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Movie 1.

Making serial sections by histo Jumbo diamond knife. Serial sections are made using a history Jumbo diamond knife (DiATOME, Switzerland) equipped with drainage tubing and collected on a silicon wafer piece (26 mm × 70 mm). Movie 1 shows the entire processes of sectioning and collection of 300 serial ultrathin sections. The procedure for making 300 serial sections is briefly described below. Step 1. Making the first 25 serial sections. The first 25 serial sections are made at 90 nm thickness in ribbon form. After the 25th section is made, the thickness setting is quickly changed to 10 nm. At this setting, the 26th section is not made, but the ribbon is released from the knife edge because the sample block slightly touches the knife edge. Immediately after the ribbon is released, the thickness setting is reset to 90 nm. Step 2. Making up to 100 serial sections. Step 1 is repeated three times to obtain 100 serial sections (four ribbons). When 100 serial sections have been made, the ultramicrotome is paused, and four ribbons are rotated 90°. Then, a polyethylene tape is horizontally placed on the surface of the water and the serial sections are brought to the front space of the knife boat (see also Fig. S7). The polyethylene tape (60 mm × 3 mm) is made from a 0.04-mm polyethylene sheet. Step 3. Making the following 200 serial sections. The sample block is retracted about 500 nm, and then Steps 1, 2 are repeated twice to obtain 300 serial sections (12 ribbons). Step 4. Drainage of boat part. When 300 serial sections have been made, the ultramicrotome is paused, and the valve of the drainage tubing is opened to drain the water, which takes 15-20 minutes. If the drainage speed is too fast, micro-wrinkles will form when the sections stick to the silicon wafer. After the drainage, the silicon wafer is removed. Any water droplets on the wafer should be blown off using a blower.

Ultrastructural analysis of whole glomeruli using array tomography

Takayuki Miyaki, Nozomi Homma, Yuto Kawasaki, Mami Kishi, Junji Yamaguchi, Soichiro Kakuta, Tomoko Shindo, Makoto Sugiura, Juan Alejandro Oliva Trejo, Hisako Kaneda, Takuya Omotehara, Masaki Takechi, Takako Negishi-Koga, Muneaki Ishijima, Kazushi Aoto, Sachiko Iseki, Kosuke Kitamura, Satoru Muto, Mao Amagasa, Shiori Hotchi, Kanako Ogura, Shinsuke Shibata, Tatsuo Sakai, Yusuke Suzuki, and Koichiro Ichimura

J Cell Sci 2024. 137:None-None; doi: 10.1242/jcs.262154