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PLoS One. 2017 Jul 7;12(7):e0180486. doi: 10.1371/journal.pone.0180486. eCollection 2017.

Ribbon scanning confocal for high-speed high-resolution volume imaging of brain.

Author information

1
Center for Vaccine Research, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
2
Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
3
Department of Neurobiology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
4
Center for Biologic Imaging and the Department of Cellular Biology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
5
Department of Immunology, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.

Abstract

Whole-brain imaging is becoming a fundamental means of experimental insight; however, achieving subcellular resolution imagery in a reasonable time window has not been possible. We describe the first application of multicolor ribbon scanning confocal methods to collect high-resolution volume images of chemically cleared brains. We demonstrate that ribbon scanning collects images over ten times faster than conventional high speed confocal systems but with equivalent spectral and spatial resolution. Further, using this technology, we reconstruct large volumes of mouse brain infected with encephalitic alphaviruses and demonstrate that regions of the brain with abundant viral replication were inaccessible to vascular perfusion. This reveals that the destruction or collapse of large regions of brain micro vasculature may contribute to the severe disease caused by Venezuelan equine encephalitis virus. Visualization of this fundamental impact of infection would not be possible without sampling at subcellular resolution within large brain volumes.

PMID:
28686653
PMCID:
PMC5501561
DOI:
10.1371/journal.pone.0180486
[Indexed for MEDLINE]
Free PMC Article

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