Kubke Lab:Research/CND/Records2010-2011Summer/MH002
| 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.
The slides are labelled MH002 and stored in a brown folder in Fabiana's lab.
- (Note: can you please provide information as to where the slides are stored/labelled?--MF Kubke 04:28, 3 February 2011 (EST))- Done
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))(Satya had left it on 0 as a "default" knife angle. It was moved just above 0 by you (approximately 0.5). I will check my notes to be sure .)--Malisha Hettiarachchi 16:02, 3 February 2011 (EST)
- (Note: Can you please confirm this? It was my understanding you guys were cutting with a knife angle of about 15 degrees. I remember lowering the angle mid December - Please consult with Satya what her default angle is --MF Kubke 17:04, 3 February 2011 (EST))
Cryostat Sectioning Cryostat settings (for a more detailed protocol visit Kubke_Lab:Cryomicrotomy)
| Cryostat | Histology Lab |
| Knife | 15 |
| Day Cut | 3/12/2010 |
| Knife Angle | |
| Chamber Temp | -23C |
| Object Temp | -23c |
| Glass Slides | Polysine slides |
| Plane of section | Coronal |
| Number of slides | 8 |
| 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 the mould in the cryostat for 30 minutes at a temperature of -23 C OT/CT before placing it on the chuck
- (Note: what did you incubate? I dont understand this entry--MF Kubke 04:35, 3 February 2011 (EST))
Results
- H&E stained sections following this procedure.
- Fabiana's feedback: Large holes in the sections which are not expected. Cannot determine if the issue is with staining or sectioning. ( Corresponding to the following entry) --Malisha Hettiarachchi 19:27, 31 January 2011 (EST).
- (Note: You need to provide the staining details --MF Kubke 04:29, 3 February 2011 (EST))
