## 3 Results
### Nuclear Morphology of Non-Irradiated (Control) and Irradiated H1299 and MCF-10A Cells  
Effects of W-band MMW irradiation on nuclei number, shape, and area were examined using DAPI (Figure 2A) and analyzed by observations under a fluorescent microscope. Concomitant with our recent report [19] observations in this study showed that most of the non-irradiated H1299 lung cancer cells present an average cellular size of 1500 \(\upmu\)m\({}^{2}\) containing a single large round or oval nuclei  
Figure 1: Experimental Schematic of H1299 human lung cancer cells monolayer irradiated by millimeter wave (MMW) for 2, 4, and 10 min in 35 mm dishes. The effects observed in the irradiated cells were compared to two types of control cells (see methods)—(i) the cells that remained in the incubator during the entire experiment, and (ii) the cells that were transferred to the irradiation setup and brought back together with the experimental groups but were not irradiated. (Figure 2A). One to four percent of the cells were 2.5-3.5 times larger (5000 \(\upmu\)m\({}^{2}\)) and characterized with multi-lobed nuclei. These polyploid cells contained 3-11 nuclei per cell. Cells treated with MMW showed changes in nuclear morphology with the 4 min regime having a greater impact than the 2 min regime. The nuclei of cancer cells play a key role in the assessment of tumor malignancy through changes in genetic structure [20]. Cancer cells are characterized by a large nucleus with an irregular size and shape as well as prominent nucleoli. Morphological changes in their areas (in the adherent state) and the shapes of the cells lead to changes in the cell's internal structures (like the nuclei) and increase the chromosomal instability (ploidy) of cancer cells [21]. Non-tumorigenic MCF-10A cells were also irradiated under the same parameters (Figure 2B). Interestingly, even at 16 min of MMW treatment nuclear morphology of MCF-10A cells remained unaltered relative to that of control, in comparison to the effects observed for 4 min of MMW exposure on H1299 cells. The nuclear sizes of the H1299 cells in normal growth conditions were measured, over 14 days of the experimental observation period (Figure 2C), after processing them through the same conditions as the irradiated cells except without the MMW treatment procedure itself. The average areas of control H1299 cell nuclei ranged from 100-250 \(\upmu\)m\({}^{2}\) (subgroups 1-5), 9% of the cells contained nuclei under division (subgroup 6-7) with an area 200-300 \(\upmu\)m\({}^{2}\) and up to 4% of the cells (subgroups 8-10) contained a multi-lobular, irregular nuclei, or cells that contained 3-11 nuclei per cell, with area ranging 300-600 \(\upmu\)m\({}^{2}\). No significant changes in area distribution were observed over seven days of proliferation, while the population of the H1299 cells with larger nuclei (subgroups  
Figure 2: The distribution of the nuclei sizes and shapes in control and treated cells in response to MMW irradiation. (**A**) Representative images of H1299 nuclei morphology before and after irradiation. (a) Control cells’ nuclei show circle or oval morphology. Numbers indicate the subgroup classification as described in Table 1; (b) kidney-like nuclei appeared over long-term incubation (7–14 days) after irradiation for 2 min; and (c) multi-lobed and poly-nuclear cells appeared over long-term incubation (7–14 days) after irradiation for 4 min. Scale bars correspond to 10 \(\upmu\)m. (**B**) Representative images of MCF-10A nuclei morphology before and after irradiation. (a) Control cells’ nuclei show circle or oval morphology; (b–d) nuclei shape remains same over long-term incubation (7–14 days) after irradiation for 4, 8, and 16 min. Scale bars correspond to 10 \(\upmu\)m. (**C**) Control H1299 cell population distribution by area size of nuclei in the cells. The areas indicated are given in \(\upmu\)m\({}^{2}\). Analysis of nuclei distribution in the control groups consisted of four dishes (Ra–d).