Fluorescent-protein stabilization and high-resolution imaging of cleared, intact mouse brains.

Fluorescent-protein stabilization and high-resolution imaging of cleared, intact mouse brains.

Blog Article

In order to observe and quantify long-range neuronal connections in intact mouse brain by light microscopy, it is first necessary to clear the brain, thus suppressing refractive-index variations.Here we describe a method that clears the brain and preserves the signal from proteinaceous fluorophores using a pH-adjusted non-aqueous index-matching medium.Successful clearing is enabled through the use of either 1-propanol or tert-butanol during dehydration whilst maintaining a basic pH.

We show that high-resolution fluorescence imaging of pure energy jeans entire, structurally intact juvenile and adult mouse brains is possible at subcellular resolution, even following many months in clearing solution.We also show that axonal long-range projections that are EGFP-labelled by modified Rabies virus can be imaged throughout the brain using a purpose-built light-sheet fluorescence microscope.To demonstrate the viability of the technique, we determined a detailed map of the monosynaptic projections onto a target cell population in the lateral entorhinal cortex.

This example demonstrates that our skull bride and groom method permits the quantification of whole-brain connectivity patterns at the subcellular level in the uncut brain.