Cleared brain

 Axonal projections from the thalamus to layer IV in the mouse barrel cortex.

Scale and CLARITY methods render biological samples optically transparent but completely preserves fluorescent signals in the structures. In the treated mouse brain, neurons labeled with genetically encoded fluorescent proteins, can be visualized at an unprecedented depth in millimeter-scale networks and at subcellular resolution. The improved depth and scale of imaging permits comprehensive three-dimensional reconstructions of brain sections such as cortex, corpus callosum or hippocampus. The extent was limited only by the working distance of the objective lenses. These methods are useful for light microscopy-based connectomics of cellular networks in brain and other tissues

Using the appropriate process, transparent brain tissue is suitable for deep imaging with the Olympus Scaleview objective family to up to a depth of more than 4 mm (illuminated by a two-photon laser and using GaAsP detectors).

XZ projection of an image stack of cells and axonas in the mouse barrel cortex.

Images courtesy of:

Gabriel Davis Jones, Dr. Kay Richards & Dr. Steven Petrou
Florey Institute of Neuroscience and Mental Health, Melbourne, Australia

Using the Scale imaging technique with the Olympus Scaleview 25x/1.05 objective at 1040nm using Femto 3D - RC microscope.
(used Huygens software to deconvolve and Amira to stitch and project.)