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

Area of Science:

• Developmental Biology
• Microscopy Techniques
• Zebrafish Models

Background:

• Understanding organogenesis requires detailed recording of spatial and temporal tissue development.
• Zebrafish embryos are a valuable model for studying vertebrate development due to their optical transparency and rapid external development.

Purpose of the Study:

• To describe a novel, accessible time-lapse microscopy method for analyzing kidney organogenesis in zebrafish embryos.
• To demonstrate the method's utility in observing both normal and impaired kidney development.

Main Methods:

• Utilized transgenic zebrafish (Tg(wt1b:GFP)) with fluorescently labeled kidney structures.
• Employed a fluorescence dissecting microscope with structured illumination and z-stack acquisition capabilities.
• Induced renal defects by injecting antisense morpholino oligonucleotides against the Wilms tumor gene wt1a.

Main Results:

• Successfully performed time-lapse analysis of normal and genetically impaired zebrafish kidney development.
• The method incorporated an autofocus strategy to counteract focal drift and relocation grids to correct xy-drift.
• Demonstrated the advantages of a zoom microscope, including a high depth of field and extended working distance, for organogenesis studies.

Conclusions:

• The described method offers a practical and cost-effective alternative to complex microscopy setups for time-lapse recording of developing tissues.
• This technique facilitates the study of organogenesis and organ dynamics in zebrafish, applicable even with fluorescence stereo microscopes lacking optical sectioning.
• The approach is valuable for researchers studying kidney development and other organogenesis processes.