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Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain
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Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain
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Journal Article
Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain
2011
Overview
The authors describe a chemical approach for imaging deep into fixed brain tissue using Sca
l
e, a solution that renders biological samples transparent, but preserves fluorescent signals. This technique allows for imaging at unprecedented depth and at subcellular resolution, and makes three-dimensional reconstruction of neural networks possible without serial sectioning.
Optical methods for viewing neuronal populations and projections in the intact mammalian brain are needed, but light scattering prevents imaging deep into brain structures. We imaged fixed brain tissue using Sca
l
e, an aqueous reagent that renders biological samples optically transparent but completely preserves fluorescent signals in the clarified structures. In Sca
l
e-treated mouse brain, neurons labeled with genetically encoded fluorescent proteins were visualized at an unprecedented depth in millimeter-scale networks and at subcellular resolution. The improved depth and scale of imaging permitted comprehensive three-dimensional reconstructions of cortical, callosal and hippocampal projections whose extent was limited only by the working distance of the objective lenses. In the intact neurogenic niche of the dentate gyrus, Sca
l
e allowed the quantitation of distances of neural stem cells to blood vessels. Our findings suggest that the Sca
l
e method will be useful for light microscopy–based connectomics of cellular networks in brain and other tissues.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ Animal Genetics and Genomics
/ Animals
/ Biomedical and Life Sciences
/ Brain - growth & development
/ Imaging, Three-Dimensional - methods
/ Intermediate Filament Proteins - metabolism
/ Luminescent Proteins - genetics
/ Luminescent Proteins - metabolism
/ Mice
/ Microscopy, Fluorescence - methods
/ Nerve Tissue Proteins - metabolism
/ Nestin
/ Neural Cell Adhesion Molecule L1 - metabolism
/ Neural Stem Cells - physiology
/ Proteins
/ Reagents
/ Receptors, AMPA - metabolism
/ Sucrose
