Ds u3 camera

The tissues of the liver samples were fixed in 10% neutral-buffered formalin for at least 24 h. Then, the tissues were embedded in paraffin and stained with hematoxylin and eosin (H&E) to assess the histologic features of steatosis and inflammation. Frozen liver tissues embedded in an optimal cutting temperature compound (Sakura, Torrance, CA, USA) were cut and stained with Oil Red O. Based on the observation using a laser-scanning confocal microscopy, the images were captured using a Nikon DSU3 camera (Nikon Corporation, Chiyoda Ward, Tokyo, Japan). For the immunofluorescence investigation of intestinal tight junction proteins, frozen intestinal sections were treated with anti-occludin and anti-claudin-2 antibodies (Thermo Fisher Scientific, Waltham, MA, USA), followed by labeling with a FITC-conjugated secondary antibody (Servicebio, Wuhan, China).

Zebrafish were incubated in a culture medium containing 2 µg/mL acridine orange (AO) (Sigma-Aldrich, St. Louis, MO, USA) for 30 min. Before observation, embryos were washed in fresh ERS three times and anesthetized by 0.08% MS-222. Images were captured using a microscope equipped with a Nikon DS-U3 camera (Nikon Inc., Melville, NY, USA). NIS-Elements software (Nikon Inc., Melville, NY, USA) was used.

Cells were collected at different time points and immobilized on 1% 1 X PBS agarose padded slides and were subjected to phase contrast microscopy using a Nikon Eclipse Ts2R microscope (Nikon, Japan) attached with a Nikon DS-Fi3 camera equipped with Nikon Plan Fluor 100X oil Ph3 objective. Time-lapse imaging of live cells harvested at mid-exponential phase (OD₆₀₀ ∼ 0.4) was performed on LB 0.7% agarose padded slides supplemented with or without 0.4% L-arabinose using a Nikon Eclipse Ti microscope (Nikon, Japan) with Nikon DS-U3 camera and Plan Apo 100 X oil objective. Image processing was performed with ImageJ [37 (link)] and Adobe Photoshop CS6 (Adobe Inc. U.S.A).

Pyramidal cells from L5 PL were identified using The Mouse Brain in Stereotaxic Coordinates (4th Edition, Paxinos, and Franklin, 2012) and the Allen Brain Institute’s Mouse Brain Atlas (http://mouse.brain-map.org). The L5 PL neurons were approximately 1.7 mm ventral from the dorsal surface and the cell bodies were 500–700 μm from the medial surface. Individual neurons in these regions were viewed using a 100× oil objective on a Nikon Eclipse Ci-L microscope. Images of the basilar dendrites were acquired using Z-stack projection photomicrographs (0.1–0.9 μm steps) taken using a Nikon DS-U3 camera mounted on the microscope and were analyzed using NIS-Elements D 4.40.00 software. Three to four neurons (middle 80%) were sampled per mouse, and six segments in the same field of view were assessed per neuron (20–50 μm). Each dendrite segment was ~ 1 μm thick and was taken from a 2° or 3° order dendrite. Spine density was expressed as the number of spines/10 μm. To determine the type of dendritic spine, we used parameters described by Risher et al.^{86 (link)}: filopodia, length > 2 µM; long thin, length < 2 µM; thin, length < 1 µM, stubby, width ratio < 1 µM, mushroom, width > 0.06 µM; bifurcated, two or more heads.