Optimized methods to image hepatic lipid droplets in zebrafish larvae.

Area of Science:

• Comparative biology
• Cell biology
• Zebrafish models

Background:

• Zebrafish larvae's optical transparency facilitates in vivo visualization of subcellular structures.
• Lipid droplets (LDs) are crucial cellular components, and their accumulation in hepatocytes signifies liver disease.
• Zebrafish are extensively utilized in research on lipid biology and liver pathologies.

Purpose of the Study:

• To comprehensively analyze methods for investigating LD distribution and dynamics in zebrafish larval livers.
• To compare the efficacy of various lipid dyes and transgenic reporters for LD visualization.
• To provide a resource for researchers studying LD biology in zebrafish disease models.

Main Methods:

• Comparative analysis of lipid dyes: Oil Red O, Nile Red, LipidTox, and LipidSpot.
• Utilization of transgenic reporter lines expressing EGFP-fused perilipin 2 (PLIN2) on LDs.
• Application of microscopy techniques from stereomicroscopy to confocal imaging for LD analysis.

Main Results:

• Demonstration of the strengths and limitations of each lipid visualization method.
• Establishment of protocols for quantifying hepatic LD size, number, and dynamics.
• Adaptability of methods for both live and fixed zebrafish larvae, across various imaging resolutions.

Conclusions:

• Multiple validated methods exist for assessing hepatic LDs in zebrafish.
• These protocols enable detailed cellular-level analysis of LDs in physiological and pathological states.
• This comparative study serves as a valuable guide for researchers utilizing zebrafish models for liver disease investigation.