Ht7800 tem

Tilting images of unroofing specimens were modulated as performed in a previous study (Morone et al., 2006 (link)). The tilt angles were changed every 2° intervals from −60° to +60° using HT 7800 TEM (Hitachi High-tech, Tokyo, Japan). Corrections of all images and take alignment were performed by EMIP-EX software with HT 7800 TEM (Hitachi High-tech).

First, intestine and lung tissues were fixed with 1% osmic acid in 0.1 M phosphate buffer (PBS, pH 7.4) for 2 h at room temperature in the dark, and the samples were rinsed three times for 15 min each time. Tissues were dehydrated in graded ethanol (30%, 50%, 70%, 80%, 95%, and 100% for 20 min each) and then dehydrated twice in 100% acetone for 15 min each. Pure EMbed 812 resin (SPI, 90529-77-4) was poured into the embedding plate, and samples were placed into the embedding plate and maintained in a 37°C oven overnight. Then 60 to 80-nm thick ultrathin tissue sections were sliced, and the 150-mesh copper mesh was removed. The slices were placed in saturated alcohol solution with 2% uranium acetate for 8 min in a dark room for staining, then rinsed with 70% ethanol and ultrapure water three times. Next, 2.6% lead citrate was added and allowed to stain for 8 min. After washing and drying, the cuprum grids were placed on the grids board and dried overnight at room temperature. For analysis, images were obtained via HT7800 TEM (HITACHI Tokyo, Japan). Images were presented at 12,000× and 20,000× magnification.

NRK-52E cells were collected and fixed in 2.5% glutaraldehyde for 2–4 h. The cells were then immobilized with 1% osmium acid for 2 h, dehydrated using a graded alcohol series, permeated overnight with acetone, and stained with uranium and lead. Finally, the samples were observed and the images were captured under a HT7800 TEM (Hitachi, Japan).