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  • Bruno Vellutini 18:00 on 2012/04/26 Permalink
    Tags: confocal,   


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    First try on the confocal with L. ruber. Samples treated cleared with Murray’s clear and stained with Phallacidin and Propidium Iodide. Since I only used the 40x objective I just managed to capture close ups of the body wall and anterior end. I could only do one slide using the control with murray clear in the confocal, so I will still compare with the samples without murray clear.

    Tissues looked quite ok. By checking out in the scope with DIC the epidermis looked normal and the cilia were intact, it seems. Staining in both (cysteine/control) looked the same under fluorescent lamp.

     
  • Bruno Vellutini 18:00 on 2012/02/26 Permalink
    Tags: confocal,   

    Antibody staining for cyphonautes larvae 


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    Execute the protocol for antibody staining with Chema to test in cyphonautes larvae of Membranipora. Chosen antibody is phospho-histone which binds to histone in chromatin revealing dividing cells.

    26/02/2012

    Day one of the standard protocol. Incubating overnight with rabbit primary antibody diluted 1:2500 in PTx+NGS.

    27/02/2012

    Day two, washes and incubation with secondary (rabbit) antibody (orange-red AlexaFluor 594) diluted 1:200 in PTx+NGS.

    28/02/2012

    Day three, regular washes and addition of Phallacidin with 1:1000 DAPI. Staining for 1h and mounting in glycerol.

    Mounted 3 slides. 1 is good, 2 has most embryos and most dirt, 3 has only a few embryos. Checked the slides in the fluorescent lamp: DAPI and Phallacidin look great and strong; Phospho-histone signal is widespread in the whole embryo, but seems to be slightly stronger in the apical region.

     
  • Bruno Vellutini 08:00 on 2012/01/31 Permalink
    Tags: confocal,   

    8th MIC confocal microscopy course at the Dept. of Biomedicine, University of Bergen 


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    Confocal course: 31st of January – 3rd of February 2012. Program: Program and info 8th confocal course 2012

     
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