The arterial wall thickness tunica intima and media (IM), tunica media, and tunica intima were measured in transverse histological sections. Measurements were performed in selected fields of one Masson trichrome-stained histological section of each case using a Digital Sight Camera Control Unit (Nikon Digital Sight DS-5M-L1) coupled to a Labophot-2 Nikon microscope. Four different measurements were performed directly in each histological section using digital camera tools. The arterial sections were divided into four-quadrant. Measurements were done in each of the four quadrants, always avoiding damaged tissue if it was present. The median of the four sizes was considered the value of the measure. The tunica media thickness was considered to be the perpendicular stretch between the innermost and outermost elastic lamina. The tunica adventitia was not measured because it was not possible to determine the outer limit of this tunica in all cases.
Digital sight ds 5m l1
Manufactured by Nikon
Sourced in Japan
The Digital Sight DS-5M-L1 is a digital microscope camera designed for laboratory use. It features a 5-megapixel CMOS sensor and supports image capture, display, and storage functions.
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Lab products found in correlation
3 protocols using digital sight ds 5m l1
1
Arterial Wall Thickness Measurement
2
Hepatitis E Virus Tetherin Expression
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In vivo expression levels of BST2 (Tetherin) were studied in HEV associated acute liver failure biopsies (n = 2) retrieved from archives of Department of Pathology, AIIMS, New-Delhi. Normal liver for control (n = 4) was obtained from autopsies performed for non-hepatic diseases (Pancreatitis) or wedge biopsies of liver carried out during other surgical procedures (lino-renal shunt, splenectomy, esophagectomy). Ethical clearance for use of human tissues was obtained from institutional ethical clearance committee (IEC-49/09.12.2015). Immunohistochemistry (IHC) was performed on 4μm thin formalin-fixed paraffin embedded tissue sections post heat induced epitope retrieval (10 mM Citrate buffer, pH 6.0) using 1:600 dilution of mice anti-BST2 (Cat no: ab88523, Abcam, Cambridge, United Kingdom) and 1:50 dilution of in-house monoclonal mice anti-HEV pORF2 primary antibodies as previously described [15 ]. Splenic and lymph node tissues were used as positive controls for BST2 immunostaining. BST2 (Tetherin) and HEV pORF2 staining was done on immediate serial sections to determine co-expression in similar cell populations (Fig 3I–3T ). Images were taken using Nikon ECLIPSE E600 and Digital Sight DS-5M-L1 (Nikon, Minato, Tokyo, Japan).
3
Wholemount Skeletal Staining of Embryos
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Bone and cartilage of embryos were stained in a wholemount preparation according to the protocol reported by Kelly and Bryden (14) , with slight modifications reported by Yamazaki et al. (15) . Briefly, embryos were fixed in 4% paraformaldehyde in phosphate-buffered saline (pH 7.35; PBS) for 2 h at 4°C. After washing out the fixative, the embryos were stained overnight in 0.01% alcian blue solution: 10 mg of alcian blue 8GX (AB, C.I. 74240; Wako Pure Chemical Industries, Ltd., Osaka, Japan) in 80 mL of 95% ethanol plus 20 mL of 99.7% acetic acid. The stained embryos were then rinsed and immediately neutralized overnight with 1% potassium hydroxide (KOH) in 70% ethanol, rehydrated through a graded ethanol series, and stained for 24 h in 0.002% (w/v) alizarin red S (AR, C.I. 58005; Wako) in 0.5% KOH. After rinsing, the specimens were macerated in 2.0% KOH at room temperature and cleared successively in 3:1, 1:1, and 1:3 mixtures of 0.5% KOH and glycerol for 8-24 h each. The stained embryo specimens kept in 50% glycerol (1/1) were examined under a stereomicroscope (SZ61; Olympus Co., Tokyo, Japan) equipped with a camera (Digital SIGHT DS-5M-L1, Nikon Co., Tokyo, Japan).
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