Dp70 digital camera system

Small fragments of eWAT was fixed in 4% buffered formaldehyde, embedded in paraffin, and sectioned at 10 μm. H&E images were used for the determination of mean adipocyte size and macrophage count. The sections were observed under an Olympus BX51 attached DP70 Digital Camera System (Tokyo, Japan); the fields were evaluated with final magnification of 20x (50 μm). Digital photographs were taken from each section, adipocyte size expression were quantified using the “ImageJ” image processing software (NIH, Bethesda, MD, USA), and the macrophage count was performed on the average of 4 fields per animal (macrophage/field) [26 (link)–28 (link)].

Histological analysis was performed to detect the impact of MjShoc2 knockdown on tissue morphology. Shrimp were infected with 10⁵ CFU of V. anguillarum. The hepatopancreas were collected and fixed using Davidson’s AFA fixative (30% ethanol, 22% formalin, and 11.5% acetic acid). After fixing for 24 h, the tissue was dehydrated, embedded in paraffin, sectioned at 7-μm thickness and stained with hematoxylin and eosin (H&E). The slides were observed under the BX51 inverted fluorescence microscope (Olympus, Tokyo, Japan) and the images were captured by the DP70 digital camera system (Olympus).

To assess immunoreactive staining for GIP, GLP-1, GLP-2 and PYY, sections were dewaxed in histoclear for 30 mins before being rehydrated with decreasing concentrations of ethanol. Following this, antigen retrieval and cell permeabilization was achieved using a citrate buffer (pH 6.0) at 96°C for 20 mins followed by a cooling period. The sections were blocked with 2.5% bovine serum albumin (BSA) and then incubated with a primary antibody (Table 1) for the respective peptide overnight. Notably, the GLP-1 antibody employed for our studies will only detect truncated GLP-1 peptides and not related GLP-1 proglucagon gene products such as GLP-1(1–37) [9 (link)]. On day 2, the sections were rinsed in phosphate-buffered saline (PBS) twice and incubated with secondary antibody (Alexa Fluor^® 488 for green; Table 1) for 1 hour at 37°C. After two further PBS washes, sections were incubated with DAPI for 15 mins at 37°C [16 (link)]. Finally, the sections were mounted using antifade and coverslips. Stained sections were viewed at 40x magnification using an Olympus IX51 inverted microscope and photographed using a DP70 digital camera system.

Specimens
obtained using punch biopsy (6 mm) were taken from the triangular
fossa and adjacent area before and after decellularization. The specimens
were then exposed to different staining methods to evaluate the changes
in the microanatomy of the auricles after being decellularized using
the two different protocols. All stained samples were visualized using
a CKX41 inverted microscope equipped with a DP70 digital camera system
(Olympus).

To record time-lapsed images of transportation of EGFP-tagged HMGCS1 in live cells, KATO III cells were inoculated on a 3.5-cm glass-bottom plate (Mettek, Ashland, MA, USA) and then transfected with either EGFP-tagged HMGCS1-expressing plasmid (pEGFP-HMGCS1) or EGFP-only expressing vector (pEGFP-C1). Both phase-contrast and fluorescence images of cells were acquired by using an inverted microscope (IX 71, Olympus, Tokyo, Japan) equipped with an Olympus DP70 digital camera system. EGFP was excited by 460–490 nm wavelength, filtered from a mercury light source. The fluorescent emission of EGFP was collected by a 100× oil immersion objective with a numerical aperture (N.A.) value of 1.40 (Olympus, Japan), and filtered by a WIBA filter cube (515–550 nm). The phase-contrast images were acquired prior to fluorescence imaging for displaying the shapes of cells during the recording period. All images were recorded with DP-BSW software (Olympus, Tokyo, Japan). Images of 10 individual cells were acquired at the indicated time points. To elucidate the alteration of the fluorescence in both the cytoplasm and nuclei, the ratio of the fluorescence intensity in nuclear and cytoplasmic areas was determined using ImageJ software (version 1.49, University of Wisconsin, Madison, WI, USA).

H292 cells pre-treated with O-acetyl RT at non-toxic concentration (0, 0.01 and 0.05 μM) for 24 h were seeded in 24 well plates at a density of 100 cells/well and incubated for 7 days. The colony was fixed with methanol at 4 °C for 15 min and stained with 0.1% crystal violet for 15 min. Colony formation was assessed using a phase-contrast microscope (Olympus IX5, Tokyo, Japan) equipped with a DP70 digital camera system (Olympus, Tokyo, Japan).