Kubke Lab:/Notebook/Cranial nerve development/2011/01/28

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General Entries

  • Please note I added the plane of sectioning field to the cryostat template. Please make sure you include that information in your records.
  • Please make sure you check that your links are working, I am finding several dead end links in your journal entries
  • Please make sure that you do not remove fields from the embryo records template when you create a page

--MF Kubke 00:54, 27 January 2011 (EST)

Personal Entries


  • 9.15 AM: Check and reply to notes and feedback from previous journal entries . Edited entry from 27/1 --Malisha Hettiarachchi 15:40, 27 January 2011 (EST) .
  • 9.40 AM: Prepare Nissl staining solutions . --Malisha Hettiarachchi 15:40, 27 January 2011 (EST)
  • 12.00 PM :Meeting with kubke
  • 1.00 pm onwards: Began to populate embryo pages, MH001,MH002 .--Malisha Hettiarachchi 21:18, 27 January 2011 (EST)
  • Questions to answer:

1. What criteria should be included when assessing the quality of a Nissl stain?

Neurons contain Nissl substance (mainly made up of RER). Nissl substance is quite basophilic therefore will be stained with basic aninline dyes.

Used the following resource to began creating a set of criteria to evaluate the Nissl stained sections of the embryos [1]

2. Why is there a series of dehydration and rehydration steps ? Why is there a "differentiation step" ?
Cresyl violet is an organic compound which stains neurons and glia. The reason for the dehydration and rehydration steps is not to get rid of the OCT as previously assumed by me but instead it is to cause the myelin and other components to lose colour and for the perikarya or soma containing the cell nucleus to retain colour.

3. Why is it better to cut at a warmer temperature?

http://physio.ucsf.edu/rubenstein/protocols/PDFs/Cobos_ISH.pdf , this protocol used by another lab group also cuts at relatively a warm temperature of -16C. From their results it is the optimal temperature for the embryo brain. --Malisha Hettiarachchi 03:11, 28 January 2011 (EST)


  • Will be taking personal day off


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