, 2010, at 5:33 AM, Adrian Smith wrote: > >> > >> > >> > >>> ***** > >> > >>> To join, leave or search the confocal microscopy listserv, go to: > >> > >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >>> ***** > >> > >>> > >> > >>> Hi all, > >> > >>> > >> > >>> Nikon also has a new 25x objective for multiphoton work - specifications look > > > >> > >> very similar to the Olympus and it available without buying a complete > >> > >> system... however I've not had any feedback from actual users yet. > >> > >>> > >> > >>> I'm hoping to be able to test one on our LaVision BioTec TriMScopes at some > >> > >> stage but haven't had any success getting that organised yet. > >> > >>> > >> > >>> We've had good results with the Olympus 20x NA 0.95 (standard and special IR > >> > >> versions). > >> > >>> > >> > >>> Regards, > >> > >>> > >> > >>> Adrian Smith > >> > >>> Centenary Institute, Sydney, Australia > >> > >>> > >> > >>> On 19/10/2010, at 10:16 AM, Cameron Nowell wrote: > >> > >>> > >> > >>>> ***** > >> > >>>> To join, leave or search the confocal microscopy listserv, go to: > >> > >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >>>> ***** > >> > >>>> > >> > >>>> Hi List, > >> > >>>> > >> > >>>> Unfortunately the Olympus 25x is only sold as part of a complete system > >> > >>>> (we have one and it has been great). But the older 20x XLumPlanFL works > >> > >>>> pretty well too for visible and IR wavelengths and you should be able to > >> > >>>> buy one of those separately. > >> > >>>> > >> > >>>> > >> > >>>> Cheers > >> > >>>> > >> > >>>> Cam > >> > >>>> > >> > >>>> > >> > >>>> Cameron J. Nowell > >> > >>>> Microscopy Manager > >> > >>>> Centre for Advanced Microscopy > >> > >>>> Ludwig Institute for Cancer Research > >> > >>>> PO Box 2008 > >> > >>>> Royal Melbourne Hospital > >> > >>>> Victoria, 3050 > >> > >>>> AUSTRALIA > >> > >>>> Office: +61 3 9341 3155 > >> > >>>> Mobile: +61422882700 > >> > >>>> Fax: +61 3 9341 3104 > >> > >>>> Facility Website > >> > >>>> > >> > >>>> > >> > >>>> > >> > >>>> -----Original Message----- > >> > >>>> From: Confocal Microscopy List [mailto:[hidden email]] > >> > >>>> On Behalf Of Patty Jansma > >> > >>>> Sent: Tuesday, 19 October 2010 6:07 AM > >> > >>>> To: [hidden email] > >> > >>>> Subject: Re: 20X objective for 2P work > >> > >>>> > >> > >>>> ***** > >> > >>>> To join, leave or search the confocal microscopy listserv, go to: > >> > >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >>>> ***** > >> > >>>> > >> > >>>> http://www.olympusamerica.com/seg_section/seg_pressDetails.asp?pressNo=6 > >> > >>>> 46 > >> > >>>> > >> > >>>> I think it may only be sold with the Flo-view1000MPE but does what you > >> > >>>> want. > >> > >>>> > >> > >>>> Patty > >> > >>>> > >> > >>>> On 10/18/2010 11:20 AM, Carl Boswell wrote: > >> > >>>>> ***** > >> > >>>>> To join, leave or search the confocal microscopy listserv, go to: > >> > >>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> > >>>>> ***** > >> > >>>>> > >> > >>>>> Dear listers, > >> > >>>>> We're currently on the search for a high NA water immersion 20x > >> > >>>>> objective to use for intravital 2P imaging. As such, we need > >> > >>>>> something that does it all, short wavelength transmission for SHG and > >> > >>>>> long wavelength transmission for our Ti:Saph excitation source. Any > >> > >>>>> thoughts on what folks use/have seen/would like to have/have heard > >> > >>>>> about would be appreciated. > >> > >>>>> > >> > >>>>> Thanks, > >> > >>>>> c > >> > >>>>> > >> > >>>>> Carl A. Boswell, Ph.D. > >> > >>>>> Molecular and Cellular Biology > >> > >>>>> University of Arizona > >> > >>>>> 520-954-7053 > >> > >>>>> FAX 520-621-3709 > >> > >>>> > >> > >>>> > >> > >>>> This communication is intended only for the named recipient and may contain > >> > >> information that is confidential, legally privileged or subject to copyright; > >> > >> the Ludwig Institute for Cancer Research Ltd does not waive any rights if you > >> > >> have received this communication in error. > >> > >>>> The views expressed in this communication are those of the sender and do not > > > >> > >> necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd. > >> > >> > >> > > > |
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Steffen Dietzel |
Re: 20X objective for 2P work
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In reply to this post by Adrian Smith-6
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Adrian is right, there was a change in manufacturing for the Olympus 20x, I believe in 2009. Before, there was the XLUMPLFL20x NA 0.95 and the specially coated IR Version IR-XLUMPLFL20x, which is still used by us and others for multi-photon microscopy (although originally designed for IR-DIC). Instead of those then discontinued lenses, the XLUMPLFLN20x (note the N) was then offered. I once got the following transmission numbers from an Olympus representative, didn't verify them by measuring myself: 500 nm 650nm 800nm 1200 nm XLUMPLFL20x 0.73 0.66 0.60 0.50 XLUMPLFL20x-IR 0.42 0.61 0.74 0.69 XLUMPLFLN20x 0.77 0.77 0.68 0.52 So, for a two-photon process with backward-detected 500 nm emission, if you do the math the new one should be better than both old versions, essentially because you loose less of the emitted photons: (exc^2 * em.) 800/500 1200/500 1200/650 XLUMPLFL20x-IR 0.23 0,2 0.29 XLUMPLFLN20x 0.35 0,21 0.21 This predicted outcome for 500 nm emission was used by Olympus to argue that an IR-coated version of the new lens is not needed. However, if you are interessted in three-photon processes and/or forward detection (eg. THG), then you should keep a close eye on any still available old IR 20x. Same if you go for two-photon fluorescence with red dyes. As far as I know, there is no IR-coated version of the new objective (yet), hopefully I am wrong. Maybe if we push this a little. While we are on the objectives: Did any one of you bothered to measure the chromatic aberration for Ti:Sa lasers and OPOs? For our system (LaVision BioTech) with the XLUMPLFL20x-IR, I measured a z-shift of the focal plane of about 9 µm between 725 and 1050 and about 7 µm between 1175 and 1300 nm. I am wondering if this is in the normal range. Steffen On 20.10.2010 01:11, Adrian Smith wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Hi, > > If you are referring to the LaVision BioTec systems at Nijmegen then the other reason they will have the 20x is that Olympus won't sell them the 25x as it is not an Olympus system :) > > We are in the same boat here with our LaVision BioTec systems with OPO. > > We actually have two versions of the 20x Olympus objective - one "normal" and one "special IR" which has much better transmission in the longer wavelengths. I don't have any rigorous numbers or 25x to compare with so I won't post anything here. > > We have also heard that the current 20x Olympus objectives (might be NA 1.0?) are not as good for the OPO as the older NA0.95 special IR version... so it certainly seems that not all Olympus 20x objectives are created equal! > > Regards, > > Adrian Smith > Centenary Institute, Sydney, Australia > > ------------------------------------------------------------ Steffen Dietzel, PD Dr. rer. nat Ludwig-Maximilians-Universität München Walter-Brendel-Zentrum für experimentelle Medizin (WBex) Head of light microscopy |
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