Kubke Lab:Research/CND/Records2010-2011Summer/MH003: Difference between revisions
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=Results= | =Results= | ||
The description of the staining procedure can be found [[Kubke_Lab:/Notebook/Cranial_nerve_development/2010/12/16|here]] | |||
{|border="1" | |||
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|colspan="2"| Date||16/12/2010 | |||
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|colspan ="3" align=center | '''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 || || | |||
|- | |||
|Xylene 3|| || | |||
|- | |||
|Xylene 2 || || | |||
|- | |||
|Xylene 1 || || | |||
|- | |||
|100% alcohol 2||Omitted || | |||
|- | |||
|100% alcohol 1 ||2 min || | |||
|- | |||
|95% alcohol||2 min || | |||
|- | |||
|75% alcohol||Omitted || | |||
|- | |||
|70% alcohol||2 min|| | |||
|- | |||
|Water|| || | |||
|- | |||
|colspan ="3" align=center | '''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|| || | |||
|} | |||
=Images= | =Images= | ||
=Summary= | =Summary= | ||
Revision as of 19:34, 3 February 2011
| 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
Results
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 | ||
| Xylene 3 | ||
| Xylene 2 | ||
| Xylene 1 | ||
| 100% alcohol 2 | Omitted | |
| 100% alcohol 1 | 2 min | |
| 95% alcohol | 2 min | |
| 75% alcohol | Omitted | |
| 70% alcohol | 2 min | |
| Water | ||
| 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 | ||
