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Fig. 2 | Biology Direct

Fig. 2

From: A link between mitotic defects and mitotic catastrophe: detection and cell fate

Fig. 2

Representative images of morphological changes after induction of mitotic catastrophe were received using classical fluorescent confocal microscopy or imaging flow cytometry. A The ovarian carcinoma Caov-4 cells and the lung adenocarcinoma U1810 cells were treated with colcemid 10 ng/mL, a chemotherapeutic agent that depolymerizes microtubules, for 48 h. Cells were grown, treated, fixed in 4% paraformaldehyde, and stained with MitoTracker Red FM and DAPI directly on coverslips. Mitotic catastrophe development was evaluated by analysis of nuclear morphology using a 63 × /1.4 oil objective lens of a LSM 780 confocal laser scanning microscope (Zeiss). After the colcemid treatment of the cells the multi- and micronucleation were detected. B The ovarian carcinoma Caov-4 cells were treated with colcemid 10 ng/mL for 48 h. After incubation, the cells were collected by trypsin–EDTA and transferred to conditioned medium. The cells were centrifuged and washed twice with cold phosphate-buffered saline. Then, the cells (2.0 × 105) were resuspended in PBS and DAPI was added to the sample and incubated in the dark at 4 °C for 15 min. After incubation, the cells were analyzed by imaging flow cytometry with the ImageStream Mark II - AMNIS (Millipore Merck, Germany). After the colcemid treatment of Caov-4 cells the multinucleation was detected

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