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  • Real Time Observation of Neutrophil White Blood Cell Recruitment to Bacterial Infection In Vivo

    The zebrafish (Danio rerio) is an emerging vertebrate model organism to study infection. The transparent larva comprises a fully functional innate immune system and enables live imaging of fluorescent immune cells in transgenic animals. Zebrafish infection models have been developed for both the human bacterial pathogen Shigella flexneri and the natural fish bacterial pathogen Mycobacterium marinum. Importantly, whilst S. flexneri causes acute infection and is typically used as an inflammatory paradigm, M. marinum causes a chronic disease similar to tuberculosis in humans. Here, we use real time fluorescence microscopy to image transgenic zebrafish larvae with neutrophils (granulocyte white blood cells) expressing the green fluorescent protein eGFP.
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  • C. Elegans

    Work Efficiently in Developmental Biology with Stereo and Confocal Microscopy: C. elegans

    For scientists, technicians, and teachers working with the worm C. elegans in the research lab or classroom, this report is intended to give useful information to help improve their daly work. The aim is to make the work steps of worm picking, transgenesis, RNA interference, screening, and functional imaging efficient. It also details the various possibilities for equipping a research worm lab or biology classroom/teaching lab explaining worm methods.
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  • Infection of Zebrafish Embryos with Intracellular Bacterial Pathogens

    Transparent zebrafish embryos have proved useful model hosts to visualize and functionally study interactions between innate immune cells and intracellular bacterial pathogens, such as Salmonella typhimurium and Mycobacterium marinum. Micro-injection of bacteria and multi-color fluorescence imaging are essential techniques involved in the application of zebrafish embryo infection models.
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  • Intravenous Microinjections of Zebrafish Larvae to Study Acute Kidney Injury

    We describe a technique of microinjecting the aminoglycoside, gentamicin, into 2 days post-fetilization (dpf) zebrafish larvae to induce acute kidney injury (AKI). We also describe a method for whole mount immunohistochemistry, plastic embedding and sectioning of zebrafish larvae to visualize the AKI mediated damage.
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  • The Principles of Modulation Contrast

    Modulation contrast creates 3D-like images of unstained specimens, rendering it the contrasting method of choice for applications like e.g. in vitro fertilization, where DIC is not possible (due to the usage of plastics) or phase contrast does not deliver satisfying results. This tutorial explains the optical elements in the light path and the operating mode of the modulation contrast.
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  • Integrated Modulation Contrast (IMC)

    Hoffman modulation contrast has established itself as a standard for the observation of unstained, low-contrast biological specimens. The integration of the modulator in the beam path of themodern inverted microscopes allows a wide range of brightfield or phase objectives to be used, rather than a small selection of special objectives.
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