The slides were fixed and stained with ProLong[R] Gold Antifade Mountant
and DAPI (Thermo Fisher Scientific, USA), which stained the nuclei in blue, and were analyzed under a confocal microscope (DMI 6000; Leica, German).
Mount it with DPX mountant
and visualize under microscope.
The sections were then stained with standard Haematoxylin and Eosin method and then mounted (DPX mountant
They were then serially sectioned to 5 [micro]m thickness using microtomy, stained with Hematoxylin and Eosin (H&E) stain and were fixed on microscopic slides using DPX mountant
. The slides were then examined under a Motic DMB1 2MP light microscope (China) at 40X magnification for treatment-induced structural alterations and the photomicrographs were snapped.
The fungi were mounted on the clean glass slide with lactophenol, sterile distilled water or lactophenol cotton blue stain and edges were sealed with DPX mountant
The slides were visualized with diaminobenzine (DAB), counterstained by hematoxylin, dehydrated, and mounted in a neutral resin mountant
. Blue staining, as determination of TRIF protein expression, was displayed and examined using Nikon E800 microscope according to the manufacturer's instructions.
The cover slips were dried and placed on a labeled clean glass slides using a mountant
(DPX) for fluorescence microscopic examination (Leica Microsystems Limited, Heerbrugg, Switzerland) of VP2 antigen positive cells .
The chemicals used were xylene, DPX mountant
, euthatal, methanol (MetOH), distilled water, 4% paraformaldehyde, phosphate-buffered saline (PBS), 0.2% Triton X-100 in Trizma base solution (TXTBS), 3% normal horse serum (NHS; S-2000; Vector Laboratories) in TXTBS, 5% normal goat serum (NGS; S-1000; Vector Laboratories) in TXTBS, and Trizma base (TB) and Trizma nonsaline (TN) solutions.
The segments were mounted with ProLong Gold antifade Mountant
reagent (Invitrogen, Carlsbad, CA, USA) on a glass slide.
Monolayers were mounted in ProLong Gold Antifade Mountant
(Thermo Fisher Scientific-Molecular Probes) and imaged by confocal microscopy (Leica TCS SP5 Confocal Microscope: Leica Microsystems, Mannheim, Germany).
After dehydration and clearing, sections were coverslipped with DPX mountant