X-Message-Number: 29396 Date: Fri, 06 Apr 2007 18:19:42 -0400 From: Keith Henson <> Subject: How to make a brain transparent Forwarded from an EP group. Keith How to make a brain transparent 16:26 02 April 2007 NewScientist.com news service Roxanne Khamsi The new ultramicroscopy technique images a whole mouse brain in 3D (Image: Hans-Ulrich Dodt et al./Nature Methods) The entire neural network of a mouse's brain has been seen in 3D for the first time, using a new technique that renders tissues transparent. The method - dubbed "ultramicroscopy" - has also enabled researchers to visualise the detailed anatomy of a mouse embryo in 3D. It will provide new insight into how organs such as the brain develop, the researchers say. Until now, it has been impossible to visualise entire neuronal networks in an intact brain - techniques such as computer tomography (CT scans) or magnetic resonance imaging (MRI) do not have the resolution to reveal detail at the cellular level. Slicing the brain for microscopic imaging is possible, but creating a 3D image from many slices is laborious and prone to distortion problems. Hans-Ulrich Dodt, now at Vienna University of Technology in Austria, and colleagues, have combined two old techniques to make a new tool that allows researchers to look at an entire brain on a microscopic level. Take a look at a selection of images and video clips of the process here. The new technique can also be used to image whole mouse embryos (Image: Hans-Ulrich Dodt et al./Nature Methods) Refractive index Using rodents genetically engineered to produce florescent molecules in their nerve cells, the team extracted whole mouse brains and submerged them in alcohol to flush the water out of the tissues. The dehydrated brains were then placed into an oil mixture containing the solvents benzyl-benzoate and benzyl-alcohol. Importantly, this medium has exactly the same light refractive index as protein - meaning that any light passing through the medium would continue to pass through the brain tissue at the same angle. Usually, when light enters a body tissue, it is scattered by the different refractive index of the tissue, in the same way that light is bent as it passes through water, making submerged items appear distorted. The medium effectively made the organ transparent, much like a drop of oil on a piece of paper can make light pass more easily through the page, Dodt explains. A whole mouse brain showing individual neurons fluorescing (Image: Hans Ulrich Dodt) Nerve connection The next step involved viewing cross-sections of the brain by shining a thin sheet of light through the organ. As this sliver of light about six micrometres thick passed through the brain, it caused all of the neurons in its path to fluoresce. A computer then integrated the images obtained from scanning the thin sheet of light across the brain to give a 3D picture of how the nerves connect (see the results pictured, lower right) Dodt claims the technique is a significant advance over previous methods to image the brain. Typically, approaches have involved physically cutting the brain into thin slices, and then staining the neurons in each slice. But the act of physically slicing the organ can distort the position of the nerves, he explains. By comparing the scans of mouse embryos with those of adult mice they hope to get a better view on how mammalian brain networks change during development. This could give new insight into how mammalian brains change over time and what happens to information networks as a result of disease. Journal reference: Nature Methods (DOI: 10.1038/nmeth1036) Source: NewScientist http://www.newscientisttech.com/article/dn11518?DCMP=NLC-nletter&nsref=dn11518 Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=29396