Probing:
The non-invasive probing of zebrafish embryos can be conducted at various stages of development.
For early stages dechorionation of the embryos in embryo medium (zebrafish book) and placing the embryos on a
small dish is sufficient. In the later stages where periodic tail movement has begun, dechorionation followed
by 1-2 drops of tricaine solution will stabilize the embryo.
A probe reading from >100 micrometers from the embryo is essential for proper calibration. The probe can then be
placed near the edge of the embryo where grid scanning can be conducted. The data from the scans can be exported to text
and analyzed with kinemage and surface maps.
Procedure for using Janus Green B to find active cells:
Prepare Janus Green B solution by adding 3mg of Janus Green B to 30ml
of isopropanol. Place 1-2 drops on a slide or dish and let the alcohol
evaporate. Place embryon in medium (pond water) onto the dish. Let sit
at room temperature and observe under microscope.
Tricaine Anesthetic:
Prepare tricaine stock solution by adding 400mg of tricaine powder to 97.9ml
of distilled water. Add ~2.1ml of Tris (ph9) to adjust the ph to 7. Store in the freezer.
To prepare the tricaine solution, add 4.2ml of tricaine stock to 100ml of tank water.
To use, drop fish into beaker of tricaine solution and remove when gill movement
has slowed to one breath per second.