*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Since the topic of home built super-resolution systems has been brought up I was wondering if anyone had a very simple "Super Res for Dummies" type manual for building such a system. While I feel quite capable of following directions and assembling components together I have a certain degree of trepidation when it comes to figuring out how to drive all the components software-wise. I will admit I lack programming experience and limit myself to making ImageJ macros (poorly) or Cell Profiler pipelines - not writing code in Matlab to open shutters and such. I am aware that many articles describe their setups and give overview diagrams but, from a true schematic standpoint, they lack sufficient detail for someone as rigidly OCD as myself. Perhaps there is a class or course offered somewhere that covers this "roll your own" approach and I have yet to convince Google to divulge that information. Along the same lines, I am very interested in trying to construct an optical projection tomography system for our facility and, again, while I know many labs have built their own systems and published details of them, some critical details elude me. Have any listers built their own OPT rig and, if so, could you provide a detailed component list as well as assembly instructions? Personally, I would be more than willing to come visit a lab that has implemented their own solution to the FPALM/STORM and/or OPT method if they wouldn't mind spending a little time answering what they most likely would think are silly questions (and would allow me to take pictures of their system). David David H. Burk, PhD Cell Biology and Bioimaging Core Pennington Biomedical Research Center Baton Rouge, LA 70808 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Alberto Diaspro Sent: Saturday, January 29, 2011 10:47 AM To: [hidden email] Subject: Re: Who has purchased a fluorescence nanoscope? ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** At the Italian Institute of Technology, we got a Leica STED-CW and home built an FPALM on an inverted Nikon thanks to software sharing from Sam Hess Lab, University of Maine. We home built a WLL and a CW STED controlled by MPI Nanobiophotonics software. We are currently interested n the Nikon N-STORM. All the best Alby Il giorno 29/gen/2011, alle ore 17.19, Christophe Leterrier ha scritto: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > The Bordeaux Imaging Center in Bordeaux, France has a STED and a > TIRF/PALM http://www.bic.u-bordeaux2.fr/ > > PALM/STORM setups are becoming more common (I'm aware of 2 setups > already running in Marseille), because (at least this is what the > people who did it told me) it is quite easy to add to an existing TIRF > setup (provided you find software for the detection and localisation > of individual fluorophores, but there is now an available ImageJ plugin for that). > > > -- > Christophe Leterrier > Postdoc > INSERM UMR641 // Ionic channels Lab > IFR Jean Roche, Mediterranée University Marseille, France > [hidden email] > > > > > On Sat, Jan 29, 2011 at 16:52, G. Esteban Fernandez < > [hidden email]> wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> UCLA has a STED >> >> http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791 >> <http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791> >> http://www.cnsi.ucla.edu/staticpages/core-facilities#alms >> >> >> <http://www.cnsi.ucla.edu/staticpages/core-facilities#alms> >> >> On Sat, Jan 29, 2011 at 7:32 AM, George McNamara >> <[hidden email]>wrote: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> Dear Confocal listserv, >>> >>> Who has purchased a fluorescence nanoscope? What can you tell me >>> about >> your >>> experiences - either here or privately? >>> >>> I see 11 OMX labs listed at http://www.api.com/omx-labs.asp (may be >>> out >> of >>> date or some customers may be shy). >>> >>> I am aware of STED systems in the USA at Yale Univ, NIH and I was >>> told >> San >>> Diego (have not found where), and possibly UC Denver. Paul French >> apparently >>> did his own upgrade of a Leica SP2 ( >>> http://www.focusonmicroscopy.org/2008/PDF/070_Auksorius.pdf and a >>> Leica Scientific Forum video ... see also related work at >>> ftp://ftp.espci.fr/incoming/Gilles/INSERM-STED-x6.pdf ). >>> >>> I am also aware of one 4pi microscope in the USA. >>> >>> So, who has N-SIM, N-STORM, SR-SIM, PALm, SR-200, and what can you >>> tell >> me >>> about your experiences with your (purchased) nanoscope(s)? >>> >>> Sincerely, >>> >>> George >>> p.s. Please no need to clutter up the listserv with other people's >>> nanoscope references - I know how to use PubMed. For that matter, >>> I've replicated the results of the following two papers on my confocal's: >>> >>> Subdiffraction fluorescence imaging of biomolecular structure and >>> distributions with quantum dots. </pubmed/20600360> >>> >>> Heidbreder M, Endesfelder U, van de Linde S, Hennig S, Widera D, >>> Kaltschmidt B, Kaltschmidt C, Heilemann M. >>> >>> Biochim Biophys Acta. 2010 Oct;1803(10):1224-9. Epub 2010 Jun >>> 23.PMID: 20600360 >>> >>> >>> Quantum dot triexciton imaging with three-dimensional subdiffraction >>> resolution. </pubmed/19453186> >>> >>> Hennig S, van de Linde S, Heilemann M, Sauer M. >>> >>> Nano Lett. 2009 Jun;9(6):2466-70.PMID: 19453186 >>> >>> I have used <1 Airy unit pinhole to improve triexciton resolution >> further. >>> I have not tried doing 3D deconvolution on the data. >>> >>> >>> -- >>> >>> >>> George McNamara, PhD >>> Analytical Imaging Core Facility >>> University of Miami >>> >> ISTITUTO ITALIANO DI TECNOLOGIA Prof. Alberto Diaspro Scientific Head Nanophysics Via Morego, 30 16163 Genova Tel: +39-010.71.781.503 Fax +39-010-72.03.21 Mobile +39-3666719968 www.iit.it [hidden email] |
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** David, when it comes to the software side of things for STORM/PALM, you might consider Henriques Ricardo's QuickPALM ImageJ/MicroManager plugin: Henriques, R., et al., Quickpalm: 3d real-time photoactivation nanoscopy image processing in ImageJ. Nat Meth, 2010. 7(5): p. 339-340. Henriques has also put together a few youtube videos guiding a user step-by-step on how to put it all together (hardware and software): http://www.youtube.com/user/paxcal Cheers! John Oreopoulos Research Assistant Spectral Applied Research 9078 Leslie Street, Unit 11 Richmond Hill Ontario, Canada www.spectral.ca On 2011-01-31, at 10:05 AM, David Burk wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Since the topic of home built super-resolution systems has been brought up I was wondering if anyone had a very simple "Super Res for Dummies" type manual for building such a system. While I feel quite capable of following directions and assembling components together I have a certain degree of trepidation when it comes to figuring out how to drive all the components software-wise. I will admit I lack programming experience and limit myself to making ImageJ macros (poorly) or Cell Profiler pipelines - not writing code in Matlab to open shutters and such. I am aware that many articles describe their setups and give overview diagrams but, from a true schematic standpoint, they lack sufficient detail for someone as rigidly OCD as myself. Perhaps there is a class or course offered somewhere that covers this "roll your own" approach and I have yet to convince Google to divulge that information. > > Along the same lines, I am very interested in trying to construct an optical projection tomography system for our facility and, again, while I know many labs have built their own systems and published details of them, some critical details elude me. Have any listers built their own OPT rig and, if so, could you provide a detailed component list as well as assembly instructions? Personally, I would be more than willing to come visit a lab that has implemented their own solution to the FPALM/STORM and/or OPT method if they wouldn't mind spending a little time answering what they most likely would think are silly questions (and would allow me to take pictures of their system). > > David > > David H. Burk, PhD > Cell Biology and Bioimaging Core > Pennington Biomedical Research Center > Baton Rouge, LA 70808 > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Alberto Diaspro > Sent: Saturday, January 29, 2011 10:47 AM > To: [hidden email] > Subject: Re: Who has purchased a fluorescence nanoscope? > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > At the Italian Institute of Technology, we got a Leica STED-CW and home built an FPALM on an inverted Nikon thanks to software sharing from Sam Hess Lab, University of Maine. We home built a WLL and a CW STED controlled by MPI Nanobiophotonics software. We are currently interested n the Nikon N-STORM. > All the best > Alby > Il giorno 29/gen/2011, alle ore 17.19, Christophe Leterrier ha scritto: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> The Bordeaux Imaging Center in Bordeaux, France has a STED and a >> TIRF/PALM http://www.bic.u-bordeaux2.fr/ >> >> PALM/STORM setups are becoming more common (I'm aware of 2 setups >> already running in Marseille), because (at least this is what the >> people who did it told me) it is quite easy to add to an existing TIRF >> setup (provided you find software for the detection and localisation >> of individual fluorophores, but there is now an available ImageJ plugin for that). >> >> >> -- >> Christophe Leterrier >> Postdoc >> INSERM UMR641 // Ionic channels Lab >> IFR Jean Roche, Mediterranée University Marseille, France >> [hidden email] >> >> >> >> >> On Sat, Jan 29, 2011 at 16:52, G. Esteban Fernandez < >> [hidden email]> wrote: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> UCLA has a STED >>> >>> http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791 >>> <http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791> >>> http://www.cnsi.ucla.edu/staticpages/core-facilities#alms >>> >>> >>> <http://www.cnsi.ucla.edu/staticpages/core-facilities#alms> >>> >>> On Sat, Jan 29, 2011 at 7:32 AM, George McNamara >>> <[hidden email]>wrote: >>> >>>> ***** >>>> To join, leave or search the confocal microscopy listserv, go to: >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>>> ***** >>>> >>>> Dear Confocal listserv, >>>> >>>> Who has purchased a fluorescence nanoscope? What can you tell me >>>> about >>> your >>>> experiences - either here or privately? >>>> >>>> I see 11 OMX labs listed at http://www.api.com/omx-labs.asp (may be >>>> out >>> of >>>> date or some customers may be shy). >>>> >>>> I am aware of STED systems in the USA at Yale Univ, NIH and I was >>>> told >>> San >>>> Diego (have not found where), and possibly UC Denver. Paul French >>> apparently >>>> did his own upgrade of a Leica SP2 ( >>>> http://www.focusonmicroscopy.org/2008/PDF/070_Auksorius.pdf and a >>>> Leica Scientific Forum video ... see also related work at >>>> ftp://ftp.espci.fr/incoming/Gilles/INSERM-STED-x6.pdf ). >>>> >>>> I am also aware of one 4pi microscope in the USA. >>>> >>>> So, who has N-SIM, N-STORM, SR-SIM, PALm, SR-200, and what can you >>>> tell >>> me >>>> about your experiences with your (purchased) nanoscope(s)? >>>> >>>> Sincerely, >>>> >>>> George >>>> p.s. Please no need to clutter up the listserv with other people's >>>> nanoscope references - I know how to use PubMed. For that matter, >>>> I've replicated the results of the following two papers on my confocal's: >>>> >>>> Subdiffraction fluorescence imaging of biomolecular structure and >>>> distributions with quantum dots. </pubmed/20600360> >>>> >>>> Heidbreder M, Endesfelder U, van de Linde S, Hennig S, Widera D, >>>> Kaltschmidt B, Kaltschmidt C, Heilemann M. >>>> >>>> Biochim Biophys Acta. 2010 Oct;1803(10):1224-9. Epub 2010 Jun >>>> 23.PMID: 20600360 >>>> >>>> >>>> Quantum dot triexciton imaging with three-dimensional subdiffraction >>>> resolution. </pubmed/19453186> >>>> >>>> Hennig S, van de Linde S, Heilemann M, Sauer M. >>>> >>>> Nano Lett. 2009 Jun;9(6):2466-70.PMID: 19453186 >>>> >>>> I have used <1 Airy unit pinhole to improve triexciton resolution >>> further. >>>> I have not tried doing 3D deconvolution on the data. >>>> >>>> >>>> -- >>>> >>>> >>>> George McNamara, PhD >>>> Analytical Imaging Core Facility >>>> University of Miami >>>> >>> > > > > ISTITUTO ITALIANO > DI TECNOLOGIA > > Prof. Alberto Diaspro > Scientific Head > Nanophysics > Via Morego, 30 16163 Genova > Tel: +39-010.71.781.503 > Fax +39-010-72.03.21 > Mobile +39-3666719968 > www.iit.it > [hidden email] |
In reply to this post by David Burk
I've built STORM/PALM system and can confirm that it's not t
***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi David, I've built STORM/PALM system and can confirm that it's not too difficult. That said, every system will usually be quite different, depending on the specific requirements and abilities making a 'one size fits all' instruction manual very difficult to write. In general it'd be fair to say that some optics/laser design & alignment experience is desirable when it comes to getting the hardware side working, and it might be worth getting someone from the local physics department to help you. Programming experience is also a plus, although something like 'QuickPALM' (as mentioned by John) might give you a leg up (you'd probably still need to do a bit of tweaking to get everything working on your system). I'm personally not a huge fan of ImageJ/umanager/ Java in general, so would probably look for another solution, but this might be personal preference. We use custom, in house, software which I think is a lot more mature, but which is currently not particularly well documented. It also pays to give some thought to what variants of PALM/STORM you want to support as this has a huge influence on the setup. If your users are looking at fixed specimens with antibody labelling, something like dSTORM is probably the easiest to implement (single laser, no complicated switching patterns, can get away without software control of shutters etc ...). The two aspects of the hardware that you can't really get off the shelf components for and are probably going to need to play with are: - the laser coupling (commercial TIRF couplers typically give too little power over too large a field of view), - the focus mechanism (most commercial z-drives show quite a lot of drift - so you're going to either need some form of active stabilisation, or to redesign the focus mechanism - we ended up using a PIFoc piezo focusser on a custom metal bracket & doing away with the microscopes focus mechanism completely). If you've got any specific questions, you're welcome to get in touch, cheers, David ----- Original Message ---- From: David Burk <[hidden email]> To: [hidden email] Sent: Tue, 1 February, 2011 4:05:53 AM Subject: Building a STORM/FPALM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Since the topic of home built super-resolution systems has been brought up I was wondering if anyone had a very simple "Super Res for Dummies" type manual for building such a system. While I feel quite capable of following directions and assembling components together I have a certain degree of trepidation when it comes to figuring out how to drive all the components software-wise. I will admit I lack programming experience and limit myself to making ImageJ macros (poorly) or Cell Profiler pipelines - not writing code in Matlab to open shutters and such. I am aware that many articles describe their setups and give overview diagrams but, from a true schematic standpoint, they lack sufficient detail for someone as rigidly OCD as myself. Perhaps there is a class or course offered somewhere that covers this "roll your own" approach and I have yet to convince Google to divulge that information. Along the same lines, I am very interested in trying to construct an optical projection tomography system for our facility and, again, while I know many labs have built their own systems and published details of them, some critical details elude me. Have any listers built their own OPT rig and, if so, could you provide a detailed component list as well as assembly instructions? Personally, I would be more than willing to come visit a lab that has implemented their own solution to the FPALM/STORM and/or OPT method if they wouldn't mind spending a little time answering what they most likely would think are silly questions (and would allow me to take pictures of their system). David David H. Burk, PhD Cell Biology and Bioimaging Core Pennington Biomedical Research Center Baton Rouge, LA 70808 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Alberto Diaspro Sent: Saturday, January 29, 2011 10:47 AM To: [hidden email] Subject: Re: Who has purchased a fluorescence nanoscope? ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** At the Italian Institute of Technology, we got a Leica STED-CW and home built an FPALM on an inverted Nikon thanks to software sharing from Sam Hess Lab, University of Maine. We home built a WLL and a CW STED controlled by MPI Nanobiophotonics software. We are currently interested n the Nikon N-STORM. All the best Alby Il giorno 29/gen/2011, alle ore 17.19, Christophe Leterrier ha scritto: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > The Bordeaux Imaging Center in Bordeaux, France has a STED and a > TIRF/PALM http://www.bic.u-bordeaux2.fr/ > > PALM/STORM setups are becoming more common (I'm aware of 2 setups > already running in Marseille), because (at least this is what the > people who did it told me) it is quite easy to add to an existing TIRF > setup (provided you find software for the detection and localisation > of individual fluorophores, but there is now an available ImageJ plugin for >that). > > > -- > Christophe Leterrier > Postdoc > INSERM UMR641 // Ionic channels Lab > IFR Jean Roche, Mediterranée University Marseille, France > [hidden email] > > > > > On Sat, Jan 29, 2011 at 16:52, G. Esteban Fernandez < > [hidden email]> wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> UCLA has a STED >> >> http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791 >> <http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791> >> http://www.cnsi.ucla.edu/staticpages/core-facilities#alms >> >> >> <http://www.cnsi.ucla.edu/staticpages/core-facilities#alms> >> >> On Sat, Jan 29, 2011 at 7:32 AM, George McNamara >> <[hidden email]>wrote: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> Dear Confocal listserv, >>> >>> Who has purchased a fluorescence nanoscope? What can you tell me >>> about >> your >>> experiences - either here or privately? >>> >>> I see 11 OMX labs listed at http://www.api.com/omx-labs.asp (may be >>> out >> of >>> date or some customers may be shy). >>> >>> I am aware of STED systems in the USA at Yale Univ, NIH and I was >>> told >> San >>> Diego (have not found where), and possibly UC Denver. Paul French >> apparently >>> did his own upgrade of a Leica SP2 ( >>> http://www.focusonmicroscopy.org/2008/PDF/070_Auksorius.pdf and a >>> Leica Scientific Forum video ... see also related work at >>> ftp://ftp.espci.fr/incoming/Gilles/INSERM-STED-x6.pdf ). >>> >>> I am also aware of one 4pi microscope in the USA. >>> >>> So, who has N-SIM, N-STORM, SR-SIM, PALm, SR-200, and what can you >>> tell >> me >>> about your experiences with your (purchased) nanoscope(s)? >>> >>> Sincerely, >>> >>> George >>> p.s. Please no need to clutter up the listserv with other people's >>> nanoscope references - I know how to use PubMed. For that matter, >>> I've replicated the results of the following two papers on my confocal's: >>> >>> Subdiffraction fluorescence imaging of biomolecular structure and >>> distributions with quantum dots. </pubmed/20600360> >>> >>> Heidbreder M, Endesfelder U, van de Linde S, Hennig S, Widera D, >>> Kaltschmidt B, Kaltschmidt C, Heilemann M. >>> >>> Biochim Biophys Acta. 2010 Oct;1803(10):1224-9. Epub 2010 Jun >>> 23.PMID: 20600360 >>> >>> >>> Quantum dot triexciton imaging with three-dimensional subdiffraction >>> resolution. </pubmed/19453186> >>> >>> Hennig S, van de Linde S, Heilemann M, Sauer M. >>> >>> Nano Lett. 2009 Jun;9(6):2466-70.PMID: 19453186 >>> >>> I have used <1 Airy unit pinhole to improve triexciton resolution >> further. >>> I have not tried doing 3D deconvolution on the data. >>> >>> >>> -- >>> >>> >>> George McNamara, PhD >>> Analytical Imaging Core Facility >>> University of Miami >>> >> ISTITUTO ITALIANO DI TECNOLOGIA Prof. Alberto Diaspro Scientific Head Nanophysics Via Morego, 30 16163 Genova Tel: +39-010.71.781.503 Fax +39-010-72.03.21 Mobile +39-3666719968 www.iit.it [hidden email] |
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** For those of you who might be interested in just getting a quick overview of STORM/PALM as well as several other superresolution techniques, I've just posted the BioPhotonics article at our website, www.MicroscopyEducation.com. Just click on the "Library" and look for the listing. Best regards, Barbara Foster, President and Sr. Consultant Microscopy/Microscopy Education 7101 Royal Glen Trail, Suite A McKinney TX 75070 P: (972)924-5310 Skype: fostermme W: www.MicroscopyEducation.com At 05:59 PM 1/31/2011, you wrote: >I've built STORM/PALM system and can confirm that it's not t >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >Hi David, I've built STORM/PALM system and can >confirm that it's not too difficult. That said, >every system will usually be quite different, >depending on the specific requirements and >abilities making a 'one size fits all' >instruction manual very difficult to write. In >general it'd be fair to say that some >optics/laser design & alignment experience is >desirable when it comes to getting the hardware >side working, and it might be worth getting >someone from the local physics department to >help you. Programming experience is also a plus, >although something like 'QuickPALM' (as >mentioned by John) might give you a leg up >(you'd probably still need to do a bit of >tweaking to get everything working on your >system). I'm personally not a huge fan of >ImageJ/umanager/ Java in general, so would >probably look for another solution, but this >might be personal preference. We use custom, in >house, software which I think is a lot more >mature, but which is currently not particularly >well documented. It also pays to give some >thought to what variants of PALM/STORM you want >to support as this has a huge influence on the >setup. If your users are looking at fixed >specimens with antibody labelling, something >like dSTORM is probably the easiest to implement >(single laser, no complicated switching >patterns, can get away without software control >of shutters etc ...). The two aspects of the >hardware that you can't really get off the shelf >components for and are probably going to need to >play with are: - the laser coupling (commercial >TIRF couplers typically give too little power >over too large a field of view), - the focus >mechanism (most commercial z-drives show quite a >lot of drift - so you're going to either need >some form of active stabilisation, or to >redesign the focus mechanism - we ended up using >a PIFoc piezo focusser on a custom metal bracket >& doing away with the microscopes focus >mechanism completely). If you've got any >specific questions, you're welcome to get in >touch, cheers, David ----- Original Message ---- >From: David Burk <[hidden email]> To: >[hidden email] Sent: Tue, 1 >February, 2011 4:05:53 AM Subject: Building a >STORM/FPALM ***** To join, leave or search the >confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** Since the topic of home built >super-resolution systems has been brought up I >was wondering if anyone had a very simple "Super >Res for Dummies" type manual for building such a >system. While I feel quite capable of following >directions and assembling components together I >have a certain degree of trepidation when it >comes to figuring out how to drive all the >components software-wise. I will admit I lack >programming experience and limit myself to >making ImageJ macros (poorly) or Cell Profiler >pipelines - not writing code in Matlab to open >shutters and such. I am aware that many >articles describe their setups and give overview >diagrams but, from a true schematic standpoint, >they lack sufficient detail for someone as >rigidly OCD as myself. Perhaps there is a class >or course offered somewhere that covers this >"roll your own" approach and I have yet to >convince Google to divulge that >information. Along the same lines, I am very >interested in trying to construct an optical >projection tomography system for our facility >and, again, while I know many labs have built >their own systems and published details of them, >some critical details elude me. Have any >listers built their own OPT rig and, if so, >could you provide a detailed component list as >well as assembly instructions? Personally, I >would be more than willing to come visit a lab >that has implemented their own solution to the >FPALM/STORM and/or OPT method if they wouldn't >mind spending a little time answering what they >most likely would think are silly questions (and >would allow me to take pictures of their >system). David David H. Burk, PhD Cell Biology >and Bioimaging Core Pennington Biomedical >Research Center Baton Rouge, LA 70808 >-----Original Message----- From: Confocal >Microscopy List >[mailto:[hidden email]] On >Behalf Of Alberto Diaspro Sent: Saturday, >January 29, 2011 10:47 AM To: >[hidden email] Subject: Re: >Who has purchased a fluorescence nanoscope? >***** To join, leave or search the confocal >microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** At the Italian Institute of Technology, we >got a Leica STED-CW and home built an FPALM on >an inverted Nikon thanks to software sharing >from Sam Hess Lab, University of Maine. We home >built a WLL and a CW STED controlled by MPI >Nanobiophotonics software. We are currently >interested n the Nikon N-STORM. All the best >Alby Il giorno 29/gen/2011, alle ore 17.19, >Christophe Leterrier ha scritto: > ***** > To >join, leave or search the confocal microscopy >listserv, go to: > >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > The Bordeaux Imaging Center in >Bordeaux, France has a STED and a > TIRF/PALM >http://www.bic.u-bordeaux2.fr/ > > PALM/STORM >setups are becoming more common (I'm aware of 2 >setups > already running in Marseille), because >(at least this is what the > people who did it >told me) it is quite easy to add to an existing >TIRF > setup (provided you find software for the >detection and localisation > of individual >fluorophores, but there is now an available >ImageJ plugin for >that). > > > -- > Christophe >Leterrier > Postdoc > INSERM UMR641 // Ionic >channels Lab > IFR Jean Roche, Mediterranée >University Marseille, France > >[hidden email] > > > > > On >Sat, Jan 29, 2011 at 16:52, G. Esteban Fernandez >< > [hidden email]> wrote: > >> >***** >> To join, leave or search the confocal >microscopy listserv, go to: >> >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >> ***** >> >> UCLA has a STED >> >> >http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791 > >> ><http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791> > >> >http://www.cnsi.ucla.edu/staticpages/core-facilities#alms > >> >> >> ><http://www.cnsi.ucla.edu/staticpages/core-facilities#alms> > >> >> On Sat, Jan 29, 2011 at 7:32 AM, George >McNamara >> ><[hidden email]>wrote: >> >>> >***** >>> To join, leave or search the confocal >microscopy listserv, go to: >>> >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > >>> ***** >>> >>> Dear Confocal >listserv, >>> >>> Who has purchased a >fluorescence nanoscope? What can you tell me >>> >about >> your >>> experiences - either here or >privately? >>> >>> I see 11 OMX labs listed at >http://www.api.com/omx-labs.asp (may be >>> >out >> of >>> date or some customers may be >shy). >>> >>> I am aware of STED systems in the >USA at Yale Univ, NIH and I was >>> told >> >San >>> Diego (have not found where), and >possibly UC Denver. Paul French >> >apparently >>> did his own upgrade of a Leica >SP2 ( >>> >http://www.focusonmicroscopy.org/2008/PDF/070_Auksorius.pdf >and a >>> Leica Scientific Forum video ... see >also related work at >>> >ftp://ftp.espci.fr/incoming/Gilles/INSERM-STED-x6.pdf >). >>> >>> I am also aware of one 4pi microscope >in the USA. >>> >>> So, who has N-SIM, N-STORM, >SR-SIM, PALm, SR-200, and what can you >>> >tell >> me >>> about your experiences with your >(purchased) nanoscope(s)? >>> >>> >Sincerely, >>> >>> George >>> p.s. Please no >need to clutter up the listserv with other >people's >>> nanoscope references - I know how >to use PubMed. For that matter, >>> I've >replicated the results of the following two >papers on my confocal's: >>> >>> Subdiffraction >fluorescence imaging of biomolecular structure >and >>> distributions with quantum dots. ></pubmed/20600360> >>> >>> Heidbreder M, >Endesfelder U, van de Linde S, Hennig S, Widera >D, >>> Kaltschmidt B, Kaltschmidt C, Heilemann >M. >>> >>> Biochim Biophys Acta. 2010 >Oct;1803(10):1224-9. Epub 2010 Jun >>> 23.PMID: >20600360 >>> >>> >>> Quantum dot triexciton >imaging with three-dimensional >subdiffraction >>> resolution. ></pubmed/19453186> >>> >>> Hennig S, van de >Linde S, Heilemann M, Sauer M. >>> >>> Nano >Lett. 2009 Jun;9(6):2466-70.PMID: >19453186 >>> >>> I have used <1 Airy unit >pinhole to improve triexciton resolution >> >further. >>> I have not tried doing 3D >deconvolution on the data. >>> >>> >>> >-- >>> >>> >>> George McNamara, PhD >>> >Analytical Imaging Core Facility >>> University >of Miami >>> >> ISTITUTO ITALIANO DI TECNOLOGIA >Prof. Alberto Diaspro Scientific Head >Nanophysics Via Morego, 30 16163 Genova Tel: >+39-010.71.781.503 Fax +39-010-72.03.21 Mobile >+39-3666719968 www.iit.it [hidden email] |
In reply to this post by John Oreopoulos
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Just to make it clear, his name is Ricardo Henriques. He showed me an early version of his software and I was quite amazed by its speed. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net ________________________________ From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of John Oreopoulos Sent: Tuesday, 1 February 2011 3:19 AM To: [hidden email] Subject: Re: Building a STORM/FPALM ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** David, when it comes to the software side of things for STORM/PALM, you might consider Henriques Ricardo's QuickPALM ImageJ/MicroManager plugin: Henriques, R., et al., Quickpalm: 3d real-time photoactivation nanoscopy image processing in ImageJ. Nat Meth, 2010. 7(5): p. 339-340. Henriques has also put together a few youtube videos guiding a user step-by-step on how to put it all together (hardware and software): http://www.youtube.com/user/paxcal Cheers! John Oreopoulos Research Assistant Spectral Applied Research 9078 Leslie Street, Unit 11 Richmond Hill Ontario, Canada www.spectral.ca On 2011-01-31, at 10:05 AM, David Burk wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Since the topic of home built super-resolution systems has been brought up I was wondering if anyone had a very simple "Super Res for Dummies" type manual for building such a system. While I feel quite capable of following directions and assembling components together I have a certain degree of trepidation when it comes to figuring out how to drive all the components software-wise. I will admit I lack programming experience and limit myself to making ImageJ macros (poorly) or Cell Profiler pipelines - not writing code in Matlab to open shutters and such. I am aware that many articles describe their setups and give overview diagrams but, from a true schematic standpoint, they lack sufficient detail for someone as rigidly OCD as myself. Perhaps there is a class or course offered somewhere that covers this "roll your own" approach and I have yet to convince Google to divulge that information. > > Along the same lines, I am very interested in trying to construct an optical projection tomography system for our facility and, again, while I know many labs have built their own systems and published details of them, some critical details elude me. Have any listers built their own OPT rig and, if so, could you provide a detailed component list as well as assembly instructions? Personally, I would be more than willing to come visit a lab that has implemented their own solution to the FPALM/STORM and/or OPT method if they wouldn't mind spending a little time answering what they most likely would think are silly questions (and would allow me to take pictures of their system). > > David > > David H. Burk, PhD > Cell Biology and Bioimaging Core > Pennington Biomedical Research Center > Baton Rouge, LA 70808 > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Alberto Diaspro > Sent: Saturday, January 29, 2011 10:47 AM > To: [hidden email] > Subject: Re: Who has purchased a fluorescence nanoscope? > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > At the Italian Institute of Technology, we got a Leica STED-CW and home built an FPALM on an inverted Nikon thanks to software sharing from Sam Hess Lab, University of Maine. We home built a WLL and a CW STED controlled by MPI Nanobiophotonics software. We are currently interested n the Nikon N-STORM. > All the best > Alby > Il giorno 29/gen/2011, alle ore 17.19, Christophe Leterrier ha scritto: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> The Bordeaux Imaging Center in Bordeaux, France has a STED and a >> TIRF/PALM http://www.bic.u-bordeaux2.fr/ >> >> PALM/STORM setups are becoming more common (I'm aware of 2 setups >> already running in Marseille), because (at least this is what the >> people who did it told me) it is quite easy to add to an existing TIRF >> setup (provided you find software for the detection and localisation >> of individual fluorophores, but there is now an available ImageJ plugin for that). >> >> >> -- >> Christophe Leterrier >> Postdoc >> INSERM UMR641 // Ionic channels Lab >> IFR Jean Roche, Mediterranée University Marseille, France >> [hidden email] >> >> >> >> >> On Sat, Jan 29, 2011 at 16:52, G. Esteban Fernandez < >> [hidden email]> wrote: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> UCLA has a STED >>> >>> http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791 >>> <http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791> >>> http://www.cnsi.ucla.edu/staticpages/core-facilities#alms >>> >>> >>> <http://www.cnsi.ucla.edu/staticpages/core-facilities#alms> >>> >>> On Sat, Jan 29, 2011 at 7:32 AM, George McNamara >>> <[hidden email]>wrote: >>> >>>> ***** >>>> To join, leave or search the confocal microscopy listserv, go to: >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>>> ***** >>>> >>>> Dear Confocal listserv, >>>> >>>> Who has purchased a fluorescence nanoscope? What can you tell me >>>> about >>> your >>>> experiences - either here or privately? >>>> >>>> I see 11 OMX labs listed at http://www.api.com/omx-labs.asp (may be >>>> out >>> of >>>> date or some customers may be shy). >>>> >>>> I am aware of STED systems in the USA at Yale Univ, NIH and I was >>>> told >>> San >>>> Diego (have not found where), and possibly UC Denver. Paul French >>> apparently >>>> did his own upgrade of a Leica SP2 ( >>>> http://www.focusonmicroscopy.org/2008/PDF/070_Auksorius.pdf and a >>>> Leica Scientific Forum video ... see also related work at >>>> ftp://ftp.espci.fr/incoming/Gilles/INSERM-STED-x6.pdf ). >>>> >>>> I am also aware of one 4pi microscope in the USA. >>>> >>>> So, who has N-SIM, N-STORM, SR-SIM, PALm, SR-200, and what can you >>>> tell >>> me >>>> about your experiences with your (purchased) nanoscope(s)? >>>> >>>> Sincerely, >>>> >>>> George >>>> p.s. Please no need to clutter up the listserv with other people's >>>> nanoscope references - I know how to use PubMed. For that matter, >>>> I've replicated the results of the following two papers on my confocal's: >>>> >>>> Subdiffraction fluorescence imaging of biomolecular structure and >>>> distributions with quantum dots. </pubmed/20600360> >>>> >>>> Heidbreder M, Endesfelder U, van de Linde S, Hennig S, Widera D, >>>> Kaltschmidt B, Kaltschmidt C, Heilemann M. >>>> >>>> Biochim Biophys Acta. 2010 Oct;1803(10):1224-9. Epub 2010 Jun >>>> 23.PMID: 20600360 >>>> >>>> >>>> Quantum dot triexciton imaging with three-dimensional subdiffraction >>>> resolution. </pubmed/19453186> >>>> >>>> Hennig S, van de Linde S, Heilemann M, Sauer M. >>>> >>>> Nano Lett. 2009 Jun;9(6):2466-70.PMID: 19453186 >>>> >>>> I have used <1 Airy unit pinhole to improve triexciton resolution >>> further. >>>> I have not tried doing 3D deconvolution on the data. >>>> >>>> >>>> -- >>>> >>>> >>>> George McNamara, PhD >>>> Analytical Imaging Core Facility >>>> University of Miami >>>> >>> > > > > ISTITUTO ITALIANO > DI TECNOLOGIA > > Prof. Alberto Diaspro > Scientific Head > Nanophysics > Via Morego, 30 16163 Genova > Tel: +39-010.71.781.503 > Fax +39-010-72.03.21 > Mobile +39-3666719968 > www.iit.it > [hidden email] ________________________________ No virus found in this message. Checked by AVG - www.avg.com Version: 10.0.1204 / Virus Database: 1435/3413 - Release Date: 01/30/11 |
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Whoops, sorry about that. John Oreopoulos On 2011-02-01, at 2:26 AM, Guy Cox <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Just to make it clear, his name is Ricardo Henriques. He showed me an early version of his software and I was quite amazed by its speed. > > > > Guy > > > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) > Electron Microscope Unit, Madsen Building F09, > University of Sydney, NSW 2006 > ______________________________________________ > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > > > ________________________________ > > From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of John Oreopoulos > Sent: Tuesday, 1 February 2011 3:19 AM > To: [hidden email] > Subject: Re: Building a STORM/FPALM > > > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > David, when it comes to the software side of things for STORM/PALM, you might consider Henriques Ricardo's QuickPALM ImageJ/MicroManager plugin: > > Henriques, R., et al., Quickpalm: 3d real-time photoactivation nanoscopy image processing in ImageJ. Nat Meth, 2010. 7(5): p. 339-340. > > Henriques has also put together a few youtube videos guiding a user step-by-step on how to put it all together (hardware and software): > > http://www.youtube.com/user/paxcal > > Cheers! > > John Oreopoulos > Research Assistant > Spectral Applied Research > 9078 Leslie Street, Unit 11 > Richmond Hill > Ontario, Canada > www.spectral.ca > > > On 2011-01-31, at 10:05 AM, David Burk wrote: > >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> Since the topic of home built super-resolution systems has been brought up I was wondering if anyone had a very simple "Super Res for Dummies" type manual for building such a system. While I feel quite capable of following directions and assembling components together I have a certain degree of trepidation when it comes to figuring out how to drive all the components software-wise. I will admit I lack programming experience and limit myself to making ImageJ macros (poorly) or Cell Profiler pipelines - not writing code in Matlab to open shutters and such. I am aware that many articles describe their setups and give overview diagrams but, from a true schematic standpoint, they lack sufficient detail for someone as rigidly OCD as myself. Perhaps there is a class or course offered somewhere that covers this "roll your own" approach and I have yet to convince Google to divulge that information. >> >> Along the same lines, I am very interested in trying to construct an optical projection tomography system for our facility and, again, while I know many labs have built their own systems and published details of them, some critical details elude me. Have any listers built their own OPT rig and, if so, could you provide a detailed component list as well as assembly instructions? Personally, I would be more than willing to come visit a lab that has implemented their own solution to the FPALM/STORM and/or OPT method if they wouldn't mind spending a little time answering what they most likely would think are silly questions (and would allow me to take pictures of their system). >> >> David >> >> David H. Burk, PhD >> Cell Biology and Bioimaging Core >> Pennington Biomedical Research Center >> Baton Rouge, LA 70808 >> >> >> -----Original Message----- >> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Alberto Diaspro >> Sent: Saturday, January 29, 2011 10:47 AM >> To: [hidden email] >> Subject: Re: Who has purchased a fluorescence nanoscope? >> >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> At the Italian Institute of Technology, we got a Leica STED-CW and home built an FPALM on an inverted Nikon thanks to software sharing from Sam Hess Lab, University of Maine. We home built a WLL and a CW STED controlled by MPI Nanobiophotonics software. We are currently interested n the Nikon N-STORM. >> All the best >> Alby >> Il giorno 29/gen/2011, alle ore 17.19, Christophe Leterrier ha scritto: >> >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> The Bordeaux Imaging Center in Bordeaux, France has a STED and a >>> TIRF/PALM http://www.bic.u-bordeaux2.fr/ >>> >>> PALM/STORM setups are becoming more common (I'm aware of 2 setups >>> already running in Marseille), because (at least this is what the >>> people who did it told me) it is quite easy to add to an existing TIRF >>> setup (provided you find software for the detection and localisation >>> of individual fluorophores, but there is now an available ImageJ plugin for that). >>> >>> >>> -- >>> Christophe Leterrier >>> Postdoc >>> INSERM UMR641 // Ionic channels Lab >>> IFR Jean Roche, Mediterranée University Marseille, France >>> [hidden email] >>> >>> >>> >>> >>> On Sat, Jan 29, 2011 at 16:52, G. Esteban Fernandez < >>> [hidden email]> wrote: >>> >>>> ***** >>>> To join, leave or search the confocal microscopy listserv, go to: >>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>>> ***** >>>> >>>> UCLA has a STED >>>> >>>> http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791 >>>> <http://clms.cnsi.ucla.edu/cnsi/clms/equipment-list?search_lab=6791> >>>> http://www.cnsi.ucla.edu/staticpages/core-facilities#alms >>>> >>>> >>>> <http://www.cnsi.ucla.edu/staticpages/core-facilities#alms> >>>> >>>> On Sat, Jan 29, 2011 at 7:32 AM, George McNamara >>>> <[hidden email]>wrote: >>>> >>>>> ***** >>>>> To join, leave or search the confocal microscopy listserv, go to: >>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>>>> ***** >>>>> >>>>> Dear Confocal listserv, >>>>> >>>>> Who has purchased a fluorescence nanoscope? What can you tell me >>>>> about >>>> your >>>>> experiences - either here or privately? >>>>> >>>>> I see 11 OMX labs listed at http://www.api.com/omx-labs.asp (may be >>>>> out >>>> of >>>>> date or some customers may be shy). >>>>> >>>>> I am aware of STED systems in the USA at Yale Univ, NIH and I was >>>>> told >>>> San >>>>> Diego (have not found where), and possibly UC Denver. Paul French >>>> apparently >>>>> did his own upgrade of a Leica SP2 ( >>>>> http://www.focusonmicroscopy.org/2008/PDF/070_Auksorius.pdf and a >>>>> Leica Scientific Forum video ... see also related work at >>>>> ftp://ftp.espci.fr/incoming/Gilles/INSERM-STED-x6.pdf ). >>>>> >>>>> I am also aware of one 4pi microscope in the USA. >>>>> >>>>> So, who has N-SIM, N-STORM, SR-SIM, PALm, SR-200, and what can you >>>>> tell >>>> me >>>>> about your experiences with your (purchased) nanoscope(s)? >>>>> >>>>> Sincerely, >>>>> >>>>> George >>>>> p.s. Please no need to clutter up the listserv with other people's >>>>> nanoscope references - I know how to use PubMed. For that matter, >>>>> I've replicated the results of the following two papers on my confocal's: >>>>> >>>>> Subdiffraction fluorescence imaging of biomolecular structure and >>>>> distributions with quantum dots. </pubmed/20600360> >>>>> >>>>> Heidbreder M, Endesfelder U, van de Linde S, Hennig S, Widera D, >>>>> Kaltschmidt B, Kaltschmidt C, Heilemann M. >>>>> >>>>> Biochim Biophys Acta. 2010 Oct;1803(10):1224-9. Epub 2010 Jun >>>>> 23.PMID: 20600360 >>>>> >>>>> >>>>> Quantum dot triexciton imaging with three-dimensional subdiffraction >>>>> resolution. </pubmed/19453186> >>>>> >>>>> Hennig S, van de Linde S, Heilemann M, Sauer M. >>>>> >>>>> Nano Lett. 2009 Jun;9(6):2466-70.PMID: 19453186 >>>>> >>>>> I have used <1 Airy unit pinhole to improve triexciton resolution >>>> further. >>>>> I have not tried doing 3D deconvolution on the data. >>>>> >>>>> >>>>> -- >>>>> >>>>> >>>>> George McNamara, PhD >>>>> Analytical Imaging Core Facility >>>>> University of Miami >>>>> >>>> >> >> >> >> ISTITUTO ITALIANO >> DI TECNOLOGIA >> >> Prof. Alberto Diaspro >> Scientific Head >> Nanophysics >> Via Morego, 30 16163 Genova >> Tel: +39-010.71.781.503 >> Fax +39-010-72.03.21 >> Mobile +39-3666719968 >> www.iit.it >> [hidden email] > > ________________________________ > > No virus found in this message. > Checked by AVG - www.avg.com > Version: 10.0.1204 / Virus Database: 1435/3413 - Release Date: 01/30/11 |
In reply to this post by David Burk
I have two problems, both in Arabidopsis.
First, has an ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I have two problems, both in Arabidopsis. First, has anyone on this list done FRET in live plant cells? If so, may I contact you off list for a few pointers? The entire cytoplasmic streaming and movement of organelles bit has me a bit stumped. Secondly, we have GFP which is targeting two organelles. The question is if the amounts going to each organelle can be quantified. The issue being one is roughly 10x as bright. I am unsure if any of the various settings on a Nikon A1 are linear, as it is impossible to image the two organelles in the same cell and have them be on the same intensity scale. Any pointers or ideas are greatly appreciated. Christian |
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