Bx63f upright microscope

For gut morphological studies, adult C. longicaudata and T. domestica were anesthetized for 10 min at 4°C and dissected under a Zeiss Stemi 2000-C stereo microscope (Carl Zeiss Microscopy, LLC, Thornwood, NY). The gut was carefully dissected from the rest of the body and images were taken with a Canon DS126311 camera (Canon, Ota, Tokyo, Japan) mounted on the stereo microscope. Adult C. longicaudata and T. domestica for histological studies were sacrificed by incubation at -20°C for ten minutes and then fixed in Carnoy’s solution (60% ethanol, 30% chloroform, and 10% glacial acetic acid) for four hours at 4°C. After fixing, whole insects were transferred to 70% ethyl alcohol and sent to the Biomedical and Diagnostic Services, University of Tennessee College of Veterinary Medicine (Knoxville, TN) for sectioning and staining with hematoxylin and eosin. Histological sections were examined and documented using an Olympus BX63F upright microscope (Olympus Corporation, Shinjuku, Tokyo, Japan).

The cell lines SW480 and RKO were seeded in 12-well plates with one coverslip per well. 24 h after seeding, the treatments were processed as described in Section 2.6. 48 h later the cells were fixed and stained as previously reported by us [36 (link)]. Representative images were obtained using a fluorescence microscope (Olympus motorized BX63F Upright Microscope) at a magnification of 600×.

SW480 cell line was seeded at a concentration of 1 × 10⁵ cells/ml, in 12-well plates with one coverslip per well. After 24 h, the cells were exposed to the IC₅₀ values of iron compounds and cisplatin, and the cells of negative control were treated with DMSO 0.1%. After 48 hours of incubation, the cells were washed twice with PBS and fixed with paraformaldehyde 4% (w/v) for 10 minutes. After fixation, cells were incubated with NH₄Cl 50 mM and washed twice with PBS, for five minutes. The cells were permeabilized using Triton X-100 0.2%, for five minutes, and blocked with PBS–BSA 3% for 20 minutes. Subsequently, cells were incubated with Alexa Fluor™ 568 Phalloidin (ThermoFisher Scientific®, Waltham, MA, USA), diluted in PBS (1:40), for one hour in the dark, and washed twice with PBS. To finalize, coverslips were mounted using 5 μL of VECTASHIELD Antifade Mounting Medium with DAPI (Vector Laboratories^®, Peterborough, UK) in microscope slides. The results were obtained from at least three independent experiments. Representative images were obtained in a fluorescence microscope (Olympus motorized BX63F Upright Microscope) (Olympus©, Tokyo, Japan) at a magnification of 600×.