Megaview 2 ccd camera

The presence of Autophagosome in the PR was also observed by TEM. The PR zone of stressed Arabidopsis plants were excised under trinocular stereo microscope (SZ61- OLYMPUS) using sharp razor blade and fixed in 5% Glutaraldeyde in 0.1 M Cacodylate buffer (pH 7.4) 3 hours at room and transferred to 4 °C for continuation of fixation overnight. The samples were further processed according to Galsurker et al.^{55 (link)}. The final sections were placed on grids and sequentially stained with uranyl acetate and Lead citrate for 10 minutes each and viewed with Tecnai 12 TEM 100 kV (Phillips, Eindhoven, the Netherlands) equipped with MegaView II CCD camera and Analysis version −3.0 software (SoftImaging System GmbH, Münstar, Germany).

Diaphragms were removed with the surrounding ribs to prevent folding, washed twice with PBS and were fixed with 2.5% glutaraldehyde and 2% formaldehyde in 0.1 M cacodylate buffer (containing 1% OsO4 and 1.5% potassium ferrycyanide) solution, pH 7.4, 2 h at R.T., dehydrated in a graded ethyl alcohol series followed by propylene oxide, and embedded in Agar 100 resin. Polymerization was carried out at 60 °C for 48 h. Ultrathin sections were cut on an LKB III ultratome, mounted on copper 200 mesh thin bar grids and stained with uranyl acetate and lead citrate. Pictures were captured on a Tecnai-12 electron microscope (Phillips, The Netherlands) equipped with Megaview II CCD camera and 3.0 analysis software (Soft Imaging System, GmbH, Germany).

A 1 cm square of dorsal skin was excised from the mouse, placed flat on a piece of paper towel and fixed in 4% paraformaldehyde and 2% glutaraldehyde in 0.05M cacodylate buffer (pH 7.4) overnight at room temperature. The tissues were then rinsed four times for 10 min each in cacodylate buffer and post-fixed and stained with 1% osmium tetroxide and 1.5% potassium ferricyanide in 0.1 M cacodylate buffer for 1 h. Tissues were then washed four times in cacodylate buffer followed by dehydration once for 10 min each in ethanol at 30%, 50%, 70%, 80%, 90% and 95% followed by three times for 20 min each in 100% anhydrous ethanol, and twice for 10 min each in propylene oxide. Following dehydration, the tissues were infiltrated with increasing concentrations of Agar 100 resin (20%, 50%, 75% and 100%) in propylene oxide for 16 h per step. The tissues were then embedded in fresh resin and allowed to polymerize in a 60°C oven for 48 h. The embedded tissue blocks were sectioned with a diamond knife on a Leica Reichert Ultracut S microtome, and ultrathin sections (80 nm) were collected onto 200 mesh, carbon/formvar-coated copper grids. The sections were then sequentially stained with uranyl acetate and lead citrate for 10 min each and viewed with a Tecnai 12 100 kV TEM (Phillips) equipped with a MegaView II CCD camera and Analysis® version 3.0 software (SoftImaging System).