Kubke Lab:Research/CND/Records2010-2011Summer/MH002: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Line 42: | Line 42: | ||
*H&E stained sections. | *H&E stained sections. | ||
* Fabiana's feedback: Large holes in the sections which are not expected. Cannot determine if the issue is with staining or sectioning --[[User:Malisha Hettiarachchi|Malisha Hettiarachchi]] 19:27, 31 January 2011 (EST). | * Fabiana's feedback: Large holes in the sections which are not expected. Cannot determine if the issue is with staining or sectioning --[[User:Malisha Hettiarachchi|Malisha Hettiarachchi]] 19:27, 31 January 2011 (EST). | ||
{{Note|You need to provide the staining details --[[User:M Fabiana Kubke|MF Kubke]] 04:29, 3 February 2011 (EST)}} | |||
=Images= | =Images= |
Revision as of 02:29, 3 February 2011
Cranial Nerve Development | Experiment |
Embryo details
Species: Gallus gallus domesticus
Embryo Name: MH002
Embryo stage: ST28 (confirmed by Fabiana)
Staging description:
Fixation: PF
Cryoprotection: Yes
Material label and storage: Stored in a vial containing PBS in 30% Sucrose solution overnight at room temperature ON THE 2/12. On the 3/12 the embryo was stored in the histology lab freezer.
- (Note: can you please provide information as to where the slides are stored/labelled?--MF Kubke 04:28, 3 February 2011 (EST))
Experiment details
Objective: To determine what are the best parameters to cut a stage 28 embryo.
Procedure:
- (Note: can you please confirm that the knife angle was 0? I thought we didnt start using that knife angle until later--MF Kubke 04:27, 3 February 2011 (EST))
Cryostat Sectioning Cryostat settings (for a more detailed protocol visit Kubke_Lab:Cryomicrotomy)
Cryostat | Histology Lab |
Knife | |
Day Cut | 3/12/2010 |
Knife Angle | 0 |
Chamber Temp | -23C |
Object Temp | -23c |
Glass Slides | Polysine slides |
Plane of section | Coronal |
Number of slides | |
Observations | Due to the humidity the sections would immediately begin curling when on the metal platform. A razor blade was used to try catch the section therefore not allowing it to curl. This was later disused as a technique as it may have caused additional tissue damage. |
(Include in your observations, eg ,were the sections serial, was any section lost, was quality assessed, etc)
Comments:
Incubated for 30 minutes at a temperature of -23 C OT/CT
Results
- H&E stained sections.
- Fabiana's feedback: Large holes in the sections which are not expected. Cannot determine if the issue is with staining or sectioning --Malisha Hettiarachchi 19:27, 31 January 2011 (EST).
- (Note: You need to provide the staining details --MF Kubke 04:29, 3 February 2011 (EST))