Biozero bz 9000e

Skin samples were fixed in 4% buffered formalin and embedded in paraffin. Four µm thick sections of the ears were cut and stained for H&E as previously described^{70 (link)}. Cryostat sections of the ear skin were stained for granulocytes. Briefly, a monoclonal antibody against Gr-1 (Ly6C/G, RBC-8C5, eBioscience) was used as the primary antibody, and an alkaline phosphatase-conjugated goat anti-rat IgG (Roth, Karlsruhe, Germany) comprised the secondary antibody. A Biozero BZ-9000E from Keyence (Keyence, Osaka, Japan) was used for light microscopy.

Following surgical resection, lung tissue specimens were immediately fixed in 10% formalin (BioVitrum, Moscow, Russia). Dehydration was carried out according to the standard protocol in a series of alcohols from 70 to 96%, 100% ethanol was replaced by isopropyl alcohol. After dehydration with isopropyl alcohol, the samples were embedded in paraffin Hystomix (BioVitrum, Moscow, Russia), and then were cut at 5 μm thickness for preparation of serial section slides. Tissue sections were stained with hematoxylin & eosin (H&E) and embedded in Canada balsam for histological examination. Slides were used to classify the clinical material into tuberculoma with high and moderate activity of TB inflammation. A Biozero BZ-9000E from Keyence (Keyence, Osaka, Japan) was used for light microscopy. The research was carried out using the equipment of the Core Centrum of the Institute of Developmental Biology RAS.