Eclipse ni u light microscope

Fixed zebrafish larvae were dehydrated in 30% sucrose at 4°C for 3 days, embedded in optimal cutting tissue (OCT) compound (Leica, Germany), and sliced into 14 μM sections. Cryosections were washed with water to remove OCT, incubated in 100% 1,2-propylene glycol for 5 min, and stained with 0.7% Oil Red O at 60°C for 10 min in the dark. Excess dye was rinsed off with 85% 1,2-propylene glycol and PBS to keep the background clean. Sections were imaged using a Nikon Eclipse Ni-U light microscope (Nikon, Tokyo, Japan).
The Oil Red O staining procedure for cells was the same as that of cryosections. The fixed cells were washed with PBS, incubated with 100% 1,2-propylene glycol, stained with 0.7% Oil Red O, and decontaminated in 85% 1,2-propylene glycol and PBS. The staining results were imaged using a Nikon Eclipse Ni-U optical microscope (Nikon, Tokyo, Japan).

Histological evaluation: 6 h (n = 4), 24 h (n = 7), 72 h (n = 8), and 6 days (n = 5) after surgery, the animals under deep anesthesia were perfused with a 0.9% NaCl solution and then with a 10% solution buffered formalin. The brain was removed and resuspended in 10% formalin for 48 h and embedded in paraffin according to the standard technique. Frontal brain sections 5–6 µm thick were prepared at the levels of striatum, sensorimotor cortex, and hippocampus (Bregma 1.1 ± 0.5 mm, Bregma −1.1 ± 0.5 mm, Bregma −4.3 ± 0.5 mm, respectively, according to the atlas by Paxinos G. and Watson C., 1986). Histological examination of brain tissue to identify areas of ischemic damage was performed on hematoxylin–eosin stained sections. Images of brain sections were obtained using a ScanScope CS digital scanner (Leica Biosystems, Vista, CA, USA).
The number of viable pyramidal neurons in the CA1 and CA3–4 fields of the left hemisphere hippocampus were counted using 40× objective (Nikon Eclipse Ni-U light microscope, Nikon Corp., Tokyo, Japan) without the examiner knowing the experimental protocol, using a computer-associated image analyzer (NIS-Elements BR software, Nikon Corp., Tokyo, Japan). The cells with a well-defined ellipsoidal or circular nucleus and a clearly distinguishable nucleolus located in the center of the nucleus were categorized as normal viable neurons [15 (link)].

After being fixed in 4% paraformaldehyde at 4°C overnight, zebrafish larvae were embedded in paraffin and cut into 4 μM sections. Specimens were dewaxed, rehydrated, stained with H&E, dehydrated, cleared, and sealed for routine histology. Finally, H&E sections were captured by a Nikon Eclipse Ni-U light microscope (Nikon, Tokyo, Japan).