Etchevers:Notebook/Genomics of hNCC/2008/04/16

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CHD7 immunohistochemistry

Had to manually paste this model into the page, as for the entry for eye development project. Not true for the preceding test entries?

Will try to enter yesterday's gestures in a post on yesterday's date but with today's timestamp; I didn't have time to enter them yesterday evening.

Today: Rinsed slides in PBT 3x 5', the third bath was actually more like 30'.

Incubated 1h with goat anti-rabbit IgG - Alexa Fluor 488 diluted to 1/200 in PBT in the dark in humid chamber.

Rinsed slides 3x 10' (more or less) in PBT; the first bath was just to get off the coverslips.

Meanwhile, thawed out the Prolong Gold antifade mounting medium in the 37°C bacterial shaker. Mounted slides just before lunch. Did the rounds with nailpolish.

Took photos on the fluorescent microscope on 1st floor. Got interrupted for meeting on future collaboration with use of human kidney tissues in chromatin immunoprecipitation.

  • To take photos in Leica's IM50 software, right-click on "configuration" to choose one for the given exposure. Can save under another name but very helpful to have same exposure conditions for experimental as for no 1° antibody control. Optically it was very dim and might have had better photos on the confocal. Nonetheless the photos were quite nice.
  • Of the little levers on the fluorescent light source tube, the one furthest back is the obturateur.

Submitted an Abreview to Abcam for their CHD7 antibody ab31824; lot 418948.

I'll see if I can post photos in here. I took them first so na!

Embryonic neural tube, human stage C13 at cervical level.

CHD7 protein like the mRNA is expressed in the tube but excluded from the roofplate and floorplate, seemingly expressed though in developing postmitotic motoneurons and migrating neural crest cells (stream on right?). Also in the dermamyotome and ectoderm, not in sclerotome or notochord, heart or yolk sac. I won't post all photos because I wonder if making a lab notebook available with images that I might want to use in a journal article constitutes "previous publication"? Well, there are plenty to go around.

CHD7 DAPI NT2 20x small.jpg

And all this stuff is still in my real physical notebook. Takes much more time so far to enter it on the computer, though I type faster than I write. And I need a physical notebook when doing wet lab work for scribbling things down as I go along or checking off the recipe for a PCR.

  • Alethea 13:52, 16 April 2008 (EDT):