Eye Lens Dissection: A Technique to Observe Zebrafish Lens Morphology

Abstract

Source: Vorontsova I. et al, Assessment of Zebrafish Lens Nucleus Localization and Sutural Integrity. J. Vis. Exp. (2019).

This video describes the protocol which is developed to dissect adult and larval zebrafish lenses to analyze cortical lens morphology and structural integrity of the lens sutures.

Protocol

1. Dissection of Larval and Adult Zebrafish Lenses

1. Anesthetize fish from 6 dpf larvae to adulthood with tricaine until non-responsive to touch but still showing a strong heartbeat. Measure fish standard length as per Schilling (2002) for staging.
2. Immediately excise eyes using micro-dissection scissors and place into a dissection dish in PBS (Figure 1 shown for adult eyes).
   1. Make a custom 35 mm dish filled with silicone for lens dissections. Once silicone has set, excise a divot around 2-3 mm in diameter and 0.5 mm deep for immobilizing adult eyes/lenses during dissection.
   ....

Results

Figure 1: Zebrafish lens dissection. (A) Eyes are dissected from anesthetized fish and placed posterior side up (B). (C) Eyes are immobilized by forceps via the optic disc (od) and two to.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Phosphate buffered saline (PBS)	Fisher Scientific	BP399
Glass bottom microwell dish (35mm Petri dish, 14 mm microwell, #1.5 coverglass)	MatTek Corporation	P35G-1.5-14-C
Dumont #5 forceps	Dumont & Fils		Keep forceps sharpened
Vannas micro-dissection scissors	Ted Pella Inc	1346	Sharp/sharp straight tips
Confocal microscope	Nikon		Eclipse Ti-E
Sodium Hydroxide beads	Fisher Scientific	S612-3	CAUTION - corrosive/irritating to eyes and skin, target organ - Journal of Visualized Experiments www.jove.com Copyright © 2019 Journal of Visualized Experiments Page 2 of 2 respiratory system, corrosive to metals
Disposable Pasteur glass pipets	Fisherbrand	13-678-20A