A Scalable Staining Strategy for Whole-Brain Connectomics.
Xiaotang LuYuelong WuRichard L SchalekYaron MeirovitchDaniel Raimund BergerJeff William LichtmanPublished in: bioRxiv : the preprint server for biology (2023)
Mapping the complete synaptic connectivity of a mammalian brain would be transformative, revealing the pathways underlying perception, behavior, and memory. Serial section electron microscopy, via membrane staining using osmium tetroxide, is ideal for visualizing cells and synaptic connections but, in whole brain samples, faces significant challenges related to chemical treatment and volume changes. These issues can adversely affect both the ultrastructural quality and macroscopic tissue integrity. By leveraging time-lapse X-ray imaging and brain proxies, we have developed a 12-step protocol, ODeCO, that effectively infiltrates osmium throughout an entire mouse brain while preserving ultrastructure without any cracks or fragmentation, a necessary prerequisite for constructing the first comprehensive mouse brain connectome.
Keyphrases
- resting state
- electron microscopy
- functional connectivity
- white matter
- high resolution
- cerebral ischemia
- randomized controlled trial
- induced apoptosis
- magnetic resonance imaging
- multiple sclerosis
- prefrontal cortex
- blood brain barrier
- computed tomography
- magnetic resonance
- cell proliferation
- mass spectrometry
- brain injury
- cell death
- living cells
- flow cytometry
- combination therapy
- pi k akt