This video demonstrates the dissection and preparation of a flat mount of the adult zebrafish kidney. The prepared flat mount can be visualized under the microscope for specimen analysis.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Dissection and Flat Mount Preparation of the Adult Zebrafish Kidney from an Unfixed Animal Sample to Visualize Fluorescent Dextran Labeling of the Proximal Tubule Euthanize the dextran-injected adult zebrafish by placing it into a dish with 0.2% Tricaine pH 7.0 for 4–5 min. NOTE: Ensure tha…
Representative Results
Figure 1. Adult zebrafish kidney flat mount preparation and application to visualize conjugated dextran uptake in the PCT segment of kidney nephrons. (A) Brightfield image of a kidney specimen flat mount preparation, in which the organ has been positioned flat on a glass slide, with a coverslip placed on top of the tissue that is resting on four divots of modeling clay (hot pink color)….
Divulgaciones
The authors have nothing to disclose.
Materials
1X PBS
Made by diluting 10 X PBS in distilled water.
Fine Forceps
Roboz
RS-1050
Dumont Tweezers Pattern #55
Glass slide
Thermo-Fisher
4445
White Frost
Glass coverslip
Thermo-Fisher
12-540A
18 x 18 mm
Modeling clay
Hasbro
Playdoh
Other modeling clays can be substituted and work similarly.
Filter set used was as follows: Hoechst/DAPI filter was used for DAPI, propidium iodide, dextran-cascade blue and alkaline phosphatase detection. GFP/FITC filter was used for dextran-FITC, dextran lucifer yellow, and antiGFP detection. The TRITC filter was used for dextran-fluoro-ruby, PI, and DBA detection.
Filter set used was as follows: Hoechst/DAPI filter was used for DAPI, propidium iodide, dextran-cascade blue and alkaline phosphatase detection. GFP/FITC filter was used for dextran-FITC, dextran lucifer yellow, and antiGFP detection. The Texas Red filter was used for dextran-fluoro-ruby, PI, and DBA detection.
Kidney Flat Mount Preparation: A Procedure to Dissect and Prepare a Flat Mount for Microscopic Analysis of Adult Zebrafish Kidney. J. Vis. Exp. (Pending Publication), e20573, doi: (2023).