Axiocam 208 color digital camera

At the end of each day of experiment, the teeth were taken out from the artificial saliva (control group) and from the I-131 solution (experimental groups), washed with water, dried and then stored in 10% phosphate-buffered formalin, for 48 h. After this process, within each group, 3 specimens were taken out; the roots were embedded in self-cured acrylic resin templates, up to 2 mm below cementoenamel junction, in order to simulate the alveolar bone. When the resin was ready, each specimen was taken out from the template and each crown was sectioned longitudinally in the mesio-distal direction resulting in two parts (buccal, lingual) using a diamond saw cooled with water mounted in a precision saw (Isomet, Buehler, Lake Bluff, IL, USA) resulting in sections of 1 mm width. For histological examination, an Axio Scope A1 microscope (Zeiss, Oberkochen, Germany) was used. The photomicrographs were taken using an Axiocam 208 color digital camera and ZEN core imaging software from Zeiss.

Brains were fixed in 4% paraformaldehyde for 3 days and cut coronally into five divisions of similar thickness with a scalpel. Each brain division was embedded in paraplast and cut coronally to sections of 1 µm thickness using a microtome (HM 400, Microm, Berlin, Germany). First, one section per division was stained with hematoxylin-eosin, and the number of lesions was counted in the entire section. If a lesion was found, the consecutive section was examined for the presence of microglia by immunohistochemistry using an antibody for Iba1 (2-MI004-10, Quartett, Berlin, Germany). Hematoxylin-eosin staining and immunohistochemistry were performed as described previously in [36 (link)–38 (link)]. An Olympus BX46 light microscope (Olympus, Shinjuku, Japan) equipped with an Axiocam 208 color digital camera (Zeiss, Oberkochen, Germany) and ZEN 2.6 (Zeiss, Oberkochen, Germany) imaging software was used to examine histological sections.