This material is from the 4th edition of The Zebrafish Book. The 5th edition is available in print and within the ZFIN Protocol Wiki.


Gelatin Embedding for Vibratome Sectioning of Embryos or Larvae

(Source: R. BreMiller)

1. Fix fish as desired.

2. Wash in fix buffer, 3 times for 5 min each wash.

3. Soak fish in 0.3 M sucrose for at least 30 min. Remove yolk sac from fish with forceps.

4. Transfer fish with as little sucrose solution as possible into 17% gelatin in 10% Hank's saline at 37C.

5. Swirl the fish to coat it well with gelatin and transfer it with a drop of gelatin onto a 5 mm square piece of glass made by scoring and breaking a clean microscope slide.

6. Orient the animal under a dissecting microscope with fine wire or forceps. The process may be facilitated by warming or cooling the glass plate.

7. Fasten the plate to a vibratome chuck with cyanoacrylic glue and clamp the chuck in the vibratome.

8. Surround the tissue with cold 0.3 M sucrose. Use frozen cubes of sucrose to maintain the temperature.

9. Sections can be cut 25-40 um thick with this technique. Pick them up with brush and transfer to cold, subbed slides.

10. Remove excess sucrose with a Kimwipe and allow the slides to dry. They should be processed within a few hours.

Gelatin embedding

The Zebrafish Book