Posted by
Mark Cannell-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/imaging-zebrafish-tp7582876p7582878.html
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What objective lens are you using? What is the refractive index of the mountant? If there is a mismatch the signal can drop quite quickly due to spherical aberration. Is the embryo mounted close to the coverslip?
Cheers
On 11/11/2014, at 7:58 am, megha kumar <
[hidden email]> wrote:
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> Hello everyone!
> I am imaging the 256 and 512 cell stage of zebrafish using confocal (Leica
> TCS SP5 II). The embryos are stained with alpha and gamma tubulin antibody.
> I take z stacks and reconstruct the entire embryo using a 3D software. I am
> unable to visualize the deeper cells below although I catch the first top
> most layer pretty well, which gives me a hollow half dome shape in 3D. Any
> ideas why I dont see the deeper cells at 512 cell stage? Is this a
> microscope issue or penetration is not deep enough or something else?
>
> --
> Megha Kumar, Ph.D.
> Young Investigator
> Regional Center for Biotechnology
> 180 Udyog Vihar phase I
> Gurgaon 122016
> India
>
> ph: 8826422770
Mark B. Cannell Ph.D. FRSNZ
Professor of Cardiac Cell Biology
School of Physiology & Pharmacology
Medical Sciences Building
University of Bristol
Bristol
BS8 1TD UK
[hidden email]