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Lsm880 fastairy

Manufactured by Zeiss

The LSM880 FastAiry is a confocal laser scanning microscope that utilizes an advanced airyscan detector to provide high-resolution, high-sensitivity imaging. The system is designed to enable fast, flexible, and efficient imaging of a wide range of samples.

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2 protocols using lsm880 fastairy

1

Correlative Light and Electron Microscopy of Liver

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CLEM was performed using a workflow as described by Urwyler et al., (2015) (link). In brief, mice were intravenously injected with 20 μg of CD31-conjugated antibody to stain sinusoid blood vessels. After 5 min, mice were sacrificed and inferior vena cava were cannulated and livers were perfused (4 mL/min) with Antigenfix (Diapath) for 5 min at room temperature. After excision, 2-3 mm slices of livers were further fixed by immersion in Antigenfix for 1 h at 4°C, washed in PBS and sliced making use of Leica Vibratome VT1200S. Slices were first used for Near Infra-Red Branding to indicate a region of interest (ROI), making use of a Zeiss LSM780 with a MaiTai laser (Bishop et al., 2011 (link)). Next, high resolution confocal stacks were obtained with a Zeiss LSM880 FastAiry. The sample was then processed for volume EM by fixation with Karnovsky buffer and en bloc heavy metal staining as described by Steeland et al., (Steeland et al., 2018 (link)). The sample was mounted onto an aluminum pin and trimmed for imaging with a Zeiss Merlin with 3View2 (Gatan). Overlays of confocal and volume EM datasets was performed based on blood vessel staining, using the ec-CLEM plugin of ICY in order to identify Kupffer cells or monocytes (Paul-Gilloteaux et al., 2017 (link)). The EM datasets were segmented in MIB (Belevich et al., 2016 (link)) and 3D rendering was done in iMaris.
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2

Zebrafish Embryo Microscopy Techniques

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Embryos were mounted in 35 mm glass bottom dishes using 0.7% low melting point agarose. Confocal z-stacks were acquired on a LSM 710 Meta BiG (Zeiss) and a LSM 880 FastAiry (Zeiss) using a LD C-Apochromat W/40×1.1 NA objective. Timelapse imaging was performed on a Dragonfly Spinning Disc microscope (Andor) using Apo λ LWD W/40×1.15 NA. Bright-field images of the whole-mount zebrafish embryos were taken on a Nikon SMZ1270i with Tucsen Michrome 6 camera. Movies of blood flow (Movies 4-7) were recorded with a Prime BSI Express camera at 100 frames per second and captured using a EC-Plan Neofluar 20×0.8 NA objective on an Axio Observer 7 inverted microscope (Zeiss).
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