Kubke Lab:Research/CND/Records2010-2011Summer/MH005
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| Cranial Nerve Development | Experiment |
Embryo details
Species: Gallus gallus domesticus
Embryo Name: MH005
Embryo stage: ST28 - ST29
Staging description:
Fixation:
Cryoprotection: No
Material label and storage:
Experiment details
Objective: To determine what are the best parameters to cut a stage 28 embryo.
Procedure:
Cryostat Sectioning Cryostat settings (for a more detailed protocol visit Kubke_Lab:Cryomicrotomy)
| Cryostat | Histology |
| Knife | |
| Day Cut | 14/1/2011 |
| Knife Angle | 0.3 and 0 |
| Chamber Temp | -17C |
| Object Temp | -17C |
| Glass Slides | Superfrost plus slides |
| Plane of section | Coronal |
| Number of slides | 12 |
| Observations | I found that when i waited for a minimum of 1-2 minutes little or no curling was present. When cutting quite fast the section would curl immediately. |
(Include in your observations, eg ,were the sections serial, was any section lost, was quality assessed, etc)
Cresyl Violet staining
For more informattion see Kubke_Lab:Nissl_Stain_Protocol
| Date | ||
| Defatting and rehydration step | ||
| Solution | Time | Comments |
| Water | ||
| 75% alcohol | ||
| 95% alcohol | ||
| 100% alcohol 1 | ||
| 100% alcohol 2 | ||
| Xylene 1 | ||
| Xylene 2 | ||
| Xylene 3 | ||
| Xylene 2 | ||
| Xylene 1 | ||
| 100% alcohol 2 | ||
| 100% alcohol 1 | ||
| 95% alcohol | ||
| 75% alcohol | ||
| Water | ||
| Staining and differentiation step | ||
| Solution | Time | Comments |
| Water | ||
| Cresyl Violet | ||
| 50% alcohol | ||
| 70% alcohol acetic acid | ||
| 95% alcohol | ||
| 100% alcohol 1 | ||
| 100% alcohol 2 | ||
| Xylene 1 | ||
| Xylene 2 | ||
| Xylene 3 | ||
| Coverslip | ||
Comments:
Results
see the following entry
