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Cell Biology JoVE (Open Access) Citable · DOI

Multilayer Mounting for Long-term Light Sheet Microscopy of Zebrafish

DOI: 10.3791/51119-v
What you'll learn
  • ✓ Prepare fluorinated ethylene propylene tubes for embedding
  • ✓ Mount zebrafish embryos in low-concentration agarose
  • ✓ Layer samples for long-term light sheet imaging
  • ✓ Acquire multi-day images of developing zebrafish
Protocol

Biopharma Insights The development of zebrafish can be followed over days with light sheet microscopy when embryos are embedded in optically clear polymer tubes with low-concentration agarose.

Difficulty
intermediate
Total time
~1-2 hours per sample (excluding imaging duration)
Model organism
Zebrafish (Danio rerio)
Biosafety
BSL-1

Steps

1
Prepare fluorinated ethylene propylene tubes

Cut and prepare FEP tubes that will hold the agarose-embedded zebrafish for imaging.

▶ 01:55
2
Mount zebrafish in multilayer agarose

Embed zebrafish embryos in low-concentration agarose within the prepared tubes using a multilayer mounting approach.

▶ 03:02
3
Acquire multi-day zebrafish images

Perform light sheet microscopy to capture images of the developing zebrafish over multiple days.

▶ 06:26
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