Updates from September, 2013 Toggle Comment Threads | Keyboard Shortcuts

  • Bruno Vellutini 23:55 on 2013/09/30 Permalink
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    • Washed Novocrania embryos.
    • Took pictures of Terebratalia in situs: fgfr, pax2/5/8, fz2, fz4. fz10 still needs to be done.
     
  • Bruno Vellutini 17:30 on 2013/09/27 Permalink
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    Fixed 72h 14 °C Membranipora cyphonautes for immuno and in situ.

     
  • Bruno Vellutini 12:40 on 2013/09/26 Permalink
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    Fixed 48h 14 °C Membranipora cyphonautes for immuno and in situ.

     
  • Bruno Vellutini 20:42 on 2013/09/25 Permalink
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    Fixed 28h Membranipora raised at 14 °C for immuno and in situ.

     
  • Bruno Vellutini 14:13 on 2013/09/25 Permalink
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    Fixed 24h Membranipora raised at 14 °C. So they were advanced gastrula and some early larvae.

     
  • Bruno Vellutini 18:36 on 2013/09/24 Permalink
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    Novocrania in situ en, wnt1, wnt5 


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    Started an in situ with Novocrania! I picked 3 specimens of each stage making 12 embryos in total. The stages are: radial, bilateral gastrula, early larva and larva.

    Nano en Nano wnt1 Nano wnt5

    All 12 embryos per well survived the first day.

    25/09/13

    Since engrailed probe had low concentration I diluted what I had (13 ng/µL) into 200 µL of hybe (resulting in 0.5 ng/µL) and added anyway. I’m synthesizing more so that I can add tomorrow.

    26/09/13

    Changed engrailed hybe with nice new probe. However, embryos are really transparent. Larval morphology also does not look normal. I think something is wrong.

    27/09/13

    Engrailed well has not any normal looking embryos anymore. They are all falling apart. In the other 2 wells there are still 4-5 embryos that look ok, but fragile.

    I continued with the washes anyway and incubated with the antibody overnight at 4 °C.

     
  • Bruno Vellutini 17:44 on 2013/09/24 Permalink
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    MAPK inhibitor early stages 


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    Used a 6-well plate with 10 µM U0126 to fix early embryos for immuno staining. Spawning activated at 12h and incubated at 14 °C.

    Embryos fixed after 8h.

     
  • Bruno Vellutini 17:32 on 2013/09/24 Permalink
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    Warning: preg_match_all(): Compilation failed: invalid range in character class at offset 7 in /home/customer/www/phd.organelas.com/public_html/wp-content/themes/p2/inc/mentions.php on line 77

    Tried spawning of Membranipora collected last Thursday and I got thousands of embryos. Will fix some more.

     
  • Bruno Vellutini 17:30 on 2013/09/24 Permalink
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    4D Membranipora Azakenpaullone 5 µM 


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    Folder: BCV_Mm_2013_09_24

    Started a recording with Membranipora with 5 µM Azakenpaullone. Relatively vegetal view.

    Around 18h the embryo detached and was shaking every slice… impossible to track. Kept it overnight just in case.

    25/09/13

    Stopped, embryo was gastrulating.

     
  • Bruno Vellutini 19:59 on 2013/09/22 Permalink
    Tags: EdU,   

    Novocrania EdU series 


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    Set EdU 1:100 for 10 min in 36h embryos. Fixed at 36 and 48h (12h chase).

    23/09/13

    Set another batch of 48h Novocrania embryos (15 per stage) with 10 min EdU. Chased and fixed after 0, 3, 6, and 12h.

    Setting yet another batch of 60h bilateral gastrula for 0 and 12h (overnight).

     
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