Kubke Lab:/Notebook/Cranial nerve development/2010/12/21

Cranial Nerve Development

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General Entries Met with Reuben regarding progress and reporting process.--MF Kubke 18:12, 20 December 2010 (EST) I will be unavailable on the afternoon, undergoing training with Cornelia and Jacqui on the new microscope --MF Kubke 17:24, 16 December 2010 (EST) Personal Entries Malisha On leave --MF Kubke 17:45, 16 December 2010 (EST) Reuben Following a meeting with Fabiana regarding poor record keeping the entries for the past week were updated from my lab notebook (paper version). I collected all my slides containing the embryonic sections that I cut (excluding embryo RC003 which was staged and cryoprotected by me but cut by Malisha and Fabiana) and put them into labeled slide folders (two folders one labeled RC-001 and the other RC-002&RC-004). (Note: Please make sure you add this information to the individual embryo record pages --MF Kubke 05:16, 20 January 2011 (EST)) 1:00pm Began working on my personal objectives shared document. I am adding a table of all my benchmarks and objectives such that the progress made for each point can be noted, seen and checked off by myself and Fabiana. I am currently completing the table now filling in the progress for each point.--Reuben Cutfield 19:30, 20 December 2010 (EST) 2:30pm I have completed filling out my objectives table for the summer. I wrote comments on my progress for each objective mentioned.I will now be constructing a section in my shared document which will allow me to develop a research question for my project with my progress being observable. --Reuben Cutfield 20:33, 20 December 2010 (EST) 2:45pm Read: Developmental organization of the glossopharyngeal nucleus in the embryonic chick brainstem slice as revealed by optical sectioning recording, by (Sato et al 2002). Added a proposed question to the shared personal objectives document. 3:30pm Read: MOLECULAR PROFILING INDICATES AVIAN BRANCHIOMOTOR NUCLEI INVADE THE HINDBRAIN ALAR PLATE by Ju, Aroca, Luo, Puelles and Redies, 2004. Was helpful in some respects, made a few notes from it. (Note: Please create pages in the journal club area for these papers --MF Kubke 05:16, 20 January 2011 (EST)) 5:00pm Looked at slides 1-5 (RC004) with Fabiana. The sections on slide 5 were the best and were good enough quality for analysis. She has instructed me to go ahead and cut a few whole embryos using the same settings as in slide 5 which were: NB: RC004 was a stage ST25 embryo. 20-25 micron thick sections -19 degrees C OT and CT 1.5 degrees blade angle Notes Cut as quickly as possible through the section (being careful though not to 'hit' the blade with the block, as this will damage *the equipment) Use fixed tissue, PFA (not cryoprotected in 30% sucrose) Allow a delay between cutting and mounting to prevent curling Cut away by hand with a razor blade the side of the OCT block such that the cutting surface makes a diamond shape. This helps prevent curling and also is best for mounting. Keep you hand as steady as possible during mounting. Aim to minimize movement of the slide during mounting. Try to mount as quickly as possible, a 'jump' as opposed to a 'roll' is most effective. Fabiana