Ds ri2 camera

Lung tissue sections were processed for immunohistochemical (IHC) staining as described previously [39 (link)]. Briefly, after the deparaffinization and antigen repossession step, the endogenous peroxidase activity was blocked by dipping the slides in a 1:100 dilution of 30% hydrogen peroxide (Sigma) in a methyl alcohol solution for 15 min. After washing, the unbinding sites were blocked with BSA followed by adding LC3-II and Parkin purified rabbit anti-mouse antibodies at a concentration of 1:50 and then incubated overnight at 4 °C. After incubation, the sections were washed with PBS thrice for 5 min each, followed by incubation for 90 min at 37 °C with HRP-labeled secondary antibodies. After washing, enzymatic activity was revealed by using 3, 3′-Diaminobenzidine (DAB). Digital images were collected on an Olympus DP72 microscope fitted with DS-Ri2 camera. To quantify the intensity of LC3 and Parkin, 3 lung sections from independent animals of each group were visualized under low (40×) and high (100×) power of magnifications. The stained area compared with the total tissue area was determined by using Image-J software.

Gross and histological studies were carried out to evaluate the effect of nilotinib treatment on different organs of M. bovis infected mice. Lungs, spleens, kidneys, and livers from the sacrificed animals were collected under a sterile environment as soon as possible [32 (link)]. For gross pathology evaluation, lungs and spleens were weighed and clear images were captured at different time points of infection. For histological study, tissues were fixed in a 10% formaldehyde solution, embedded in paraffin, and cut into sections using a microtome (Leica RM2235; Leica, Buffalo Grove, IL, USA). Tissue was mounted on glass slides, deparaffinized, and stained with: haematoxylin and eosin (H&E) or Ziehl-Neelsen (ZN) for visualizing the acid-fast M. bovis bacilli. Sections stained with H&E or ZN were observed under low (10× or 20×) and high magnification (40× or 100×) by using an Olympus DP72 microscope fitted with a DS-Ri2 camera (Olympus, Instruments Inc., Tokyo, Japan).