Microscope axioscope 5 7 kmat

Liver tissue sections (5 μm) were obtained from cryostat and stained with hematoxylin and eosin. Following staining, the slides were observed with an optical microscope (Microscope Axioscope 5/7 KMAT, Carl Zeiss, Oberkochen, Germany) connected to a digital camera (Microscopy Camera Axiocam 208 color, Carl Zeiss). The morphological evaluation of the liver steatosis was performed by a medical doctor specialized in anatomic pathology (FR), using a semiquantitative scoring system identified for NAFLD (22 (link)). This score was also used for inflammation and fibrosis that we did not observe. According to this score, steatosis was graduated from 0 to 3 where 0: 0–5%, 1: 3%−33%, 2: 34%−66%, and 3: >66%. All the observations were performed in high-power fields (HPF; magnification 400 ×), and the arithmetic means of percentage was used for statistical analyses.

Hepatic samples were immediately stored in paraformaldehyde for 48 h, then were moved to a 20% PBS/sucrose solution, and, after 1 week, to 10% PBS/sucrose solution. At last, the solution was removed and samples were stored at −80°. Liver tissue sections (5 µm) were obtained from cryostat and stained with hematoxylin and eosin. Following staining, the slides were observed with an optical microscope (Microscope Axioscope 5/7 KMAT, Carl Zeiss, Oberkochen, Germany) connected to a digital camera (Microscopy Camera Axiocam 208 colour, Carl Zeiss). For the steatosis evaluation, a semiquantitative analysis was performed by two independent observers in a high-power field (HPF) (magnification 400×) and repeated for 10 HPFs.

For histological analysis, lung samples from six COVID-19 subjects (COVID-19 group) and six control subjects (control group) were selected. All samples were fixed in formalin and embedded in paraffin. Sections of 4–5 µm thickness were prepared from all paraffin blocks using a cutting microtome. These sections, placed on slides, were dewaxed in xylene for 10 min and rehydrated by sequential immersion in a descending scale of alcohols and transitioned in water for five minutes. The slides were then stained with hematoxylin and eosin, mounted with coverslips, and finally observed with an optical microscope (Microscope Axioscope 5/7 KMAT, Carl Zeiss, Milan, Italy) connected to a digital camera (Microscopy Camera Axiocam 208 color, Carl Zeiss, Milan, Italy).

Immunohistochemical (IHC) investigations were carried out on 5 µm thick sections obtained from paraffin blocks using a cutting microtome. The IHC reactions were performed using the automated IHC system of the Biotechnology Laboratory of the Euro-Mediterranean Institute of Sciences and Technologies (IEMEST) (IntelliPath Flx, Biocare Medical, distributed by Bio-Optica, Milan, Italy). The primary antibodies used are indicated in Table 2. At the end of the immunostaining cycle, the slides were prepared for observation with coverslips using a permanent mounting medium (VectaMount, Vector, H-5000, Burlingame, CA, USA). Observation of the sections was performed with an optical microscope (Microscope Axioscope 5/7 KMAT, Carl Zeiss, Milan, Italy) connected to a digital camera (Microscopy Camera Axiocam 208 color, Carl Zeiss, Milan, Italy). Two independent pathologists (F.C. and F.R.) examined the specimens on two separate occasions and performed a semiquantitative analysis to determine the percentage of cells positive for Hsp60 and Hsp90. The percentage of immunopositivity was evaluated in a high-power field (HPF) at 400× magnification and repeated for 10 HPF. The average of the percentages of all immunosemiquantifications performed in each case for the two groups was considered as a conclusive result, and this value was used for the statistical analysis.