Ds fi1 u2 camera

The OxiSelect™ Comet Assay Kit (Cell Biolabs, San Diego, USA) was used. TM3 cells, at a density of 1 × 10⁵/mL, were washed and suspended in 10% OxiSelect™ Comet Agarose. Then, 75 μL of the suspension was pipetted into each well of the OxiSelect™ 3-Well Comet Slides. According to the manufacturer’s instructions, the slides were processed and finally electrophoresed at 1.3 V/cm for 20 min. The comets stained with ethidium bromide (20 µg/mL) were imaged using a Nikon (Tokio, Japan) Eclipse Ci-L epifluorescence microscope, and at least 70 cells per slide for each group were recorded using the Nikon DSFi1-U2 camera equipped with the Nikon NISF software. The etoposide-treated cells (350 µM, Sigma-Aldrich, Burlington, MA, USA) were used as a positive control. The DNA damage was analysed using the CaspLab^® software (2020, http://casplab.com, (accessed on 15 August 2021)). The percentage of DNA in the comet tail was quantified, and the tail moment was calculated as the percentage of DNA in the tail multiplied by the length of the tail. Additionally, the nuclei of the TM3 cells were classified into five categories based on the quantity of DNA in the tail [86 (link)].

(i) H&E and PAS-AB. Mouse colons were placed intact in cassettes and fixed in 10% Carnoy’s fixative. Paraffin-embedded tissue sections (7 μm) were processed for hematoxylin and eosin (H&E) or periodic acid-Schiff/Alcian blue (PAS-AB) staining. H&E and PAS-AB sections were examined by bright-field and imaged on the Nikon Eclipse 90i (Nikon) microscope using a DS-Fi1-U2 camera (Nikon) with a differential interference contrast (DIC) objective.
(ii) Immunofluorescence.F. nucleatum localization was examined using a Fusobacterium-specific FISH probe (5′-CGCAATACAGAGTTGAGCCCTGC-3′), and total bacteria were examined using a universal bacterial FISH probe EUB338 (5′-GCTGCCTCCCGTAGGAGT-3′; Integrated DNA Technologies [IDT]) (110 (link)). Briefly, tissue sections were dehydrated and incubated with the Fusobacterium probe at 45°C in a dark humidifying chamber, hybridized for 45 min, and counterstained with MUC2 (1:200 dilution; Cloud-Clone Corp., PAA705Mu01) and Hoechst 33342 (Invitrogen, H3570). Immunostained slides were imaged on an Eclipse 90i (Nikon, Tokyo, Japan) with a 20× Plan Apo (NA 0.75) DIC objective, and the images were recorded using a CoolSNAP HQ2 camera (Photometrics) using a Nikon Intensilight C-HGFI mercury lamp.