Kubke Lab:/Notebook/Cranial nerve development/2010/11/25

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Cryostat Training, Day 2

9:30am Malisha and Reuben met at Satya's lab. Reuben resumed cutting practice (of embryo RC001) and Malisha began her cutting training (of embryo MH001).Turns were taken cutting the tissue at different thicknesses varying the technique slightly each time in an attempt to learn the properties of the cryostat.

(Note: Please provide the name of the embryo that was processed on this day --MF Kubke 04:23, 20 January 2011 (EST))

Nissl Stain practice

5 slides containing sections from RC001 were put through a Nissl stain procedure by Reuben and Malisha to assess the quality of the sectioning. Alongside the supervisor the stained slides were viewed under a microscope and the tissue was deemed to have been destroyed. A reconsideration of the sectioning technique taught by Satya was discussed. It was decided that the sections should not be re-frozen following sectioning, rather kept at room temperature. Also a thicker section was postulated to be more suitable so that there was more biological tissue present to preserve the cytoarchitecture.

(Note: Please provide the name of the embryo that was stained this day, how the material is labelled and where it is stored--MF Kubke 04:24, 20 January 2011 (EST))- this has been provided in the embryo pages.
  • From this staining exercise we also learnt not to use alcohol based markers and to instead use pencil/diamond pencil. A suggestion by Fabiana was to cut the embryo at a warmer temperature i.e -16 C to -19 C--Malisha Hettiarachchi 02:46, 28 January 2011 (EST).

Reuben also provided me with the Nissl stain protocol shown here (Revision as of 21:37, 20 January 2011)--Malisha Hettiarachchi 02:41, 28 January 2011 (EST)

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