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3D Structured Illumination Microscopy

JJackson

In Memoriam - Senior Moderator
I found this on the Science.com site and I think it is an interesting idea. Different parts of the electro-magnetic spectrum are useful for imaging objects on different scales. The maximum resolution is a function of the wavelenght so for very small objects to resove detail you need short wavelengths but short wavelength means hi frequency and more energetic radiation. So anything that allows for improved resoultion of delicate organelles without blasting them with ever more distructive radiation is a good thing.

Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy
Lothar Schermelleh,1* Peter M. Carlton,2* Sebastian Haase,2,4 Lin Shao,2 Lukman Winoto,2 Peter Kner,2 Brian Burke,3 M. Cristina Cardoso,4 David A. Agard,2 Mats G. L. Gustafsson,5 Heinrich Leonhardt,1*
dagger.gif
John W. Sedat2*
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Fluorescence light microscopy allows multicolor visualization of cellular components with high specificity, but its utility has until recently been constrained by the intrinsic limit of spatial resolution. We applied three-dimensional structured illumination microscopy (3D-SIM) to circumvent this limit and to study the mammalian nucleus. By simultaneously imaging chromatin, nuclear lamina, and the nuclear pore complex (NPC), we observed several features that escape detection by conventional microscopy. We could resolve single NPCs that colocalized with channels in the lamin network and peripheral heterochromatin. We could differentially localize distinct NPC components and detect double-layered invaginations of the nuclear envelope in prophase as previously seen only by electron microscopy. Multicolor 3D-SIM opens new and facile possibilities to analyze subcellular structures beyond the diffraction limit of the emitted light.
[SIZE=-1]1 Center for Integrated Protein Science, Department of Biology, Ludwig Maximilians University Munich, 82152 Planegg-Martinsried, Germany.
2 Department of Biochemistry and Biophysics, University of California, San Francisco, CA 94143, USA.
3 Department of Anatomy and Cell Biology, University of Florida, Gainesville, FL 32610, USA.
4 Max Delbr?ck Center for Molecular Medicine, 13125 Berlin, Germany.
5 Department of Physiology and Program in Bioengineering, University of California, San Francisco, CA 94143, USA. [/SIZE]
* These authors contributed equally to this work.
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To whom correspondence should be addressed. E-mail: h.leonhardt@lmu.de (H.L.); sedat@msg.ucsf.edu (J.W.S.)









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Re: 3D Structured Illumination Microscopy

reminds me to another article in German which I read recently.
I couldn't find any STED-pictures :-(


Max Planck Institute for biophysical chemistry
New microscope makes sharp 3-D-view possible into the cell
Goettingen. DPA/baz. Researchers of the Max Planck Institute for biophysical
chemistry in Goettingen, according to own reports, reached a "Milestone in the Nanoskopie".
With far improved optical microscope are for the first time three-dimensional views into living
cells with a dissolution of under 45 nanometers (millionth millimeters) possible,
communicated Institut on Thursday.
The so-called STED microscope developed by award winner Stefan Hell had made
a such dissolution possible so far only in two directions in space. With the new
microscope one now can examine deeply in the cell inside hidden proteins in
three dimensions. Recently the researchers had reported of a resolving power
of approximately 60 nanometers. How the university communicated Goettingen
at the beginning of May, the researchers want to examine the chemical procedures
of the nerve cells around Silvio Rizzoli with the microscope, in order to find
causes for neurological and psychological disturbances.


http://www.nature.com/nature/journal/v439/n7075/full/nature04378.html

http://www.virologyj.com/content/4/1/49

http://www.jcb.org/cgi/reprint/91/3/601.pdf
 
Re: 3D Structured Illumination Microscopy

while searching for more pictures, I found this:

http://www.denniskunkel.com/index.php?cPath=13&sort=1a&page=5

I knew, they are polymorphic, can be spherical or filamentous,
but these irregular-looking flu-viruses are unexpected and hard
to understand for me.



lots of pictures with google - images search (influenza microscope)
I liked these:

http://www.greaterimmunity.com/Images/Before and after Influenza A.jpg
http://www.medscape.com/content/2000/00/40/84/408402/art-mns1228.bark.fig1.gif
http://images.google.com/imgres?img...pe&start=60&gbv=2&ndsp=20&hl=en&sa=N&ie=UTF-8
http://images.google.com/imgres?img...mages?q=schermelleh&gbv=2&hl=en&sa=G&ie=UTF-8

http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=2112819&blobtype=pdf
 
Re: 3D Structured Illumination Microscopy

WAOUH ! thank you Jackson.. I ve heard about it and I was searching.
 
Re: 3D Structured Illumination Microscopy

The fetal looking DKNIH1 is a close up DKNIH2 (central viron) and is the only one of that shape on the plate but yes they do seem to come in a range of shapes.
Wikipedia had something on STED and says it is an implementation of RESOLFT the equations for which were developed at the Max Plank Institute.
 
Re: 3D Structured Illumination Microscopy

Now if you like microscopes check this out.

A lecture on electron holography by Akira Tonomura recorded at the Royal Society in London 1994.

I kid you not. They have a movie of them herding self annihilating microscopic magnetic vortices on the surface of a super-conducting magnet. (If you have seen footage of the magnetic vortices on the Sun's surface cancelling each other out it is a bit like that)
You may want to skip the first half unless you want a lecture on harmonics and interference patterns. The interesting stuff is at the end.

gsgs I believe you are a mathematician? If so do you know of Paul Erdos his biographer Paul Hoffman gave a lecture which can also be found on this site. It is more on the eccentricities of the man than on his work but is definitely worth watching. The site also host a copy of Feynman's Auckland QED lecture and he is always worth watching.
 
Re: 3D Structured Illumination Microscopy

someone knows how to copy the describing text into the .gif - images ?
Unfortunately this is rarely found, so when you save the picture you lose
the description. We need it in text and included in the picture, there should
be a conversion utility.
There should also always be a scale-mark on these microscope pictures,
even if it is unsure, possibly inexact.
There should also be a google-image-search which searches for non-copyright
images only.

Thanks for the link, I tried it, but couldn't easily find the interesting things,
maybe I try again later.
I don't like long videos, they eat computer memory, are also often difficult to
understand for me. I do like short summaries, which can be extended
to longer summaries if wanted. At this point I don't want to understand,explore
microscope technics so much but rather look at the results, the (virus-) pictures ;-)

Yes I heard some stories about Erdos in the math-forums, they say he just
often worked for food and lodging. His most famous quote :
> a mathematician is a device which turns coffee into theorems.
mathematics is so beautiful but often not so useful.
Feynman,Erdos are interesting for historians for scientists their results
are interesting only.
 
Re: 3D Structured Illumination Microscopy

"someone knows how to copy the describing text into the .gif - images ?"

I have an old utility called CorelCapture - which came as part of the CorelDraw suite. While I have little use for the main program this utility is great. I have it set up so if I press the [F7] key it changes my cursor and I can drag a rectangle over the object of interest. This image is then saved to clipboard and/or to a file called capture.jpg which is easy to then upload into posts. It means not having to mess around with cropping images and some journals make their data available using .pdf s of scans of the printed page which can not therefore be copied and pasted.
 
Re: 3D Structured Illumination Microscopy

Thanks Safran
The optical/SIM comparison stills show how that little bit of extra resolution helps.
 
Re: 3D Structured Illumination Microscopy

can we expect such rotating influenza-virus videos
which show the internal structures (RNPs) ?
 
50 nm resolution with a white-light microsope

50 nm resolution with a white-light microsope

Here is another workaround for by passing the wavelength limitations I referred to in the threads opening post.<o:p></o:p>
<o:p> </o:p>
<o:p> </o:p>
Optical virtual imaging at 50 nm lateral resolution with a white-light nanoscope.
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Abstract
The imaging resolution of a conventional optical microscope is limited by diffraction to ~200 nm in the visible spectrum. Efforts to overcome such limits have stimulated the development of optical nanoscopes using metamaterial superlenses, nanoscale solid immersion lenses and molecular fluorescence microscopy. These techniques either require an illuminating laser beam to resolve to 70 nm in the visible spectrum or have limited imaging resolution above 100 nm for a white-light source. Here we report a new 50-nm-resolution nanoscope that uses optically transparent microspheres (for example, SiO2, with 2 μm<diameter><9 μm) as far-field superlenses (FSL) to overcome the white-light diffraction limit. The microsphere nanoscope operates in both transmission and reflection modes, and generates magnified virtual images with a magnification up to ×8. It may provide new opportunities to image viruses and biomolecules in real time.

Edit:
The BBC website also have an article on this.
http://www.bbc.co.uk/news/science-environment-12612209

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Re: 3D Structured Illumination Microscopy

This is even better checkout Live 3D STORM this is unbelievably clever.
I am afraid it is 50mins long and only the first of four parts.
If you are not interested in the physics of the system you could just skip to near the end and watch the protein molecules moving around the cell but it really is worth the 50mins.
I was never expecting to be able to see a clathrin pit - and its contents - moving around a live cell using light microscopy at a resolution of 25nm in the xy plane and 50nm along the z axis, individual molecules positions are resolved to 2nm.
 
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