Home Cell Biology Imaging Cleared Intact Biological Systems at a Cellular Level by 3DISCO
Cell Biology JoVE (Open Access) Citable · DOI

Imaging Cleared Intact Biological Systems at a Cellular Level by 3DISCO

DOI: 10.3791/51382-v
What you'll learn
  • Apply 3DISCO tissue clearing protocol using sequential organic solvents
  • Image cleared intact organs at cellular resolution without sectioning
  • Interpret representative fluorescence images from transparent biological specimens
Protocol

To obtain high-resolution images of fluorescently labeled cells within large tissues, ideally, the biological samples should be imaged without sectioning. 3DISCO is a straightforward tissue clearing procedure based on sequential incubation with organic solvents. Upon clearing, the organs become transparent allowing an end-to-end laser scan of the specimen.

Difficulty
intermediate
Total time
~2–3 days per organ (including solvent incubation steps)

Steps

1
Image cleared organs using laser scanning

Acquire high-resolution fluorescence images of transparentized tissues via end-to-end laser scanning to visualize labeled cells throughout intact organs without sectioning.

▶ 01:32
2
Examine representative 3DISCO-cleared tissue images

Review sample fluorescence micrographs of cleared tissues demonstrating cellular-level visualization achieved by the 3DISCO transparency protocol.

▶ 04:14
3
Summarize tissue clearing and imaging workflow

Consolidate key principles: sequential organic solvent incubation renders organs transparent, enabling label-free whole-organ imaging at cellular resolution.

▶ 06:54
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