30x30x1 nm, 7 colors
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** 30x30x1 nm, 7 colors ... see figure 1 for colors - the 7 'colors' can probably be extended to more for a bit more money. http://www.nature.com/nature/journal/v481/n7382/extref/nature10734-s1.pdf The same lab has the adjacent article - http://www.nature.com/nature/journal/v481/n7382/extref/nature10745-s1.pdf 30x30x1 nm (yes, nanometer) is very nice - perhaps the fluorescence nanoscope developers should spend a bit less time on pushing from the current ~20x20x50 nm (which should get to 10x10x25 nm by a certain improvement in light output while on) and spend a bit more time on producing making new biomedical discoveries. I may say a few words about this approach during my short talk at the ABRF nanoscopy session in Orlando in late March. I might start the talk with the nice "SnapShot; light Microscopy" in Cell (Nov 23, 2011 issue - I was able to download from home) http://download.cell.com/pdf/PIIS0092867411013535.pdf?intermediate=true |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** This MIMS seems like a cool technique, if your question can be solved by investigating the surface of a dead sample (at least I assume the sample is dead, if not before then after the ion bombardement; certainly if the cell surface was sputtered as for Steinhauser et al. Fig 1a). They say the maximum field size is 80 x 80 µm (Methods to Steinhauser et al.) With an xy-resolution of 28 nm (Supp Fig 1 legend) according to the Nyquist criterion that would be >6500 px squared, probably too many. But let us assume a standard 512x512 px frame for some appropriate ROI, has anybody an idea what the frame time is? Steffen On 07.02.2012 04:14, George McNamara wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > 30x30x1 nm, 7 colors ... see figure 1 for colors - the 7 'colors' can > probably be extended to more for a bit more money. > http://www.nature.com/nature/journal/v481/n7382/extref/nature10734-s1.pdf > The same lab has the adjacent article - > http://www.nature.com/nature/journal/v481/n7382/extref/nature10745-s1.pdf > > 30x30x1 nm (yes, nanometer) is very nice - perhaps the fluorescence > nanoscope developers should spend a bit less time on pushing from the > current ~20x20x50 nm (which should get to 10x10x25 nm by a certain > improvement in light output while on) and spend a bit more time on > producing making new biomedical discoveries. > > I may say a few words about this approach during my short talk at the > ABRF nanoscopy session in Orlando in late March. > I might start the talk with the nice "SnapShot; light Microscopy" in > Cell (Nov 23, 2011 issue - I was able to download from home) > > http://download.cell.com/pdf/PIIS0092867411013535.pdf?intermediate=true > -- ------------------------------------------------------------ Steffen Dietzel, PD Dr. rer. nat Ludwig-Maximilians-Universität München Walter-Brendel-Zentrum für experimentelle Medizin (WBex) Head of light microscopy Mail room: Marchioninistr. 15, D-81377 München Building location: Marchioninistr. 27, München-Großhadern |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I guess the sample is not only dead (in vacuum?) but also destroyed inthe imaging process. The only time we used a focussed ion beam was to drill small holes in metal with better than optical precision. A cool technique, but for my taste the paper could have some more information about how the sample is prepared and comparisson to other high resolution techhniques. What is an imersion objective in the cotext of ion beam microscopy? best wishes Andreas -----Original Message----- From: Steffen Dietzel <[hidden email]> To: CONFOCALMICROSCOPY <[hidden email]> Sent: Thu, 9 Feb 2012 12:46 Subject: Re: 30x30x1 nm, 7 colors ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** This MIMS seems like a cool technique, if your question can be solved by investigating the surface of a dead sample (at least I assume the sample is dead, if not before then after the ion bombardement; certainly if the cell surface was sputtered as for Steinhauser et al. Fig 1a). They say the maximum field size is 80 x 80 µm (Methods to Steinhauser et al.) With an xy-resolution of 28 nm (Supp Fig 1 legend) according to the Nyquist criterion that would be >6500 px squared, probably too many. But let us assume a standard 512x512 px frame for some appropriate ROI, has anybody an idea what the frame time is? Steffen On 07.02.2012 04:14, George McNamara wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > 30x30x1 nm, 7 colors ... see figure 1 for colors - the 7 'colors' can > probably be extended to more for a bit more money. > http://www.nature.com/nature/journal/v481/n7382/extref/nature10734-s1.pdf > The same lab has the adjacent article - > http://www.nature.com/nature/journal/v481/n7382/extref/nature10745-s1.pdf > > 30x30x1 nm (yes, nanometer) is very nice - perhaps the fluorescence > nanoscope developers should spend a bit less time on pushing from the > current ~20x20x50 nm (which should get to 10x10x25 nm by a certain > improvement in light output while on) and spend a bit more time on > producing making new biomedical discoveries. > > I may say a few words about this approach during my short talk at the > ABRF nanoscopy session in Orlando in late March. > I might start the talk with the nice "SnapShot; light Microscopy" in > Cell (Nov 23, 2011 issue - I was able to download from home) > > http://download.cell.com/pdf/PIIS0092867411013535.pdf?intermediate=true > -- ------------------------------------------------------------ Steffen Dietzel, PD Dr. rer. nat Ludwig-Maximilians-Universität München Walter-Brendel-Zentrum für experimentelle Medizin (WBex) Head of light microscopy Mail room: Marchioninistr. 15, D-81377 München Building location: Marchioninistr. 27, München-Großhadern |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** More details in the research article + 2 commentaries in http://jbiol.com/content/5/October/2006 Volume 5 (2006) - October 2006 </content/5#5> next > </content/5/November/2006> up </content/5#5> < previous </content/5/August/2006> Research article ** * High-resolution quantitative imaging of mammalian and bacterial cells using stable isotope mass spectrometry <http://jbiol.com/content/5/6/20>* Claude Lechene, Francois Hillion, Greg McMahon, Douglas Benson, Alan M Kleinfeld, J Patrick Kampf, Daniel Distel, Yvette Luyten, Joseph Bonventre, Dirk Hentschel, Kwon Park, Susumu Ito, Martin Schwartz, Gilles Benichou, Georges Slodzian /Journal of Biology/ 2006, *5*:20 (5 October 2006) Abstract <http://jbiol.com/content/5/6/20/abstract> | Full text <http://jbiol.com/content/5/6/20> | PDF <http://jbiol.com/content/pdf/jbiol42.pdf> | PubMed <http://jbiol.com/pubmed/17010211> | Cited on BioMed Central <http://jbiol.com/content/5/6/20/about/citations-biomedcentral> | F1000 Biology <http://www.f1000biology.com/article/17010211>|/ /Editor’s summary <#> For the first time it is possible to image and quantify at nanometer resolution biological samples labeled with stable isotopes, using multi-isotope imaging mass spectrometry, applicable to all fields of biomedical research. Minireview ** * Biological imaging using secondary ions <http://jbiol.com/content/5/6/18>* Peter Williams /Journal of Biology/ 2006, *5*:18 (5 October 2006) Abstract <http://jbiol.com/content/5/6/18/abstract> | Full text <http://jbiol.com/content/5/6/18> | PDF <http://jbiol.com/content/pdf/jbiol49.pdf> | PubMed <http://jbiol.com/pubmed/17029649> Research news ** * Imaging with isotopes: high resolution and quantitation <http://jbiol.com/content/5/6/17>* Jonathan B Weitzman /Journal of Biology/ 2006, *5*:17 (5 October 2006) Full text <http://jbiol.com/content/5/6/17> | PDF <http://jbiol.com/content/pdf/jbiol48.pdf> On 2/9/2012 9:12 AM, Andreas Bruckbauer wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > I guess the sample is not only dead (in vacuum?) but also destroyed inthe imaging process. The only time we used a focussed ion beam was to drill small holes in metal with better than optical precision. A cool technique, but for my taste the paper could have some more information about how the sample is prepared and comparisson to other high resolution techhniques. What is an imersion objective in the cotext of ion beam microscopy? > > best wishes > > Andreas > > > > > > > > -----Original Message----- > From: Steffen Dietzel<[hidden email]> > To: CONFOCALMICROSCOPY<[hidden email]> > Sent: Thu, 9 Feb 2012 12:46 > Subject: Re: 30x30x1 nm, 7 colors > > > ***** > > To join, leave or search the confocal microscopy listserv, go to: > > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > > ***** > > > > This MIMS seems like a cool technique, if your question can be solved by > > investigating the surface of a dead sample (at least I assume the sample > > is dead, if not before then after the ion bombardement; certainly if the > > cell surface was sputtered as for Steinhauser et al. Fig 1a). > > > > They say the maximum field size is 80 x 80 µm (Methods to Steinhauser et > > al.) With an xy-resolution of 28 nm (Supp Fig 1 legend) according to the > > Nyquist criterion that would be>6500 px squared, probably too many. But > > let us assume a standard 512x512 px frame for some appropriate ROI, has > > anybody an idea what the frame time is? > > > > Steffen > > > > On 07.02.2012 04:14, George McNamara wrote: > > >> ***** >> > >> To join, leave or search the confocal microscopy listserv, go to: >> > >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> > >> ***** >> > >> > >> 30x30x1 nm, 7 colors ... see figure 1 for colors - the 7 'colors' can >> > >> probably be extended to more for a bit more money. >> > >> http://www.nature.com/nature/journal/v481/n7382/extref/nature10734-s1.pdf >> > >> The same lab has the adjacent article - >> > >> http://www.nature.com/nature/journal/v481/n7382/extref/nature10745-s1.pdf >> > >> > >> 30x30x1 nm (yes, nanometer) is very nice - perhaps the fluorescence >> > >> nanoscope developers should spend a bit less time on pushing from the >> > >> current ~20x20x50 nm (which should get to 10x10x25 nm by a certain >> > >> improvement in light output while on) and spend a bit more time on >> > >> producing making new biomedical discoveries. >> > >> > >> I may say a few words about this approach during my short talk at the >> > >> ABRF nanoscopy session in Orlando in late March. >> > >> I might start the talk with the nice "SnapShot; light Microscopy" in >> > >> Cell (Nov 23, 2011 issue - I was able to download from home) >> > >> > >> http://download.cell.com/pdf/PIIS0092867411013535.pdf?intermediate=true >> > >> > > > > > -- George McNamara, PhD Analytical Imaging Core Facility University of Miami |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** By 'sputter' they mean eject secondary ions (primary ion is Cs or O). To image deeper, keep imaging. They also mention a way to ablate layers of atoms faster. Pathologists have been using dead H&E tissue sections for clinical diagnosis for a century. Almost all TEM has been on dead specimens for over 50 years. For that matter, most fluorescence microscopy - and especially confocal - microscopy specimens for the past 25 years have been dead on arrival on the microscope. Dead is dead - doesn't matter how fast if it gets you the atomic (ions) info unavailable at the resolution needed to answer the question. More interesting is where to store, archive, process and analyze all this data. On 2/9/2012 7:44 AM, Steffen Dietzel wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > This MIMS seems like a cool technique, if your question can be solved > by investigating the surface of a dead sample (at least I assume the > sample is dead, if not before then after the ion bombardement; > certainly if the cell surface was sputtered as for Steinhauser et al. > Fig 1a). > > They say the maximum field size is 80 x 80 µm (Methods to Steinhauser > et al.) With an xy-resolution of 28 nm (Supp Fig 1 legend) according > to the Nyquist criterion that would be >6500 px squared, probably too > many. But let us assume a standard 512x512 px frame for some > appropriate ROI, has anybody an idea what the frame time is? > > Steffen > > On 07.02.2012 04:14, George McNamara wrote: >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> 30x30x1 nm, 7 colors ... see figure 1 for colors - the 7 'colors' can >> probably be extended to more for a bit more money. >> http://www.nature.com/nature/journal/v481/n7382/extref/nature10734-s1.pdf >> >> The same lab has the adjacent article - >> http://www.nature.com/nature/journal/v481/n7382/extref/nature10745-s1.pdf >> >> >> 30x30x1 nm (yes, nanometer) is very nice - perhaps the fluorescence >> nanoscope developers should spend a bit less time on pushing from the >> current ~20x20x50 nm (which should get to 10x10x25 nm by a certain >> improvement in light output while on) and spend a bit more time on >> producing making new biomedical discoveries. >> >> I may say a few words about this approach during my short talk at the >> ABRF nanoscopy session in Orlando in late March. >> I might start the talk with the nice "SnapShot; light Microscopy" in >> Cell (Nov 23, 2011 issue - I was able to download from home) >> >> http://download.cell.com/pdf/PIIS0092867411013535.pdf?intermediate=true >> > > -- George McNamara, PhD Analytical Imaging Core Facility University of Miami |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** George, I didn't mean to imply that dead is useless. I use dead samples whenever I can get away with it. Much less trouble than live cell or intravital microscopy, so I totally agree with you on that point. I merely mentioned this to suggest that super resolution light microscopy still has some advantages (potential advantages at least, I admit that sometimes I wonder how alive a sample still is after catching all the photons it requires for these techniques.) I disagree on the issue of speed however. If you need n=30, it will make an important practical difference whether your frame time is a minute or an hour. Thank you for the additional links in your other e-mail. Pretty fascinating. Steffen On 10.02.2012 02:04, George McNamara wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > By 'sputter' they mean eject secondary ions (primary ion is Cs or O). To > image deeper, keep imaging. They also mention a way to ablate layers of > atoms faster. > > Pathologists have been using dead H&E tissue sections for clinical > diagnosis for a century. Almost all TEM has been on dead specimens for > over 50 years. For that matter, most fluorescence microscopy - and > especially confocal - microscopy specimens for the past 25 years have > been dead on arrival on the microscope. > > Dead is dead - doesn't matter how fast if it gets you the atomic (ions) > info unavailable at the resolution needed to answer the question. More > interesting is where to store, archive, process and analyze all this data. > > > > > On 2/9/2012 7:44 AM, Steffen Dietzel wrote: >> ***** >> To join, leave or search the confocal microscopy listserv, go to: >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >> ***** >> >> This MIMS seems like a cool technique, if your question can be solved >> by investigating the surface of a dead sample (at least I assume the >> sample is dead, if not before then after the ion bombardement; >> certainly if the cell surface was sputtered as for Steinhauser et al. >> Fig 1a). >> >> They say the maximum field size is 80 x 80 µm (Methods to Steinhauser >> et al.) With an xy-resolution of 28 nm (Supp Fig 1 legend) according >> to the Nyquist criterion that would be >6500 px squared, probably too >> many. But let us assume a standard 512x512 px frame for some >> appropriate ROI, has anybody an idea what the frame time is? >> >> Steffen >> >> On 07.02.2012 04:14, George McNamara wrote: >>> ***** >>> To join, leave or search the confocal microscopy listserv, go to: >>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >>> ***** >>> >>> 30x30x1 nm, 7 colors ... see figure 1 for colors - the 7 'colors' can >>> probably be extended to more for a bit more money. >>> http://www.nature.com/nature/journal/v481/n7382/extref/nature10734-s1.pdf >>> >>> The same lab has the adjacent article - >>> http://www.nature.com/nature/journal/v481/n7382/extref/nature10745-s1.pdf >>> >>> >>> 30x30x1 nm (yes, nanometer) is very nice - perhaps the fluorescence >>> nanoscope developers should spend a bit less time on pushing from the >>> current ~20x20x50 nm (which should get to 10x10x25 nm by a certain >>> improvement in light output while on) and spend a bit more time on >>> producing making new biomedical discoveries. >>> >>> I may say a few words about this approach during my short talk at the >>> ABRF nanoscopy session in Orlando in late March. >>> I might start the talk with the nice "SnapShot; light Microscopy" in >>> Cell (Nov 23, 2011 issue - I was able to download from home) >>> >>> http://download.cell.com/pdf/PIIS0092867411013535.pdf?intermediate=true >>> >> >> > > -- ------------------------------------------------------------ Steffen Dietzel, PD Dr. rer. nat Ludwig-Maximilians-Universität München Walter-Brendel-Zentrum für experimentelle Medizin (WBex) Head of light microscopy Mail room: Marchioninistr. 15, D-81377 München Building location: Marchioninistr. 27, München-Großhadern |
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