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

From OpenWetWare

Jump to: navigation, search
Cranial Nerve Development Main project page
Previous entry      Next entry

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


Personal tools