Kubke Lab:/Notebook/Cranial nerve development/2010/12/06

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Reuben

Staining Sections

I performed an H&E stain on the sections cut on Friday, these were from the embryo RC002 kept in PFA without cryoprotection

(Note: are you talking about the embryo before you cut it or about the sections?--MF Kubke 04:25, 3 February 2011 (EST)). They did not fall of the slides which would suggest that using subbed slides did help with adhesion. The Haematoxylin staining was fine however the counterstain with Eosin needed to be stronger and so next time I will keep the sections in the Eosin for longer and perhaps not leave them/wash them in the ethanol for as long (maybe 1sec).
(Note: Please provide a name for the embryo and a link to the record page for that embryo.--MF Kubke 04:37, 20 January 2011 (EST))

I will now stain some of the sections Malisha has cut to see if the cryoprotection made a difference.--Reuben Cutfield 18:48, 5 December 2010 (EST)

(Note: Please provide a name for this embryo and a link to the record page for that embryo. If you did not do this (as appears fron entry below), please strike through the text.--MF Kubke 04:37, 20 January 2011 (EST))

I did not stain Malisha's sections and in lieu I will stage and place in sucrose another embryo (RC003) to which I will perform thick cuts (16-20um) and mount using the razor blade onto polylysine-subbed slides. --Reuben Cutfield 19:46, 6 December 2010 (EST).

(Note: Did you do this? If so provide details for the embryo, otherwise strikethrough the entry --MF Kubke 04:37, 20 January 2011 (EST)).

Malisha

10.20 am: MH002 was moved from the freezer to the cryostat and cut the details can be viewed here

(Note: your entry says you used a knife angle of 0. I thought you were using a higher angle at this time. Could you please confirm?--MF Kubke 04:26, 3 February 2011 (EST))


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