Kubke Lab:Research/CND/Records2010-2011Summer/MH003
| Cranial Nerve Development | Experiment |
Embryo details
Species:Gallus gallus domesticus
Embryo Name:MH003
Embryo stage:20-22
Staging description:Staged by Fabiana unsure of what the criteria was.
Fixation:PF
Cryoprotection:NO
Material label and storage: Stored in a vial labelled MH003 filled with PF at room temperature
Experiment details
Objective:
Procedure:
Cryostat Sectioning
Cryostat settings (for a more detailed protocol visit Kubke_Lab:Cryomicrotomy)
| Cryostat | Histology Lab |
| Knife | |
| Day Cut | |
| Knife Angle | |
| Chamber Temp | |
| Object Temp | |
| Glass Slides | Polysine |
| Plane of section | Coronal |
| Number of slides | 9 |
| Observations | Waiting for approximately 30 seconds or more after cutting a section reduced the curling of OCT and shaping the mould appropriately |
(Include in your observations, eg ,were the sections serial, was any section lost, was quality assessed, etc)
Comments: Slides 1-4 cut by Fabiana, slides 5-9 cut by Malisha
The description of the staining procedure can be found here
| Date | 16/12/2010 | |
| Defatting and rehydration step | ||
| Solution | Time | Comments |
| Water | Omitted | |
| 70% alcohol | 3 min | |
| 75% alcohol | Omitted | |
| 95% alcohol | 3 min | |
| 100% alcohol 1 | 3 min | |
| 100% alcohol 2 | Omitted | |
| Xylene 1 | 15 min | |
| Xylene 2 | Omitted | |
| Xylene 3 | Omitted | |
| Xylene 2 | Omitted | |
| Xylene 1 | Omitted | |
| 100% alcohol 2 | Omitted | |
| 100% alcohol 1 | 2 min | |
| 95% alcohol | 2 min | |
| 75% alcohol | Omitted | |
| 70% alcohol | 2 min | |
| Water | 4 dips | |
| Staining and differentiation step | ||
| Solution | Time | Comments |
| Water | 4 dips | |
| Cresyl Violet | 5 min | |
| 50% alcohol | Omitted | |
| 70% alcohol | 2 min | |
| 70% alcohol acetic acid | Omitted | |
| 95% alcohol | 2 | |
| 100% alcohol 1 | 2 | |
| 100% alcohol 2 | Omitted | |
| Xylene 1 | 2-3 min | |
| Xylene 2 | Omitted | |
| Xylene 3 | Omitted | |
| Coverslip | ||
Results
Slides 1-4: Feedback from Fabiana after looking at these slides was to cut below a temperature of -27C and at 23 microns. Please refer to following entry
