Macrofire monochrome progressive scan ccd camera

PIZZ and PIMM cells were transfected with or without 22 μg WT SVIP plasmid or siSVIP and were preserved in 1% glutaraldehyde in Tyrode’s buffer, rinsed with Tyrode’s buffer, scraped from the culture dishes, and pelleted in centrifuge tubes. Pelleted cells were rinsed with 2-mercaptoethanol in 0.1M Na cacodylate buffer (2-ME buffer), post-fixed in 1% osmium tetroxide in 2-ME buffer, and washed in 2-ME buffer. The samples were dehydrated in a graded series of ethanols and propylene oxide and embedded in epoxy resin (Taab 812 Resin, Marivac Industries, Montreal, CA). Ultrathin (60–70 nm) sections were counterstained with uranyl acetate and lead citrate and observed using a Hitachi 7600 transmission electron microscope (Hitachi High-Technologies America, Schaumburg, IL) equipped with a Macrofire monochrome progressive scan CCD camera (Optronics, Goleta, CA) and AMTV image capture software (Advanced Microscopy Techniques, Danvers, MA).

Transmission electron microscopy was used to evaluate autophagic vacuoles in the liver tissues of mice. Mouse liver tissues were fixed with a solution containing 2.5% glutaraldehyde and 1% formaldehyde in 100 mM sodium phosphate buffer (pH 7.2) at 4 °C and then postfixed with 2% osmium tetroxide in a 100 mM sodium cacodylate buffer (pH 7.4) for 1 h at 24 °C. The fixed samples were dehydrated in graded alcohol and embedded in epoxy resin (Taab 812 Resin; Marivac Industries, Montreal, Canada). Ultrathin tissue sections (60–70 nm) were stained with uranyl acetate and lead citrate and detected with a transmission electron microscope (Hitachi 7600, Hitachi High-Technologies America, Schaumburg, IL, USA) equipped with a Macrofire monochrome progressive scan CCD camera (Optronics, Goleta, CA, USA) and AMTv image capture software (Advanced Microscopy Techniques, Danvers, MA, USA).

Semi-thin sections were observed with an Olympus BX60 microscope (Tokyo, Japan) between crossed-polarizers. Molecular orientations were visualized with a gypsum first-order red retardation plate inserted at 45° between the polarizer and analyzer [33 (link)]. Images were captured with a Lumenera Infinity 3 digital camera (Ottawa, ON, Canada). Freeze-dried samples were mounted onto aluminum stubs via adherence to double-sided copper tape. All of the samples were then sputter coated with amorphous carbon before analysis with an FEI Nova 430, operated at 15 kV and a spot size of 4.
Ultrathin sections were imaged with a Hitachi 760 TEM (Hitachi High-Technologies America, Schaumburg, IL, USA) equipped with a Macrofire monochrome progressive scan CCD camera (Optronics, Goleta, CA, USA) and AMT image capture software version 600.335p (Advanced Microscopy Techniques, Danvers, MA, USA) at an accelerating voltage of 80 kV in bright field TEM (BF-TEM) mode. To help to visualize the crystallographic orientation of the hydroxyapatite crystals, ultrathin sections were imaged in BF/SAED mode using a JEOL 2010F (Tokyo, Japan) operating at 200 kV. AFM imaging was conducted on an Asylum MFP-3D (Santa Barbara, CA, USA) operating in AC tapping mode with a nominal scan rate of 1 Hz.