Kubke Lab:Research/CND/Records2010-2011Summer/MH006

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Cranial Nerve Development Experiment

Contents

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 Cut19/1/11
Knife Angle0.5
Chamber Temp-17
Object Temp-17
Glass SlidesGelatin Subbed slides
Plane of sectionCoronal
Number of slides12
ObservationsThe 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)




Date20/1/11
Defatting and rehydration step
SolutionTimeComments
steps omitted
Staining and differentiation step
SolutionTimeComments
Water 3 minutes
Cresyl Violet 5 minutes
50% alcohol1 minute
70% alcohol1 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 21 minute
Xylene 1 4 minutes
Xylene 2 3 minutes
Xylene 32 minutes
Coverslip15 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 ?

Images

Summary

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