Kubke Lab:Research/CND/Records2010-2011Summer/MH006
Cranial Nerve Development | Experiment |
Embryo details
Species:Gallus gallus domesticus
Embryo Name: MH006
Embryo stage:ST28 (CONFIRMED BY FABIANA)
Staging description:
Fixation:PF
Cryoprotection: Yes
Material label and storage: Stored in a vial filled with 30% sucrose PBS solution labelled MH006 at room temperature.
Experiment details
Objective: To successfully cut good histological quality ST28 sections
Procedure: Detailed experimental protocol can be found here
Comments: This was not a serial sectioned embryo. The quality of the tissue was very poor. The decision was made not to complete cutting the whole embryo for this reason.
Cryostat Sectioning Cryostat settings (for a more detailed protocol visit Kubke_Lab:Cryomicrotomy)
Cryostat | Histology Lab |
Knife | |
Day Cut | 19/1/11 |
Knife Angle | 0.5 |
Chamber Temp | -17 |
Object Temp | -17 |
Glass Slides | Gelatin Subbed slides |
Plane of section | Coronal |
Number of slides | 12 |
Observations | The poor quality of the sections was evident before staining. Did not complete sectioning due to this. |
(Include in your observations, eg ,were the sections serial, was any section lost, was quality assessed, etc)
Date | 20/1/11 | |
Defatting and rehydration step | ||
Solution | Time | Comments |
steps omitted | ||
Staining and differentiation step | ||
Solution | Time | Comments |
Water | 3 minutes | |
Cresyl Violet | 5 minutes | |
50% alcohol | 1 minute | |
70% alcohol | 1 minute | Additional step added following conversation on 18/1 |
70% alcohol acetic acid | 2 minutes | Additional step added following conversation on18/1 |
95% alcohol | 1 minute | |
100% alcohol 1 | 1 minute | |
100% alcohol 2 | 1 minute | |
Xylene 1 | 4 minutes | |
Xylene 2 | 3 minutes | |
Xylene 3 | 2 minutes | |
Coverslip | 15 minutes | The slides used in this experiment were not frosted therefore a diamond pencil had to be used to label the slides. During coverslipping the liquid made it quite difficult to see the writing. |
Results
Please see following entry To assess these sections the following questions were included:
- What are the cell types present?
- Are the cells intact?
- What improvements can be made ?