Digital sight ds l3

Cells were stained with Oil-Red-O (Sigma–Aldrich) as described previously [30 (link)]. 3T3-L1 cells were plated on a 24-well plate and induced to differentiate using the DMI method described earlier. The cells were rinsed with phosphate-buffered saline (PBS), fixed with 4% paraformaldehyde (PFA; Wako) for 14 min, and then stained with 3 mg/mL Oil-Red-O (in 60% isopropanol) for 10 min at room temperature. After staining, cells were washed once with 60% aqueous isopropanol and twice with PBS. After washing, the cells were observed using an ECLIPSE Ti-U inverted microscope (Nikon, Tokyo, Japan). Cell images were captured with a CCD camera (digital sight DS-L3, Nikon). Additionally, after the dye was extracted for 10 min with isopropanol, the absorbance was measured at 490 nm using a DTX 880 Multimode Detector (Beckman Coulter, Brea, CA, USA). All experiments were performed at least three times.

The intestinal tissues were fixed in buffered 10% formalin and then processed for standard (5 μm) paraffin sections. Then, sections were incubated for 30 min in 3% H₂O₂, and nonspecific binding was blocked with normal serum. The PDH was incubated with primary antibodies against PDH overnight at 4 °C. The same concentration of normal serum served as a negative control. The bound antibodies were eventually detected with the biotin-streptavidin peroxidase system. Then, samples were incubated with 3,3′-diaminobenzidine (DAB) peroxidase substrate solution, and the slides was counterstained with haematoxylin. The immunostained images were captured using a digital camera microscope (Nikon Digital Sight DS-L3, Tokyo, Japan).

A densitometer DEN-1 from the company Grant-bio was used to determine the McFarland value. A digital control heated water bath with in-built shaking tray from Thermoline Scientific was used to incubate the bacteria. The Foodsafer Vacuum sealer was used to prepare the sample for exclusion of oxygen. A Nikon digital sight DS-L3 at a 490 nm excitation wavelength and live-dead optical filters were used to take the fluorescence microscopy images. To measure the absorbance at 490 nm an ELx800 absorbance microplate reader from Bio-Tek with the corresponding software Gen5 was used. To measure the absorbance at 540 nm a Synergy HT multi-detection microplate reader from Bio-Tek with the appropriate software KC4 was used.

Cells were seeded in 12-well plates at a concentration of 1 × 10⁵ cells/mL/well. After a 24 h-exposure, culture media were removed gently and replaced with the Live Cell Imaging Solution (Invitrogen, Burlington, Canada). Treated cells were observed with a Nikon TE300 microscope connected to a Nikon Digital Sight DS-L3 camera.

The studied specimens were deposited at the herbarium of the Institute of Microbiology, Beijing Forestry University (BJFC), with color terms following those outlined by Petersen (1996 ). Sections mounted in 5% KOH and 2% phloxine B (C₂₀H₂Br₄C_l4Na₂O₅) were studied at a magnification of up to 1,000 × using a Nikon Eclipse 80i microscope and phase contrast illumination. A Nikon Digital Sight DS-L3 camera was used to photograph microscopic structures. We also used other reagents, including Cotton Blue and Melzer's reagent to observe micromorphology following Wu et al. (2022b (link)). To show the variation in spore sizes, 5% of measurements were excluded from each end of the range and shown in parentheses. At least thirty basidiospores from each specimen were measured. Stalks were excluded from basidia measurements, and the hilar appendage was excluded from basidiospore measurements. The following abbreviations were used: KOH, potassium hydroxide (5%); L, mean length (arithmetic average of all basidiospores length); W, mean width (arithmetic average of all basidiospores width); Q, L/W ratio for each specimen studied; n (a/b), number of basidiospores (a) measured from a given number of specimens (b).