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Imaging early embryos and stem cells Centre for Trophoblast Research Anne McLaren Laboratory for Regenerative Medicine University of Cambridge Kathy Niakan.

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Presentation on theme: "Imaging early embryos and stem cells Centre for Trophoblast Research Anne McLaren Laboratory for Regenerative Medicine University of Cambridge Kathy Niakan."— Presentation transcript:

1 Imaging early embryos and stem cells Centre for Trophoblast Research Anne McLaren Laboratory for Regenerative Medicine University of Cambridge Kathy Niakan

2 Imaging early embryos Pre-implantation embryos at specific time points Fixed and immunofluorescently labeled secondary antibody emission spectrum: 350, 488, 594, and 650 Zeiss confocal microscope: 20 and 40x oil immersion objectives z-stack images: 15-20 z-sections at 3-5  m (100  m total thickness)

3 Technological requirements for live-cell lineage tracing Microscope: -Fluorescence: epifluorescence, structured illumination, or confocal -Structure of sample: Brightfield, phase-contrast, or DIC CO 2 and temperature control: 5% and 37 o C z-stack images: 50-100  m-thick samples Camera: speed and resolution Software for analyze images: 3D rendering, live-cell lineage tracing Computer: each z-stack is 70MB (8-16GB for each time-course)

4 Current bottlenecks in imaging early embryos and embryonic stem cells Substrate to grow cells: plastic versus glass Imaging 3D structures: loss of resolution Storing and analyzing large data sets Integrating a variety of biological information: quantitative and behavioral analysis

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