Kubke Lab:/Notebook/Cranial nerve development/2010/12/08

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Cranial Nerve Development
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=General Entries= =Personal Entries=

Reuben
Following a look at Malisha's sections we retrieved some H&E notes from Felipe. I staged an embryo at stage 21 and labeled it as RC003. Then I placed it in 30% sucrose and am waiting for it to sink. Meanwhile I set up a document called 'A brief look at neuroanatomy' and invited Malisha and Fabiana to be contributors for the document. I then put some work into the document to get it started. --Reuben Cutfield 18:02, 7 December 2010 (EST)

The Haematoxylin is purchased as a pure compound and then mixed using the Gills 2 protocol.
 * Emailed Satya asking about our Haematoxylin type - Is Gills 2 --Reuben Cutfield 18:27, 7 December 2010 (EST)

2:00pm I performed 4 separate H&E stains on the sections from embryo RC002 (A-E) and embryo MH002(F). I varied the length of time the slides were sitting in either the Haematoxylin or Eosin and recorded the times spent in each. The slides have been coverslipped and are ready for analysis.

Tomorrow Fabiana will do some cutting and staining to attempt to achieve some satisfactory results. --Reuben Cutfield 20:14, 7 December 2010 (EST)

Malisha
Used an H& E stain to assess the history of the cryostat sections cut on Monday and Tuesday.

Feedback from fabiana : Large holes in the sections which are not expected with a stage 28 embryo (MH002). At this point we cannot determine if it is an issue with the staining or the cut. More reading was done to " tweek" the staining procedure specifically for embryonic tissue--Malisha Hettiarachchi 20:30, 12 December 2010 (EST).
 * At this point the effect of freezing and using the tissue was discussed. MH002 had been frozen and used therefore the tissue damage could be mainly because of this.


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