Critical point dryer e3000 series

Mouse tracheae were taken from the AAV5‐CON305 and AAV5‐CTNNAL1‐RNAi mice groups. Subsequently, the tissues were fixed in cold (4℃) 3% glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 7.4). After 1 h fixation, the samples were trimmed and then transferred to the fresh fixative at 4℃ for another 3 h. The samples were washed in cold (4℃) dilute buffer (equal volume of 0.1 M sodium cacodylate buffer and distilled water); dehydrated in cold condition (4℃) with graded ethyl alcohol in ascending concentrations, ethyl alcohol and acetone in the ratios 3:1, 1:1 and 1:3 and with anhydrous acetone and dried using Critical Point Dryer (E3000 series; Quorum Technologies, East Sussex, UK) with liquid carbon dioxide as the transitional fluid. The tissues were then attached to stubs, coated with gold using Sputter Coater (SC7620; Quorum Technologies) and were examined with a Scanning Electron Microscope (EVO LS 10; Carl Zeiss, Oberkochen, Germany).

Tissue samples excised from fish of the groups I, II and III, were rinsed in physiological saline, dipped briefly in a 0.1% solution of S-carboxymethyl-L-cysteine (sigma Aldrich) to remove mucus following Whitear and Moate (1994) [20 ] and then fixed in cold (4 °C) 3% glutaraldehyde (Lifeline Medical, Inc) in 0.1 M sodium cacodylate buffer (pH 7.4) (sigma Aldrich) for 4 h. After fixation, the tissue samples were dehydrated, dried using Critical Point Dryer (E3000 series; Quorum Technologies), attached to stubs and then coated with gold using Sputter Coater (SC7620; Quorum Technologies) following Verma et.al.(2017) [21 (link)]. Processed samples were examined under a JEOL 5800LV scanning electron microscope.