The samples were fixed in 7% formaldehyde for 5 days, after which the samples were oriented and placed in cassettes. Tissue processing was performed using a vacuum infiltration processor, Tissue-Tek VIP 5 Jr (Sakura, Alphen aan den Rijn, Netherlands). Paraffin embedding and sectioning were performed using the Tissue-Tek TEC 6 system (Sakura, Alphen aan den Rijn, Netherlands) and Accu-Cut SRM 200 Rotary Microtome (Sakura, Alphen aan den Rijn, Netherlands). Slide staining was performed using the automated slide stainer Tissue-Tek Prisma Plus (Sakura, Alphen aan den Rijn, Netherlands), according to the internal staining protocol, using Mayer Modified Hematoxylin (Titolchimica, Rovigo, Italy) and Eosine solution (10 g Eosine B in 1000 mL distilled water).
Immunohistochemistry was performed automatically on 3 μm thick sections of formalin-fixed and paraffin-embedded tissues with MD Stainer (Vitro Master Diagnostica®, Granada, Spain) using ethylenediaminetetraacetic acid (EDTA), at pH = 9, for antigen retrieval. We used anti-SARS coronavirus NP mouse anti-virus antibody (clone B46F, Invitrogen, Waltham, MA, USA) at a 1:100 dilution for the immunohistochemical assessment. Positive cells are colored in brown.
Microscopic examination was performed by the same experienced pathologist (D.G.), using a Leica DM1000 clinical microscope (Leica, Wetzlar, Germany) with a dedicated image acquisition camera and software. All sections were examined by the same experienced investigator (D.G.).
Immunohistochemistry was performed automatically on 3 μm thick sections of formalin-fixed and paraffin-embedded tissues with MD Stainer (Vitro Master Diagnostica®, Granada, Spain) using ethylenediaminetetraacetic acid (EDTA), at pH = 9, for antigen retrieval. We used anti-SARS coronavirus NP mouse anti-virus antibody (clone B46F, Invitrogen, Waltham, MA, USA) at a 1:100 dilution for the immunohistochemical assessment. Positive cells are colored in brown.
Microscopic examination was performed by the same experienced pathologist (D.G.), using a Leica DM1000 clinical microscope (Leica, Wetzlar, Germany) with a dedicated image acquisition camera and software. All sections were examined by the same experienced investigator (D.G.).
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