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| Research Focus & Application: |
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At the Laboratory of Molecular Biology (NINDS/National Institutes of Health, P.I.: Ron McKay), we focus on stem cell biology and Neuroscience research. My personal focus is on neural stem cell signal transduction and the effects of their activation in the context of neurological disease. The application here is tangential to my work. I needed to know that the flattened shape of cell aggregates cultured as spheres was a result of pressure from the cover slip and not a limitation of the acquisition process; I thought a chunky dead window bug or some tree moss (few hundred microns thick) would give me the answer.
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| Dr. Andreas Androutsellis-Theotokis |
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| Microscopy and Imaging Methods: |
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The dead bug was placed on a coverslip and mounting medium was used to hold it in place without the need to squeeze the specimen. Autofluorescence was captured using an Axiovert 200 M microscope with an Appotome (L grid with no averaging or filtering) and a 10X Objective (Plan-Apochromat 10x/0.45 M27) with a single channel (GFP) Z-stack collection (with 10 micrometer steps for speed) and an ApotomeCam HRm camera. The image above represents a maximum intensity projection of the Z-series including an x-z projection (top) and y-z projection (right). We use Zeiss because of the system’s ability to take great resolution images with a-speed and efficiency at a lower cost than other comparable systems. | | Dead window bug (at x10 magnification) using an Apotome with maximum intensity projection. |