- 9:00 am Lab meeting/journal club. Room 502-B51. Check your email to get the papers we are discussing.
- 9:00 AM : Lab meeting
- 11.00 AM: Made an incision at the wind-bud level removing the lower half of the embryo MH006 . Placed the embryo in a larger mould due to previous issues with the eye protruding out of the mould. The embryo was placed in the cryostat( temp -17/-17).
- (Note: what is the embryo number?--MF Kubke 06:02, 20 January 2011 (EST)) ( I have added the embryo name and I will be populating the page today --Malisha Hettiarachchi 15:03, 23 January 2011 (EST).)
- (Note: Any reason why the page is still empty?--MF Kubke 05:06, 3 February 2011 (EST))
- 11.50 AM: The eye was still protruding indicating the mould not being deep enough. Added more OCT and allowed for incubation.
- 12.05PM: Placed mould onto chuck and inserted it into the cryostat. Will allow for approximately 30 minutes incubation time. --Malisha Hettiarachchi 18:13, 18 January 2011 (EST)
- (Note: Thanks for pointing out the issues with the depth of the molds. I will look into it --MF Kubke 18:15, 18 January 2011 (EST))
1.00 PM onwards: Began cutting from the head region towards the wing. Slide 1-2 the temperature fluctuated from -16/-16 to -16/-16. The sections were 20 microns thick. The tissue did not look intact and very discontinouos --Malisha Hettiarachchi 21:33, 18 January 2011 (EST).
- Slide 3: The thickness of the sections was increased from 20 microns to 40 microns , still the tissue looked the same as previously. I increased the " waiting time" before mounting from 45 seconds to a minimum of 1-2 minutes and it improved the sections but it is not very consistent.
- Slide 6-9 : Focused on mounting the sections very slowly and it seemed to be more consistent . The tissue still does have holes.
Plan for tomorrow:
- Nissl stain MH006 sections to determine if the level of subbing is adequate. Ask Felipe if resubbing is an option following protocol A ( cold).
- Find a new box of glazed slides to sub with gelatin.
- Determine if cryoprotection is a factor in the discontinuity of the tissue.
- Attempt to dissect an embryo
--Malisha Hettiarachchi 04:08, 19 January 2011 (EST)
Nissl staining of sections cut yesterday. (RC009)
- 10:30am-12:30pm Stained odd numbered slides.Coverslipped immediately afterwards.
- 1:00pm-2:15pm Stained even numbered slides.Coverslipped immediately afterwards.
Organized the lab also during morning incubation times.
Sections didn't fall of the slides indicating that the subbing was successful
- 2:45pm Working on my online entries. --Reuben Cutfield 20:47, 18 January 2011 (EST)