|Cranial Nerve Development||Experiment|
Species:Gallus gallus domesticus
Embryo Name: MH006
Embryo stage:ST28 (CONFIRMED BY FABIANA)
Material label and storage: Stored in a vial filled with 30% sucrose PBS solution labelled MH006 at room temperature.
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)
|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)
|Defatting and rehydration step|
|Staining and differentiation step|
|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.|
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 ?