Coolscope digital light microscope

The tissue samples collected on 10% neutral buffered formalin were fixed for 48 h at 4 °C. The fixed samples were passed into a serial ascending grade of ethanol and xylene and embedded in paraffin wax. Then, 5-μm sections were cut and stained with hematoxylin and eosin (H&E) [26 ]. In brief, the sections were deparaffinized in three changes of xylene, rehydrated through a descending series of ethanol, and stained with hematoxylin. The slides were washed in tap water and then stained with eosin, followed by washing in tap water and rinsing in distilled water. The sections were dehydrated in ethanol, cleared in xylene, and mounted. The stained sections were examined using Coolscope Digital light Microscope (Nikon, Japan).

Samples of the lung were fixed in 4% formaldehyde. Sectioning was done after the samples were embedded in paraffin wax. Grocott's methenamine silver (GMS) was utilized to stain the lung samples for the detection of fungus. For the histological analysis procedure, lung samples were stained with either hematoxylin and eosin (H&E). Analyzed through the use of COOLSCOPE digital light microscope (Nikon Co., Tokyo, Japan), lung injury was scored according to criteria defined by Mikawa et al. [22 (link)] as follows: (1) alveolar hyperemia, (2) hemorrhage, (3) interstitial or aggregation of interstitial or neutrophils, and (4) thickening of the alveolar septum or hyaline membrane formation. Pneumoniae pulmonary infection scores were approximated through the method by the scoring standard published by Cimolai et al. [23 (link)]. The scoring standard is based on (1) the infiltration degree of inflammatory cells around the trachea and bronchiole 0–3, (2) quality of trachea and bronchiole infiltrate 0–3, (3) infiltration degree of inflammation in trachea and bronchiole cavity 0–2, (4) infiltration of inflammatory cells around blood vessels, degree 0–3, and (5) inflammation of the lung parenchyma which involves the range 0, 3, and 5. The severity of the inflammation is directly proportional to the magnitude of the score.