light sheet feedback
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear list We are currently preparing to tender for a light sheet system for our imaging facility. The aim is to image 2 types of samples: - Tissue chunks, cleared with Clarity (mostly Fructose) and fluorescent, max @ 3x3x3 mm - Live organoids growing (5-6 days) in matrigel I think that 1 um resolution will work for our projects. I am investigating systems from the following companies: Zeiss, Leica, La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase View. I think that is an exhaustive list of the commercial systems available but if you know of more, please let me know. I am fishing for feedback about hardware/software/data processing so that I can be more aware of what to look out for during demos. Thanks in advance to everyone who will contribute, online or offline alike. :) Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader, PhD Live Cell Imaging Facility Manager Karolinska Institutet- Bionut Dpt Hälsovägen 7, Novum, G lift, floor 6 14157 Huddinge Sweden mobile: +46 (0) 73 733 5008 office: +46 (0) 8 524 811 72 LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi Sylvie, CLARITY is sooooooo 2013 (and 2014 and 2015 and 2016). Ron Germain and colleagues recently published Ce3D (aka 3D HistoCytometry) http://www.pnas.org/content/114/35/E7321.abstract appendix includes on page 21 comparison table of lots of tissue clearing methods, http://www.pnas.org/content/suppl/2017/08/08/1708981114.DCSupplemental/pnas.1708981114.sapp.pdf Mostly used a confocal microscope. Consumables approach - sideview plates available in France, http://www.viewpoint.fr/en/p/equipment/sideview-microplate Publication: Rovira et al 2011 PNAS http://www.pnas.org/content/108/48/19264.long Another vendor: leica DLS with mirrors Leica SP8 confocal microscope http://www.leica-microsystems.com/products/confocal-microscopes/details/product/leica-tcs-sp8-dls/ The Olympus MacroView Zoom MVX10 is the basis for at least one of the light sheet products, https://www.olympus-lifescience.com/en/microscopes/macro/mvx10/ very cool MVPLAPO 2 XC objective lens available, 2x mag, 0.5 NA ... better specifications than Brad Amos Mesolens, http://www.mesolens.com/product/technical-specifications/ you could use the 2x/0.5NA with a pair of Cairn Researc L-SPI's to excite all four views. https://www.cairn-research.co.uk/product/lightsheet Disclosure: I manage an imaging core that hosts an Olympus Imaging Center http://confocal.jhu.edu/gallery George On 9/21/2017 3:24 AM, Sylvie Le Guyader wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Dear list > > We are currently preparing to tender for a light sheet system for our imaging facility. > The aim is to image 2 types of samples: > > - Tissue chunks, cleared with Clarity (mostly Fructose) and fluorescent, max @ 3x3x3 mm > > - Live organoids growing (5-6 days) in matrigel > > I think that 1 um resolution will work for our projects. > > I am investigating systems from the following companies: Zeiss, Leica, La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase View. I think that is an exhaustive list of the commercial systems available but if you know of more, please let me know. > > I am fishing for feedback about hardware/software/data processing so that I can be more aware of what to look out for during demos. Thanks in advance to everyone who will contribute, online or offline alike. :) > > Med vänlig hälsning / Best regards > > Sylvie > > @@@@@@@@@@@@@@@@@@@@@@@@ > Sylvie Le Guyader, PhD > Live Cell Imaging Facility Manager > Karolinska Institutet- Bionut Dpt > Hälsovägen 7, > Novum, G lift, floor 6 > 14157 Huddinge > Sweden > mobile: +46 (0) 73 733 5008 > office: +46 (0) 8 524 811 72 > LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> -- George McNamara, PhD Baltimore, MD 21231 [hidden email] https://www.linkedin.com/in/georgemcnamara https://works.bepress.com/gmcnamara/75 (may need to use Microsoft Edge or Firefox, rather than Google Chrome) http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650 http://confocal.jhu.edu July 2017 Current Protocols article, open access: UNIT 4.4 Microscopy and Image Analysis http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/abstract supporting materials direct link is http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/full#hg0404-sec-0023 figures at http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/figures |
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In reply to this post by Sylvie Le Guyader
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi Sylvie, Here is a summary of what was posted a few months ago on this list, nearly entirely compiled by Michael Weber: - Zeiss Lightsheet Z1 - Leica SP8 DLS - ASI diSPIM - ASI oSPIM - 3I Marianas - 3I Lattice Lightsheet - LaVision Ultramicroscope - Cairn L-SPI Illuminator - Luxendo MuVi-SPIM - Luxendo InVi-SPIM - Phaseview Alpha3 - M Squared Lasers Those are the future setups that I know of (in case you have time to wait): - Zeiss Lattice - Leica SCAPE Luxendo has since been acquired by Bruker but is continuing to use its name. Best regards, Jim James R. Mansfield Global OEM Business Development Manager Cell +1-978-831-8799 Web andor.com -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Sylvie Le Guyader Sent: Thursday, September 21, 2017 9:24 AM To: [hidden email] Subject: light sheet feedback ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear list We are currently preparing to tender for a light sheet system for our imaging facility. The aim is to image 2 types of samples: - Tissue chunks, cleared with Clarity (mostly Fructose) and fluorescent, max @ 3x3x3 mm - Live organoids growing (5-6 days) in matrigel I think that 1 um resolution will work for our projects. I am investigating systems from the following companies: Zeiss, Leica, La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase View. I think that is an exhaustive list of the commercial systems available but if you know of more, please let me know. I am fishing for feedback about hardware/software/data processing so that I can be more aware of what to look out for during demos. Thanks in advance to everyone who will contribute, online or offline alike. :) Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader, PhD Live Cell Imaging Facility Manager Karolinska Institutet- Bionut Dpt Hälsovägen 7, Novum, G lift, floor 6 14157 Huddinge Sweden mobile: +46 (0) 73 733 5008 office: +46 (0) 8 524 811 72 LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Another LSFM not yet mentioned... http://planelight.net/index.php/technology/ Cheers, Mark ************************************************** Mark A. Sanders University of Minnesota Program Director Twin Cities Campus University Imaging Centers www.uic.umn.edu > On Sep 21, 2017, at 6:44 AM, Jim Mansfield <[hidden email]> wrote: > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Hi Sylvie, > > Here is a summary of what was posted a few months ago on this list, nearly > entirely compiled by Michael Weber: > > - Zeiss Lightsheet Z1 > - Leica SP8 DLS > - ASI diSPIM > - ASI oSPIM > - 3I Marianas > - 3I Lattice Lightsheet > - LaVision Ultramicroscope > - Cairn L-SPI Illuminator > - Luxendo MuVi-SPIM > - Luxendo InVi-SPIM > - Phaseview Alpha3 > - M Squared Lasers > > Those are the future setups that I know of (in case you have time to wait): > > - Zeiss Lattice > - Leica SCAPE > > Luxendo has since been acquired by Bruker but is continuing to use its name. > > Best regards, > > Jim > > James R. Mansfield > Global OEM Business Development Manager > > > > Cell +1-978-831-8799 > Web andor.com > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] On > Behalf Of Sylvie Le Guyader > Sent: Thursday, September 21, 2017 9:24 AM > To: [hidden email] > Subject: light sheet feedback > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Dear list > > We are currently preparing to tender for a light sheet system for our > imaging facility. > The aim is to image 2 types of samples: > > - Tissue chunks, cleared with Clarity (mostly Fructose) and > fluorescent, max @ 3x3x3 mm > > - Live organoids growing (5-6 days) in matrigel > > I think that 1 um resolution will work for our projects. > > I am investigating systems from the following companies: Zeiss, Leica, La > Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase > View. I think that is an exhaustive list of the commercial systems available > but if you know of more, please let me know. > > I am fishing for feedback about hardware/software/data processing so that I > can be more aware of what to look out for during demos. Thanks in advance to > everyone who will contribute, online or offline alike. :) > > Med vänlig hälsning / Best regards > > Sylvie > > @@@@@@@@@@@@@@@@@@@@@@@@ > Sylvie Le Guyader, PhD > Live Cell Imaging Facility Manager > Karolinska Institutet- Bionut Dpt > Hälsovägen 7, > Novum, G lift, floor 6 > 14157 Huddinge > Sweden > mobile: +46 (0) 73 733 5008 > office: +46 (0) 8 524 811 72 > LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Thanks! I will check them out. Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader, PhD Live Cell Imaging Facility Manager Karolinska Institutet- Bionut Dpt Hälsovägen 7, Novum, G lift, floor 6 14157 Huddinge Sweden mobile: +46 (0) 73 733 5008 office: +46 (0) 8 524 811 72 LCI website -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Mark Sanders Sent: den 21 september 2017 13:58 To: [hidden email] Subject: Re: light sheet feedback ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Another LSFM not yet mentioned... http://planelight.net/index.php/technology/ Cheers, Mark ************************************************** Mark A. Sanders University of Minnesota Program Director Twin Cities Campus University Imaging Centers www.uic.umn.edu > On Sep 21, 2017, at 6:44 AM, Jim Mansfield <[hidden email]> wrote: > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Hi Sylvie, > > Here is a summary of what was posted a few months ago on this list, > nearly entirely compiled by Michael Weber: > > - Zeiss Lightsheet Z1 > - Leica SP8 DLS > - ASI diSPIM > - ASI oSPIM > - 3I Marianas > - 3I Lattice Lightsheet > - LaVision Ultramicroscope > - Cairn L-SPI Illuminator > - Luxendo MuVi-SPIM > - Luxendo InVi-SPIM > - Phaseview Alpha3 > - M Squared Lasers > > Those are the future setups that I know of (in case you have time to wait): > > - Zeiss Lattice > - Leica SCAPE > > Luxendo has since been acquired by Bruker but is continuing to use its name. > > Best regards, > > Jim > > James R. Mansfield > Global OEM Business Development Manager > > > > Cell +1-978-831-8799 > Web andor.com > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of Sylvie Le > Guyader > Sent: Thursday, September 21, 2017 9:24 AM > To: [hidden email] > Subject: light sheet feedback > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Dear list > > We are currently preparing to tender for a light sheet system for our > imaging facility. > The aim is to image 2 types of samples: > > - Tissue chunks, cleared with Clarity (mostly Fructose) and > fluorescent, max @ 3x3x3 mm > > - Live organoids growing (5-6 days) in matrigel > > I think that 1 um resolution will work for our projects. > > I am investigating systems from the following companies: Zeiss, Leica, > La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared > and Phase View. I think that is an exhaustive list of the commercial > systems available but if you know of more, please let me know. > > I am fishing for feedback about hardware/software/data processing so > that I can be more aware of what to look out for during demos. Thanks > in advance to everyone who will contribute, online or offline alike. > :) > > Med vänlig hälsning / Best regards > > Sylvie > > @@@@@@@@@@@@@@@@@@@@@@@@ > Sylvie Le Guyader, PhD > Live Cell Imaging Facility Manager > Karolinska Institutet- Bionut Dpt > Hälsovägen 7, > Novum, G lift, floor 6 > 14157 Huddinge > Sweden > mobile: +46 (0) 73 733 5008 > office: +46 (0) 8 524 811 72 > LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> |
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In reply to this post by George McNamara
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Re clearing A quicker protocol finally using a non toxic high RI medium Klingberg A et al, J Am Soc Nephrol, 28, 452-459, 2017 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of George McNamara Sent: den 21 september 2017 13:37 To: [hidden email] Subject: Re: light sheet feedback ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi Sylvie, CLARITY is sooooooo 2013 (and 2014 and 2015 and 2016). Ron Germain and colleagues recently published Ce3D (aka 3D HistoCytometry) http://www.pnas.org/content/114/35/E7321.abstract appendix includes on page 21 comparison table of lots of tissue clearing methods, http://www.pnas.org/content/suppl/2017/08/08/1708981114.DCSupplemental/pnas.1708981114.sapp.pdf Mostly used a confocal microscope. Consumables approach - sideview plates available in France, http://www.viewpoint.fr/en/p/equipment/sideview-microplate Publication: Rovira et al 2011 PNAS http://www.pnas.org/content/108/48/19264.long Another vendor: leica DLS with mirrors Leica SP8 confocal microscope http://www.leica-microsystems.com/products/confocal-microscopes/details/product/leica-tcs-sp8-dls/ The Olympus MacroView Zoom MVX10 is the basis for at least one of the light sheet products, https://www.olympus-lifescience.com/en/microscopes/macro/mvx10/ very cool MVPLAPO 2 XC objective lens available, 2x mag, 0.5 NA ... better specifications than Brad Amos Mesolens, http://www.mesolens.com/product/technical-specifications/ you could use the 2x/0.5NA with a pair of Cairn Researc L-SPI's to excite all four views. https://www.cairn-research.co.uk/product/lightsheet Disclosure: I manage an imaging core that hosts an Olympus Imaging Center http://confocal.jhu.edu/gallery George On 9/21/2017 3:24 AM, Sylvie Le Guyader wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Dear list > > We are currently preparing to tender for a light sheet system for our imaging facility. > The aim is to image 2 types of samples: > > - Tissue chunks, cleared with Clarity (mostly Fructose) and fluorescent, max @ 3x3x3 mm > > - Live organoids growing (5-6 days) in matrigel > > I think that 1 um resolution will work for our projects. > > I am investigating systems from the following companies: Zeiss, Leica, La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase View. I think that is an exhaustive list of the commercial systems available but if you know of more, please let me know. > > I am fishing for feedback about hardware/software/data processing so that I can be more aware of what to look out for during demos. Thanks in advance to everyone who will contribute, online or offline alike. :) > > Med vänlig hälsning / Best regards > > Sylvie > > @@@@@@@@@@@@@@@@@@@@@@@@ > Sylvie Le Guyader, PhD > Live Cell Imaging Facility Manager > Karolinska Institutet- Bionut Dpt > Hälsovägen 7, > Novum, G lift, floor 6 > 14157 Huddinge > Sweden > mobile: +46 (0) 73 733 5008 > office: +46 (0) 8 524 811 72 > LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> -- George McNamara, PhD Baltimore, MD 21231 [hidden email] https://www.linkedin.com/in/georgemcnamara https://works.bepress.com/gmcnamara/75 (may need to use Microsoft Edge or Firefox, rather than Google Chrome) http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650 http://confocal.jhu.edu July 2017 Current Protocols article, open access: UNIT 4.4 Microscopy and Image Analysis http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/abstract supporting materials direct link is http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/full#hg0404-sec-0023 figures at http://onlinelibrary.wiley.com/doi/10.1002/cphg.42/figures |
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In reply to this post by Sylvie Le Guyader
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Sylvie, Light sheet setups I’d take a closer look at for your applications are the Zeiss Lightsheet Z.1, the Luxendo MuVi-SPIM and the ASI diSPIM. If you want to image entire organoids you most likely want to have a classic multi view SPIM setup, e.g. the Z.1 or the MuVi-SPIM. The MuVi-SPIM does have the edge here because of the two detection arms which reduce the number of rotations required to get full 360 deg coverage. The diSPIM provides only two perpendicular views but might have an advantage for sample prep: you can line up many organoids and image them over time, very similar to an upright microscope. As far as I know, the diSPIM should support 10x dipping objectives that would give you a decent field of view for organoids. You have to get more creative with sample mounting for organoids in the classic SPIM configurations, e.g. using FEP tubes. I’m not up to date with all the clearing options, specifically the relevant objectives each setup supports. On the Zeiss you can use a low magnification air lens and an enclosed chamber, but I don’t know how compatible the MuVi-SPIM and the diSPIM would be for imaging of cleared samples. I definitely recommend organizing a demo for each of the systems you’re interested in. Best, Michael On Sep 21, 2017, at 3:24 AM, Sylvie Le Guyader <[hidden email]<mailto:[hidden email]>> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear list We are currently preparing to tender for a light sheet system for our imaging facility. The aim is to image 2 types of samples: - Tissue chunks, cleared with Clarity (mostly Fructose) and fluorescent, max @ 3x3x3 mm - Live organoids growing (5-6 days) in matrigel I think that 1 um resolution will work for our projects. I am investigating systems from the following companies: Zeiss, Leica, La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase View. I think that is an exhaustive list of the commercial systems available but if you know of more, please let me know. I am fishing for feedback about hardware/software/data processing so that I can be more aware of what to look out for during demos. Thanks in advance to everyone who will contribute, online or offline alike. :) Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader, PhD Live Cell Imaging Facility Manager Karolinska Institutet- Bionut Dpt Hälsovägen 7, Novum, G lift, floor 6 14157 Huddinge Sweden mobile: +46 (0) 73 733 5008 office: +46 (0) 8 524 811 72 LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> _____________ Dr. Michael Weber Advanced Microscopy Fellow Harvard Medical School 240 Longwood Ave, LHRRB 113, Boston, MA 02115 phone: +1 (617) 335-8395 http://nic.med.harvard.edu |
 
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Re: light sheet feedback
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Sylvie, I would like to add our QLS-scope Light Sheet Microscope into the candidates list that Michael suggested to you. Our QLS-scope is the only Light Sheet in the market with a patented technology of moving the light sheet through the sample and not the sample through the sheet. This gives us great benefits: 1. Speed. We can scan a whole 1 cm brain with 1000 stacks in 1.5 minutes. We can scan in 3D a whole zebra fish heart in the time of one heart beat. 2. Size. We can scan any sample, small and live, big and cleared. Using our stitching capacity we can scan samples much bigger than the field of view of any objective. 3. Resolution. We have the highest z resolution (around 1 micron). Measuring at 0 and 180 degrees and reconstructing the image we provide 3D images of a big sample with the same resolution close and far away from the camera. Measuring at 4 or more angles (up to 500) and reconstruction the 3D image we can demonstrate isotropic resolution in big cleared samples. The 3D reconstruction of a SPIM measurement, stitching and several angles 3D reconstruction is done automatically by our software. These and much more advantages can be found in our webpage: www.planelight.net We can run any of your samples any time you wish and since the system is so fast we can do it on-line so you can see how we measure as well as the results in real time. Best regards Nikos Ekizoglou Plane Light S.L. C/ Riocabado, 4 28047 Madrid (Spain) Phone: +34 911 130 824 Cell: +34 650 70 52 39 Fax: +34 910 113 757 www.planelight.net -----Mensaje original----- De: Confocal Microscopy List [mailto:[hidden email]] En nombre de Weber, Michael Enviado el: viernes, 29 de septiembre de 2017 19:20 Para: [hidden email] Asunto: Re: light sheet feedback ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Sylvie, Light sheet setups I’d take a closer look at for your applications are the Zeiss Lightsheet Z.1, the Luxendo MuVi-SPIM and the ASI diSPIM. If you want to image entire organoids you most likely want to have a classic multi view SPIM setup, e.g. the Z.1 or the MuVi-SPIM. The MuVi-SPIM does have the edge here because of the two detection arms which reduce the number of rotations required to get full 360 deg coverage. The diSPIM provides only two perpendicular views but might have an advantage for sample prep: you can line up many organoids and image them over time, very similar to an upright microscope. As far as I know, the diSPIM should support 10x dipping objectives that would give you a decent field of view for organoids. You have to get more creative with sample mounting for organoids in the classic SPIM configurations, e.g. using FEP tubes. I’m not up to date with all the clearing options, specifically the relevant objectives each setup supports. On the Zeiss you can use a low magnification air lens and an enclosed chamber, but I don’t know how compatible the MuVi-SPIM and the diSPIM would be for imaging of cleared samples. I definitely recommend organizing a demo for each of the systems you’re interested in. Best, Michael On Sep 21, 2017, at 3:24 AM, Sylvie Le Guyader <[hidden email]<mailto:[hidden email]>> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear list We are currently preparing to tender for a light sheet system for our imaging facility. The aim is to image 2 types of samples: - Tissue chunks, cleared with Clarity (mostly Fructose) and fluorescent, max @ 3x3x3 mm - Live organoids growing (5-6 days) in matrigel I think that 1 um resolution will work for our projects. I am investigating systems from the following companies: Zeiss, Leica, La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase View. I think that is an exhaustive list of the commercial systems available but if you know of more, please let me know. I am fishing for feedback about hardware/software/data processing so that I can be more aware of what to look out for during demos. Thanks in advance to everyone who will contribute, online or offline alike. :) Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader, PhD Live Cell Imaging Facility Manager Karolinska Institutet- Bionut Dpt Hälsovägen 7, Novum, G lift, floor 6 14157 Huddinge Sweden mobile: +46 (0) 73 733 5008 office: +46 (0) 8 524 811 72 LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> _____________ Dr. Michael Weber Advanced Microscopy Fellow Harvard Medical School 240 Longwood Ave, LHRRB 113, Boston, MA 02115 phone: +1 (617) 335-8395 http://nic.med.harvard.edu |
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In reply to this post by Weber, Michael
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** In response to the ***commercial posting*** (though not marked as such) below: Very interesting... was mentioned on the list few weeks ago. Since you're advertising to the community, I have some questions: "Moving LightSheet instead of the sample ..." means you have to move the objective lens as well (unless you're using some sort of remote focusing, but that does not cover large volumes), right? Can you share some info on how you do that exactly? How do you correct for the varying spherical aberration with dry lenses? And how do you make this work with immersion lenses? All papers on your website refer to the traditional approach (moving sample) with a cylindrical lens; one mentions "adapting a conventional digitally- scanned light sheet fluorescence microscope". Which brings another question: how do you achieve the claimed 1 um lightsheet thickness? Over what field? The practical values mentioned in the papers are quite a bit higher... The cylindrical-lens approach is mentioned on your website, and there is little room for adaptive optics to bring any improvements... And what CMOS camera are you using (with < 1 e- read noise)? I know that Andor (for example) claims such values, but it's at lower speed and more importantly, it's median values, while other vendors use RMS noise (seems more appropriate to me, and the values are, as far as I know, > 1e-)... Finally, when it's going to be available? Any demos? Your colleagues at 4d- nature mention that you "will commercialize...", has that already happened? Btw, I've noticed some strange values of FOV for some of the listed objectives (compare 10x dry: 2.2mm FOV, vs 10x water: 20 mm FOV), is that correct? also "Tweeter" is not the correct spelling of the service :-). Good luck with your product! zdenek -- Zdenek Svindrych, Ph.D. W.M. Keck Center for Cellular Imaging (PLSB 003) Department of Biology,University of Virginia 409 McCormick Rd, Charlottesville, VA-22904 http://www.kcci.virginia.edu/ tel: 434-982-4869 ---------- Původní e-mail ---------- Od: Nikos Ekizoglou - Planelight <[hidden email]> Komu: [hidden email] Datum: 2. 10. 2017 7:10:00 Předmět: Re: light sheet feedback "***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Sylvie, I would like to add our QLS-scope Light Sheet Microscope into the candidates list that Michael suggested to you. Our QLS-scope is the only Light Sheet in the market with a patented technology of moving the light sheet through the sample and not the sample through the sheet. This gives us great benefits: 1. Speed. We can scan a whole 1 cm brain with 1000 stacks in 1.5 minutes. We can scan in 3D a whole zebra fish heart in the time of one heart beat. 2. Size. We can scan any sample, small and live, big and cleared. Using our stitching capacity we can scan samples much bigger than the field of view of any objective. 3. Resolution. We have the highest z resolution (around 1 micron). Measuring at 0 and 180 degrees and reconstructing the image we provide 3D images of a big sample with the same resolution close and far away from the camera. Measuring at 4 or more angles (up to 500) and reconstruction the 3D image we can demonstrate isotropic resolution in big cleared samples. The 3D reconstruction of a SPIM measurement, stitching and several angles 3D reconstruction is done automatically by our software. These and much more advantages can be found in our webpage: www.planelight. net We can run any of your samples any time you wish and since the system is so fast we can do it on-line so you can see how we measure as well as the results in real time. Best regards Nikos Ekizoglou Plane Light S.L. C/ Riocabado, 4 28047 Madrid (Spain) Phone: +34 911 130 824 Cell: +34 650 70 52 39 Fax: +34 910 113 757 www.planelight.net -----Mensaje original----- De: Confocal Microscopy List [mailto:[hidden email]] En nombre de Weber, Michael Enviado el: viernes, 29 de septiembre de 2017 19:20 Para: [hidden email] Asunto: Re: light sheet feedback ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Sylvie, Light sheet setups I’d take a closer look at for your applications are the Zeiss Lightsheet Z.1, the Luxendo MuVi-SPIM and the ASI diSPIM. If you want to image entire organoids you most likely want to have a classic multi view SPIM setup, e.g. the Z.1 or the MuVi-SPIM. The MuVi-SPIM does have the edge here because of the two detection arms which reduce the number of rotations required to get full 360 deg coverage. The diSPIM provides only two perpendicular views but might have an advantage for sample prep: you can line up many organoids and image them over time, very similar to an upright microscope. As far as I know, the diSPIM should support 10x dipping objectives that would give you a decent field of view for organoids. You have to get more creative with sample mounting for organoids in the classic SPIM configurations, e.g. using FEP tubes. I’m not up to date with all the clearing options, specifically the relevant objectives each setup supports. On the Zeiss you can use a low magnification air lens and an enclosed chamber, but I don’t know how compatible the MuVi-SPIM and the diSPIM would be for imaging of cleared samples. I definitely recommend organizing a demo for each of the systems you’re interested in. Best, Michael On Sep 21, 2017, at 3:24 AM, Sylvie Le Guyader <[hidden email]< mailto:[hidden email]>> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear list We are currently preparing to tender for a light sheet system for our imaging facility. The aim is to image 2 types of samples: - Tissue chunks, cleared with Clarity (mostly Fructose) and fluorescent, max @ 3x3x3 mm - Live organoids growing (5-6 days) in matrigel I think that 1 um resolution will work for our projects. I am investigating systems from the following companies: Zeiss, Leica, La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase View. I think that is an exhaustive list of the commercial systems available but if you know of more, please let me know. I am fishing for feedback about hardware/software/data processing so that I can be more aware of what to look out for during demos. Thanks in advance to everyone who will contribute, online or offline alike. :) Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader, PhD Live Cell Imaging Facility Manager Karolinska Institutet- Bionut Dpt Hälsovägen 7, Novum, G lift, floor 6 14157 Huddinge Sweden mobile: +46 (0) 73 733 5008 office: +46 (0) 8 524 811 72 LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> _____________ Dr. Michael Weber Advanced Microscopy Fellow Harvard Medical School 240 Longwood Ave, LHRRB 113, Boston, MA 02115 phone: +1 (617) 335-8395 http://nic.med.harvard.edu " |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** On Mon, Oct 2, 2017 at 11:08 AM, <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > > "Moving LightSheet instead of the sample ..." means you have to move the > objective lens as well (unless you're using some sort of remote focusing, > but that does not cover large volumes), right? Can you share some info on > how you do that exactly? How do you correct for the varying spherical > aberration with dry lenses? And how do you make this work with immersion > lenses? > > The version I've seen moves the sheet in the focus using a galvo mirror. in at an oblique angle, rather like TIRF. As to how the vendor maintains thickness etc. they will have to answer that themselves. :) Craig |
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Nikos Ekizoglou - Planelight |
Re: light sheet feedback***commercial posting***
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In reply to this post by Zdenek Svindrych-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 Post images on http://www.imgur.com and include the link in your posting. ***** Dear Zdenek, I have already apologized to the list owner about the post. We just joined the list so I was not aware of all the rules. Now regarding your questions: "Moving LightSheet instead of the sample ..." means you have to move the objective lens as well (unless you're using some sort of remote focusing, but that does not cover large volumes), right? Can you share some info on how you do that exactly? How do you correct for the varying spherical aberration with dry lenses? And how do you make this work with immersion lenses? What you mention is one of the ways used in light sheet microscopy, but not the only one. Another approach is to use a tunable lens in the detection arm, thus removing the need to move the objective (either an acoustic tunable lens or an electric tunable lens). If the light sheet is then scanned using galvos, there is no need to move the sample. This issue then is linked to your next question, how do we correct for the spherical aberration: the answer is that we don´t, at least at the moment. For the time being we have concentrated in imaging at the mesoscale, which is where there is a lack of imaging modalities. Our system images samples from 2cm to 1mm in diameter by using magnifications between 1x and 20x (water). From our experience, using up to 20x (water) we do not see an important contribution of spherical aberration, at least for in-vivo imaging. However, we do have custom light sheet system that corrects for aberrations using a LCOS, but we have so far implemented it on the illumination arm only, to ensure a thin and maintained light sheet even in the presence of scattering. A similar approach could be used for the detection arm. This custom light sheet was aimed at imaging at higher resolution, using objectives of 40x or above. Note that in this case imaging would be extremely slow, and speed is one of our main goals, which is why it is not currently implemented in the Qls-scope. All papers on your website refer to the traditional approach (moving sample) with a cylindrical lens; one mentions "adapting a conventional digitally- scanned light sheet fluorescence microscope". Which brings another question: First of all we need to clarify that almost none of the technology that we have implemented in the Qls-scope has been published, we are at the moment drafting a few papers on the subject. Hence, the papers you have seen are indeed using the first versions of the light sheet microscope were we did indeed move the sample. Having said this, here we answer your questions related to resolution: how do you achieve the claimed 1 um lightsheet thickness? Over what field? One of the advantages of working with large samples is that we can scale our imaging system appropriately. As you are obviously aware, what we ideally want are high numerical aperture lenses and long working distances which will in turn reduce the field of view over which the light sheet is as thin as possible (note that we use a gaussian beam). Our approach is to avoid using an objective for illumination, and thus in turn use a set of large diameter lenses which effectively improve the NA and due to their size still maintain a long working distance (compared to the sample size). However, the optimal width of our beam is only present in a portion of the image (depending on the imaging objective). We deal with this issue by using an extra tunable lens that sweeps in a very controlled fashion over the field of view. The practical values mentioned in the papers are quite a bit higher... The cylindrical-lens approach is mentioned on your website, and there is little room for adaptive optics to bring any improvements... You are correct, as mentioned previously those papers were based on a much older setup. As mentioned above, we did implement an LCOS to improve the sheet in the presence of scattering. We have yet to implement such an approach on the detection arm, but note that for example Pablo Loza at ICFO has achieved this, so it is feasible. Note that reducing the speed of acquisition though is something we want to avoid unless it is absolutely necessary. And what CMOS camera are you using (with < 1 e- read noise)? I know that Andor (for example) claims such values, but it's at lower speed and more importantly, it's median values, while other vendors use RMS noise (seems more appropriate to me, and the values are, as far as I know, > 1e-)... We use Hamamatsu, in particular the one implemented now is the Orca Flash-4.0 R2 with cameralink. You might find more info here: http://camera.hamamatsu.com/jp/en/product/search/C13440-20CU/index.html Finally, when it's going to be available? Any demos? Your colleagues at 4d- nature mention that you "will commercialize...", has that already happened? The system is already available. We have already performed demos at customer sites in Europe and we are programming webinars where we can show the system working live. It takes very little time to measure even a very big sample so it is feasible. If you want a live demonstration for you and the people from your lab just let us know. Btw, I've noticed some strange values of FOV for some of the listed objectives (compare 10x dry: 2.2mm FOV, vs 10x water: 20 mm FOV), is that correct? also "Tweeter" is not the correct spelling of the service :-). We can implement any available objective in the market. We have found some of the best air objectives in the market that are specifically designed with high N/A. We also use Zeiss, Leica or Olympus objectives for immersion. We really can use any objective fits better to the researcher application. Good luck with your product! THANKS! -----Mensaje original----- De: Confocal Microscopy List [mailto:[hidden email]] En nombre de [hidden email] Enviado el: lunes, 02 de octubre de 2017 19:09 Para: [hidden email] Asunto: Re: light sheet feedback ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** In response to the ***commercial posting*** (though not marked as such) below: Very interesting... was mentioned on the list few weeks ago. Since you're advertising to the community, I have some questions: "Moving LightSheet instead of the sample ..." means you have to move the objective lens as well (unless you're using some sort of remote focusing, but that does not cover large volumes), right? Can you share some info on how you do that exactly? How do you correct for the varying spherical aberration with dry lenses? And how do you make this work with immersion lenses? All papers on your website refer to the traditional approach (moving sample) with a cylindrical lens; one mentions "adapting a conventional digitally- scanned light sheet fluorescence microscope". Which brings another question: how do you achieve the claimed 1 um lightsheet thickness? Over what field? The practical values mentioned in the papers are quite a bit higher... The cylindrical-lens approach is mentioned on your website, and there is little room for adaptive optics to bring any improvements... And what CMOS camera are you using (with < 1 e- read noise)? I know that Andor (for example) claims such values, but it's at lower speed and more importantly, it's median values, while other vendors use RMS noise (seems more appropriate to me, and the values are, as far as I know, > 1e-)... Finally, when it's going to be available? Any demos? Your colleagues at 4d- nature mention that you "will commercialize...", has that already happened? Btw, I've noticed some strange values of FOV for some of the listed objectives (compare 10x dry: 2.2mm FOV, vs 10x water: 20 mm FOV), is that correct? also "Tweeter" is not the correct spelling of the service :-). Good luck with your product! zdenek -- Zdenek Svindrych, Ph.D. W.M. Keck Center for Cellular Imaging (PLSB 003) Department of Biology,University of Virginia 409 McCormick Rd, Charlottesville, VA-22904 http://www.kcci.virginia.edu/ tel: 434-982-4869 ---------- Původní e-mail ---------- Od: Nikos Ekizoglou - Planelight <[hidden email]> Komu: [hidden email] Datum: 2. 10. 2017 7:10:00 Předmět: Re: light sheet feedback "***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Sylvie, I would like to add our QLS-scope Light Sheet Microscope into the candidates list that Michael suggested to you. Our QLS-scope is the only Light Sheet in the market with a patented technology of moving the light sheet through the sample and not the sample through the sheet. This gives us great benefits: 1. Speed. We can scan a whole 1 cm brain with 1000 stacks in 1.5 minutes. We can scan in 3D a whole zebra fish heart in the time of one heart beat. 2. Size. We can scan any sample, small and live, big and cleared. Using our stitching capacity we can scan samples much bigger than the field of view of any objective. 3. Resolution. We have the highest z resolution (around 1 micron). Measuring at 0 and 180 degrees and reconstructing the image we provide 3D images of a big sample with the same resolution close and far away from the camera. Measuring at 4 or more angles (up to 500) and reconstruction the 3D image we can demonstrate isotropic resolution in big cleared samples. The 3D reconstruction of a SPIM measurement, stitching and several angles 3D reconstruction is done automatically by our software. These and much more advantages can be found in our webpage: www.planelight. net We can run any of your samples any time you wish and since the system is so fast we can do it on-line so you can see how we measure as well as the results in real time. Best regards Nikos Ekizoglou Plane Light S.L. C/ Riocabado, 4 28047 Madrid (Spain) Phone: +34 911 130 824 Cell: +34 650 70 52 39 Fax: +34 910 113 757 www.planelight.net -----Mensaje original----- De: Confocal Microscopy List [mailto:[hidden email]] En nombre de Weber, Michael Enviado el: viernes, 29 de septiembre de 2017 19:20 Para: [hidden email] Asunto: Re: light sheet feedback ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Sylvie, Light sheet setups I’d take a closer look at for your applications are the Zeiss Lightsheet Z.1, the Luxendo MuVi-SPIM and the ASI diSPIM. If you want to image entire organoids you most likely want to have a classic multi view SPIM setup, e.g. the Z.1 or the MuVi-SPIM. The MuVi-SPIM does have the edge here because of the two detection arms which reduce the number of rotations required to get full 360 deg coverage. The diSPIM provides only two perpendicular views but might have an advantage for sample prep: you can line up many organoids and image them over time, very similar to an upright microscope. As far as I know, the diSPIM should support 10x dipping objectives that would give you a decent field of view for organoids. You have to get more creative with sample mounting for organoids in the classic SPIM configurations, e.g. using FEP tubes. I’m not up to date with all the clearing options, specifically the relevant objectives each setup supports. On the Zeiss you can use a low magnification air lens and an enclosed chamber, but I don’t know how compatible the MuVi-SPIM and the diSPIM would be for imaging of cleared samples. I definitely recommend organizing a demo for each of the systems you’re interested in. Best, Michael On Sep 21, 2017, at 3:24 AM, Sylvie Le Guyader <[hidden email]< mailto:[hidden email]>> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear list We are currently preparing to tender for a light sheet system for our imaging facility. The aim is to image 2 types of samples: - Tissue chunks, cleared with Clarity (mostly Fructose) and fluorescent, max @ 3x3x3 mm - Live organoids growing (5-6 days) in matrigel I think that 1 um resolution will work for our projects. I am investigating systems from the following companies: Zeiss, Leica, La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase View. I think that is an exhaustive list of the commercial systems available but if you know of more, please let me know. I am fishing for feedback about hardware/software/data processing so that I can be more aware of what to look out for during demos. Thanks in advance to everyone who will contribute, online or offline alike. :) Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader, PhD Live Cell Imaging Facility Manager Karolinska Institutet- Bionut Dpt Hälsovägen 7, Novum, G lift, floor 6 14157 Huddinge Sweden mobile: +46 (0) 73 733 5008 office: +46 (0) 8 524 811 72 LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> _____________ Dr. Michael Weber Advanced Microscopy Fellow Harvard Medical School 240 Longwood Ave, LHRRB 113, Boston, MA 02115 phone: +1 (617) 335-8395 http://nic.med.harvard.edu " |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi Nikos, thanks for your reply. It indeed seems you are combining some nice technology to get an edge. But there still are some loose ends... I'm only guessing you are actually using a scanned "pencil" beam, not a cylindrical lens, to create the lightsheet. Otherwise it's difficult to sweep the focus of the sheet (unless you use a tuneable cylindrical lens). Also you mentioned 'galvos', a plural :-). And if you sweep-focus a gaussian "pencil" beam, you get a lot of light outside the focus (just like with bessel beams), so you definitely want to use the camera rolling shutter to improve the contrast. But there are more ways to do that. I can't wait to see your papers published! Btw, what I meant with the huge FOVs of the 10x and 20x water lenses (and the Tweeter thing) is ... those are typos, you don't want that on your company website :-). Don't confuse field of view and field number. And of course, the FOV depends on your camera chip size and the (fixed) zoom of your system, not necessarily what the lens manufacturer says. But then, a Zeiss 10x lens will have very different magnification than Nikon 10x lens. So you may want to report the actual values, if you want to be "Quantitative". And that magnification will be different from what's written on the lens barrel, which may confuse the users... we're moving in a circle here, I know, but don't let the business strategy win over the science :-). Good luck! zdenek ---------- Původní e-mail ---------- Od: Nikos Ekizoglou - Planelight <[hidden email]> Komu: [hidden email] Datum: 3. 10. 2017 5:21:22 Předmět: Re: light sheet feedback***commercial posting*** "***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Zdenek, I have already apologized to the list owner about the post. We just joined the list so I was not aware of all the rules. Now regarding your questions: "Moving LightSheet instead of the sample ..." means you have to move the objective lens as well (unless you're using some sort of remote focusing, but that does not cover large volumes), right? Can you share some info on how you do that exactly? How do you correct for the varying spherical aberration with dry lenses? And how do you make this work with immersion lenses? What you mention is one of the ways used in light sheet microscopy, but not the only one. Another approach is to use a tunable lens in the detection arm, thus removing the need to move the objective (either an acoustic tunable lens or an electric tunable lens). If the light sheet is then scanned using galvos, there is no need to move the sample. This issue then is linked to your next question, how do we correct for the spherical aberration: the answer is that we don´t, at least at the moment. For the time being we have concentrated in imaging at the mesoscale, which is where there is a lack of imaging modalities. Our system images samples from 2cm to 1mm in diameter by using magnifications between 1x and 20x (water). From our experience, using up to 20x (water) we do not see an important contribution of spherical aberration, at least for in-vivo imaging. However, we do have custom light sheet system that corrects for aberrations using a LCOS, but we have so far implemented it on the illumination arm only, to ensure a thin and maintained light sheet even in the presence of scattering. A similar approach could be used for the detection arm. This custom light sheet was aimed at imaging at higher resolution, using objectives of 40x or above. Note that in this case imaging would be extremely slow, and speed is one of our main goals, which is why it is not currently implemented in the Qls- scope. All papers on your website refer to the traditional approach (moving sample) with a cylindrical lens; one mentions "adapting a conventional digitally- scanned light sheet fluorescence microscope". Which brings another question: First of all we need to clarify that almost none of the technology that we have implemented in the Qls-scope has been published, we are at the moment drafting a few papers on the subject. Hence, the papers you have seen are indeed using the first versions of the light sheet microscope were we did indeed move the sample. Having said this, here we answer your questions related to resolution: how do you achieve the claimed 1 um lightsheet thickness? Over what field? One of the advantages of working with large samples is that we can scale our imaging system appropriately. As you are obviously aware, what we ideally want are high numerical aperture lenses and long working distances which will in turn reduce the field of view over which the light sheet is as thin as possible (note that we use a gaussian beam). Our approach is to avoid using an objective for illumination, and thus in turn use a set of large diameter lenses which effectively improve the NA and due to their size still maintain a long working distance (compared to the sample size). However, the optimal width of our beam is only present in a portion of the image (depending on the imaging objective). We deal with this issue by using an extra tunable lens that sweeps in a very controlled fashion over the field of view. The practical values mentioned in the papers are quite a bit higher... The cylindrical-lens approach is mentioned on your website, and there is little room for adaptive optics to bring any improvements... You are correct, as mentioned previously those papers were based on a much older setup. As mentioned above, we did implement an LCOS to improve the sheet in the presence of scattering. We have yet to implement such an approach on the detection arm, but note that for example Pablo Loza at ICFO has achieved this, so it is feasible. Note that reducing the speed of acquisition though is something we want to avoid unless it is absolutely necessary. And what CMOS camera are you using (with < 1 e- read noise)? I know that Andor (for example) claims such values, but it's at lower speed and more importantly, it's median values, while other vendors use RMS noise (seems more appropriate to me, and the values are, as far as I know, > 1e-)... We use Hamamatsu, in particular the one implemented now is the Orca Flash- 4.0 R2 with cameralink. You might find more info here: http://camera.hamamatsu.com/jp/en/product/search/C13440-20CU/index.html Finally, when it's going to be available? Any demos? Your colleagues at 4d- nature mention that you "will commercialize...", has that already happened? The system is already available. We have already performed demos at customer sites in Europe and we are programming webinars where we can show the system working live. It takes very little time to measure even a very big sample so it is feasible. If you want a live demonstration for you and the people from your lab just let us know. Btw, I've noticed some strange values of FOV for some of the listed objectives (compare 10x dry: 2.2mm FOV, vs 10x water: 20 mm FOV), is that correct? also "Tweeter" is not the correct spelling of the service :-). We can implement any available objective in the market. We have found some of the best air objectives in the market that are specifically designed with high N/A. We also use Zeiss, Leica or Olympus objectives for immersion. We really can use any objective fits better to the researcher application. Good luck with your product! THANKS! -----Mensaje original----- De: Confocal Microscopy List [mailto:[hidden email]] En nombre de [hidden email] Enviado el: lunes, 02 de octubre de 2017 19:09 Para: [hidden email] Asunto: Re: light sheet feedback ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** In response to the ***commercial posting*** (though not marked as such) below: Very interesting... was mentioned on the list few weeks ago. Since you're advertising to the community, I have some questions: "Moving LightSheet instead of the sample ..." means you have to move the objective lens as well (unless you're using some sort of remote focusing, but that does not cover large volumes), right? Can you share some info on how you do that exactly? How do you correct for the varying spherical aberration with dry lenses? And how do you make this work with immersion lenses? All papers on your website refer to the traditional approach (moving sample) with a cylindrical lens; one mentions "adapting a conventional digitally- scanned light sheet fluorescence microscope". Which brings another question: how do you achieve the claimed 1 um lightsheet thickness? Over what field? The practical values mentioned in the papers are quite a bit higher... The cylindrical-lens approach is mentioned on your website, and there is little room for adaptive optics to bring any improvements... And what CMOS camera are you using (with < 1 e- read noise)? I know that Andor (for example) claims such values, but it's at lower speed and more importantly, it's median values, while other vendors use RMS noise (seems more appropriate to me, and the values are, as far as I know, > 1e-)... Finally, when it's going to be available? Any demos? Your colleagues at 4d- nature mention that you "will commercialize...", has that already happened? Btw, I've noticed some strange values of FOV for some of the listed objectives (compare 10x dry: 2.2mm FOV, vs 10x water: 20 mm FOV), is that correct? also "Tweeter" is not the correct spelling of the service :-). Good luck with your product! zdenek -- Zdenek Svindrych, Ph.D. W.M. Keck Center for Cellular Imaging (PLSB 003) Department of Biology, University of Virginia 409 McCormick Rd, Charlottesville, VA-22904 http://www.kcci.virginia.edu/ tel: 434-982-4869 ---------- Původní e-mail ---------- Od: Nikos Ekizoglou - Planelight <[hidden email]> Komu: [hidden email] Datum: 2. 10. 2017 7:10:00 Předmět: Re: light sheet feedback "***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Sylvie, I would like to add our QLS-scope Light Sheet Microscope into the candidates list that Michael suggested to you. Our QLS-scope is the only Light Sheet in the market with a patented technology of moving the light sheet through the sample and not the sample through the sheet. This gives us great benefits: 1. Speed. We can scan a whole 1 cm brain with 1000 stacks in 1.5 minutes. We can scan in 3D a whole zebra fish heart in the time of one heart beat. 2. Size. We can scan any sample, small and live, big and cleared. Using our stitching capacity we can scan samples much bigger than the field of view of any objective. 3. Resolution. We have the highest z resolution (around 1 micron). Measuring at 0 and 180 degrees and reconstructing the image we provide 3D images of a big sample with the same resolution close and far away from the camera. Measuring at 4 or more angles (up to 500) and reconstruction the 3D image we can demonstrate isotropic resolution in big cleared samples. The 3D reconstruction of a SPIM measurement, stitching and several angles 3D reconstruction is done automatically by our software. These and much more advantages can be found in our webpage: www.planelight. net We can run any of your samples any time you wish and since the system is so fast we can do it on-line so you can see how we measure as well as the results in real time. Best regards Nikos Ekizoglou Plane Light S.L. C/ Riocabado, 4 28047 Madrid (Spain) Phone: +34 911 130 824 Cell: +34 650 70 52 39 Fax: +34 910 113 757 www.planelight.net -----Mensaje original----- De: Confocal Microscopy List [mailto:[hidden email]] En nombre de Weber, Michael Enviado el: viernes, 29 de septiembre de 2017 19:20 Para: [hidden email] Asunto: Re: light sheet feedback ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Sylvie, Light sheet setups I’d take a closer look at for your applications are the Zeiss Lightsheet Z.1, the Luxendo MuVi-SPIM and the ASI diSPIM. If you want to image entire organoids you most likely want to have a classic multi view SPIM setup, e.g. the Z.1 or the MuVi-SPIM. The MuVi-SPIM does have the edge here because of the two detection arms which reduce the number of rotations required to get full 360 deg coverage. The diSPIM provides only two perpendicular views but might have an advantage for sample prep: you can line up many organoids and image them over time, very similar to an upright microscope. As far as I know, the diSPIM should support 10x dipping objectives that would give you a decent field of view for organoids. You have to get more creative with sample mounting for organoids in the classic SPIM configurations, e.g. using FEP tubes. I’m not up to date with all the clearing options, specifically the relevant objectives each setup supports. On the Zeiss you can use a low magnification air lens and an enclosed chamber, but I don’t know how compatible the MuVi-SPIM and the diSPIM would be for imaging of cleared samples. I definitely recommend organizing a demo for each of the systems you’re interested in. Best, Michael On Sep 21, 2017, at 3:24 AM, Sylvie Le Guyader <[hidden email]< mailto:[hidden email]>> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear list We are currently preparing to tender for a light sheet system for our imaging facility. The aim is to image 2 types of samples: - Tissue chunks, cleared with Clarity (mostly Fructose) and fluorescent, max @ 3x3x3 mm - Live organoids growing (5-6 days) in matrigel I think that 1 um resolution will work for our projects. I am investigating systems from the following companies: Zeiss, Leica, La Vision Biotech, ASI, 3i/ASI, Luxendo, Cairns research, M Squared and Phase View. I think that is an exhaustive list of the commercial systems available but if you know of more, please let me know. I am fishing for feedback about hardware/software/data processing so that I can be more aware of what to look out for during demos. Thanks in advance to everyone who will contribute, online or offline alike. :) Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader, PhD Live Cell Imaging Facility Manager Karolinska Institutet- Bionut Dpt Hälsovägen 7, Novum, G lift, floor 6 14157 Huddinge Sweden mobile: +46 (0) 73 733 5008 office: +46 (0) 8 524 811 72 LCI website<http://ki.se/en/bionut/welcome-to-the-lci-facility> _____________ Dr. Michael Weber Advanced Microscopy Fellow Harvard Medical School 240 Longwood Ave, LHRRB 113, Boston, MA 02115 phone: +1 (617) 335-8395 http://nic.med.harvard.edu " " |
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