3D Movies of Living Cells

This is amazing stuff.  We now have a method to map cells in threedimensions.  Add different wavelengthsand the field of such imaging is entering a complete new era of development andgeneral importance.

There are bunch of images herewell worth looking at.  Go for it.

This methodology will provide usa major refinement on classical results and provide images that confirm wellaccepted knowledge often not well pictured. I suspect that a lot of old time frustration just got fixed.

New microscope captures 3D movies of living cells

By Paul Ridden

04:14 March 15, 2011

A new kind of microscope has been invented that is able tonon-invasively take a three dimensional look inside living cells with stunningresults (Images courtesy of Betzig, Planchon, and Gao)

http://www.gizmag.com/3d-microscope-movies-living-cells/18138/

In some cases, looking at a living cell under a microscope can cause itdamage or worse, can kill it. Now, a new kind of microscope has been inventedby researchers from the Howard Hughes Medical Institute that is able tonon-invasively take a three dimensional look inside living cells with stunningresults. The device uses a thin sheet of light like that used to scansupermarket bar codes and could help biologists to achieve their goal of understandingthe rules that govern molecular processes within a cell.

Veteran microscope innovator Eric Betzig says that the field ofmicroscopy has been hindered by the fact that many techniques require cells tobe killed and fixed before being viewed. Light produced by microscopes used forlive-cell techniques can, in some cases, actually cause damage to the cells.The light also floods the whole area being examined, not just the small portionthat's in focus – producing blur from the out-of-focus regions.

Two years after arriving at HHMI's Janelia Farm Research Campus, Betzig started workingways to overcome these problems.

"The question was, is there a way of minimizing the amount ofdamage you're doing so that you can then study cells in a physiological mannerwhile also studying them at high spatial and temporal resolution for a longtime?" said Betzig.

First developed around a 100 years ago, plane illuminationmicroscopy involves shining light through the side of a sample rather than fromthe top. While offering some promise, Betzig's group found that thetechnique still exposed too much of the sample. A much thinner sheet of lightwas produced using by sweeping a Bessel beam – a kind of non-diffracting lightbeam – across the sample but the light produced by this form of planeillumination microscopy proved to be somewhat weak, making the pattern of illuminationlook somewhat like a bullseye.

Working with postdoctoral researchers Thomas Planchon and Liang Gao,Betzig has spent the last couple of years refining the process to try andovercome the problem. First, instead of sweeping the Bessel beam across thesample, the group rapidly switched it off and on – a method known asstructured illumination. Then by concentrating the light to a narrow centralpart of the Bessel beam using something called two-photon microscopy, theywere able to build 3D stacks of the sample at nearly 200 images per second togenerate movies of processes like cell division in stunning detail.

Betzig says that Bessel beam plane illumination microscopy will prove apowerful tool for cell biologists, since it non-invasively images the rapidlyevolving three-dimensional complexity of cells.

The research is described in detail in a paper entitled Rapid three-dimensional isotropic imaging of living cells usingBessel beam plane illumination, which was recently published in thejournal Nature Methods.