Kubke Lab:Research/CND/Records2010-2011Summer/RC005

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Cranial Nerve Development Experiment

Embryo details


Species: Gallus gallus domesticus
Embryo Name: RC005
Embryo stage: ST19 Fabiana staged this embryo on the 25/1 and confirmed it was a ST19. --Malisha Hettiarachchi 15:22, 27 January 2011 (EST)


Staging description: Originally staged by Reuben as a ST20, but Fabiana restaged on 25/1as a ST19 and Fabiana according to the Hamburger and Hamilton (1951) staging system. The description below corresponds to Reuben's staging.

  • Wing and leg buds barely formed.
  • The leg is slightly longer than the wing.
  • Allantois has vesicularized and is roughly the same size as the midbrain (indicative of ST20).
(Note: what do you mean by vesicularised? --MF Kubke 00:39, 27 January 2011 (EST))
(Note: This is how H&H worded it. I thought they were describing the spherical/vesicle-like form that it takes on at this stage and since the embryo's allantois looked the same as their images I chose to use the same term.)

(see Notebook entry 10th January 2011)
Fixation:
Cryoprotection: Was stored in PFA, staged in .9% saline and then replenished with PFA.
Material label and storage:Stored in vial filled with PFA labeled RC005 and placed in a container labeled 'Reuben's staged embryos' next to the dissection microscope.

Experiment details


Objective:To use as a practice embryo for staging.
Procedure: Staged the embryo by observing the embryo under a dissection microscope and comparing its morphological properties with those described in the Hamburger and Hamilton (1951) paper whilst looking at our own compilation of images organized by stage.
Comments:


Cryostat Sectioning Cryostat settings (for a more detailed protocol visit Kubke_Lab:Cryomicrotomy)

Cryostat Histology Lab
Knife
Day Cut 27/11
Knife Angle 0
Chamber Temp -19C
Object Temp -19c
Glass Slides Gelatin Subbed
Plane of section Saggital
Number of slides 9
Observations The sections "jumped" onto the slide and the whole embryo was

(Include in your observations, eg ,were the sections serial, was any section lost, was quality assessed, etc)



Results

Embryo is approximately ST20.

Images

Summary

This embryo may be used in further experiments as approximately a ST20 however confirmation is needed.


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