Kubke Lab:Research/CND/Records2010-2011Summer/MH005

From OpenWetWare
(Redirected from Kubke Lab:CND/MH005)
Jump to: navigation, search
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

Images

Summary