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Microscopy & Imaging JoVE (Open Access) Citable · DOI

Analysis of Zebrafish Kidney Development with Time-lapse Imaging Using a Dissecting Microscope Equipped for Optical Sectioning

DOI: 10.3791/53921-v
What you'll learn
  • ✓ Set up time-lapse imaging of zebrafish kidney development using fluorescence dissecting microscopy
  • ✓ Perform microinjection and mounting procedures for live embryo imaging
  • ✓ Correct focal and planar drift during long-term optical sectioning experiments
  • ✓ Distinguish normal from disturbed kidney development patterns in zebrafish
Protocol

The method described here allows time-lapse analysis of organ development in zebrafish embryos by using a fluorescence dissecting microscope capable of performing optical sectioning and simple strategies of readjustment to correct focal and planar drift.

Difficulty
advanced
Total time
~4–6 hours (embryo collection through imaging session)
Model organism
Zebrafish (Danio rerio) embryos
Biosafety
BSL-1

Steps

1
Collect and prepare fertilized zebrafish eggs

Obtain fertilized eggs from breeding tanks and prepare them for subsequent microinjection and imaging. Ensure proper staging and health of embryos before proceeding.

▶ 01:17
2
Microinject developing zebrafish embryos

Perform microinjection into early-stage embryos to introduce fluorescent markers or molecular probes for visualization of kidney development.

▶ 02:27
3
Mount embryos for live fluorescence imaging

Prepare and secure injected embryos in an imaging chamber with appropriate mounting medium to enable sustained time-lapse microscopy.

▶ 04:21
4
Compensate for focal and planar drift

Apply optical sectioning and readjustment strategies to maintain image quality and correct for specimen drift during extended time-lapse acquisition.

▶ 06:59
5
Analyze normal and disturbed kidney development

Review and interpret time-lapse image sequences to identify developmental landmarks and distinguish morphological changes in wild-type versus experimentally perturbed embryos.

▶ 08:04
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