Kubke Lab:/Notebook/Cranial nerve development/2010/11/26

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General Entries

Cryostat Training Continued

  • 9:30am We began cutting the already molded, frozen and trimmed embryo RC001 with an object temperature of -18 degrees and chamber temperature of -18 degrees also. The mold was then sectioned at 10 microns, 12, 14,16 and 20 microns with a slide-full of sections being mounted for each thickness.
  • 10:30am: The chamber temperature was lowered to -20 degrees and sections were cut and mounted at 12 and 14 microns.
  • 10:45 am: The chamber temperature was lowered to -23 degrees and sections were cut and mounted at 10, 12 and 14 microns
  • 11:00am We then removed the embryo RC001 and prepared the mold for MH001, froze the mold and set it up in the cryostat. The embryo was then trimmed at the temperature the cryostat was at (OT: -18 CT: -23).
(Note: Can you clarify, the entries from the previous day says that the MH001 was cut (Kubke_Lab:/Notebook/Cranial_nerve_development/2010/11/25) and the entries from the prior day says that the embryo was put on a mold then (Kubke_Lab:/Notebook/Cranial_nerve_development/2010/11/24) can you please clarify? --MF Kubke 04:06, 3 February 2011 (EST))
  • 1:00pm We then lowered the temperature of the cryostat to an object temperature of -23 degrees and a chamber temperature of -24 degrees and the cryostat was left to equilibrate for 1hr 30min.
  • 2:30 Sections were cut and mounted by Reuben at 10, 12, 14 and 20 microns.
  • 2:45 The cryostat was then set by Reuben to an object temperature of -14 degrees and a chamber temperature of -16 degrees and the apparatus left to equilibrate for an hour.
  • 3:45 A single slide at 10microns was sectioned and mounted by Reuben.

--Reuben Cutfield 20:02, 30 November 2010 (EST)

Personal Entries

Reuben

  • I completed a bullet-list of my personal objectives I wish to work towards during the summer project. For use in meeting with Fabiana. --Reuben Cutfield 21:15, 30 November 2010 (EST)

Malisha

  • From todays sectioning exercise we learnt that lowering the temperature to -18 C OT/CT worked well. At stage 22 the warmer temperature works . During the experiment however the tissue is quite delicate therefore the temperature shouldn't be changed.
  • Another thing to monitor is the sharpness of the blade . This will be done by marking which area of the blade was used and for how long.--Malisha Hettiarachchi 02:58, 28 January 2011 (EST)


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