|Cranial Nerve Development||Experiment|
Species: Gallus gallus domesticus
Embryo Name: RC001
Embryo stage: ST22 Confirmed by Fabiana. (see Notebook Entry)
Staging description: Staged as a group exercise by Fabiana, Malisha and Reuben using the Hamburger and Hamilton (1951) staging system.
Cryoprotection: Cryoprotected in 30% Sucrose in 0.1M PBS
Material label and storage: Labeled as RC-001 and stored in sucrose in a vial prior to being cut. The slides produced by RC001 are in a slide folder in the lab labeled RC001.
- Embryo taken from 'stationery draw' where it was stored in PFA.
- As a group we staged the embryo and Fabiana labeled it as ST22, RC001.
- It was put into a vial filled with 30% sucrose, 0.1M PBS.
24/11/2010: Histology Training with Satya see Notebook entry
- The embryo was placed flat on a plastic mould and OCT was then added.
- The embryo was oriented 90° to the side of the block so that coronal sections could be obtained.
- The protruding OCT was cut from the sides of the block.
- The block was mounted onto the chuck at 90° to the chuck.
- A mark was made on the block corresponding to the line drawn on the metallic chuck holder, so that the block could be re-aligned if it had to be moved from the metallic chuck holder.
- The embryo was then sectioned in the cryostat.
Cryostat Sectioning Cryostat settings (for a more detailed protocol visit Kubke_Lab:Cryomicrotomy)
|Cryostat||Leica – CM3050S|
|Knife||MX35 Premier +, 34 degrees, 80mm Thermo Scientific|
|Day Cut||24th November 2010, 10:00am|
|Chamber Temp||The temperature was changed throughout the experiment as part of the experiment. See each individual slide for information on the temperatures used.|
|Object Temp||The temperature was changed throughout the experiment as part of the experiment. See each individual slide for information on the temperatures used.|
|Glass Slides||Permafrost Original Menzel-Glaser Cut Edges, Frosted Ends|
|Plane of section|
|Number of slides||8|
(Include in your observations, eg ,were the sections serial, was any section lost, was quality assessed, etc)
Comments: Tissue from this embryo will not be used for histological analysis. RC001 was used for practice purposes only, it was not stained or coverslipped. The settings of the cryostat and the cutting techniques were altered to learn the effects the settings had on the quality and efficiency of the sections produced. No further work will be done with RC001.