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 ?
