A simple and effective method for ultrastructural analysis of mitosis inDrosophilaS2 cells.
TheDrosophilaS2 tissue culture cells are a widely used system for studies on mitosis. S2 cells are particularly sensitive to gene silencing by RNA interference (RNAi), allowing targeted inactivation of mitotic genes. S2 cells are also well suited for high-resolution light microscopy analysis of mitosis in fixed cells, and can be easily immunostained to detect mitotic components. In addition, S2 cells are amenable to transformation with plasmid encoding fluorescently tagged mitotic proteins, allowingin vivoanalysis of their behavior throughout cell division. However, S2 cells have not been widely used for transmission electron microscopy (TEM) analysis, which provides ultrastructural details on the morphology of the mitotic apparatus that cannot be obtained with high-resolution confocal microscopy. Here, we describe a simple method for the ultrastructural analysis of mitosis inDrosophilaS2 cells. •Our method, which involves fixation and sectioning of a cell pellet, provides excellent preservation of mitotic structures and allows analysis of a higher number of mitotic divisions per sample, compared to correlative light-electron microscopy.•Dividing cells are randomly oriented within the pellet and are sectioned along different planes, providing all-around information on the structure of the mitotic apparatus.
Institute of Molecular and Cellular Biology, Siberian Branch of RAS, Novosibirsk, 630090, Russia; Institute of Cytology and Genetics, Siberian Branch of RAS, Novosibirsk, 630090, Russia.
Anton
A
Strunov
Institute of Molecular and Cellular Biology, Siberian Branch of RAS, Novosibirsk, 630090, Russia.
Lidiya V
LV
Boldyreva
Institute of Molecular and Cellular Biology, Siberian Branch of RAS, Novosibirsk, 630090, Russia; Kazan Federal University, Kazan, 420008, Russia.
Gera A
GA
Pavlova
Institute of Molecular and Cellular Biology, Siberian Branch of RAS, Novosibirsk, 630090, Russia; Institute of Cytology and Genetics, Siberian Branch of RAS, Novosibirsk, 630090, Russia; Novosibirsk State University, Novosibirsk, 630090, Russia.
Alexey V
AV
Pindyurin
Institute of Molecular and Cellular Biology, Siberian Branch of RAS, Novosibirsk, 630090, Russia; IBPM CNR and Department of Biology and Biotechnology, Sapienza University of Rome, Rome, 00185, Italy.
Maurizio
M
Gatti
Institute of Molecular and Cellular Biology, Siberian Branch of RAS, Novosibirsk, 630090, Russia; Institute of Cytology and Genetics, Siberian Branch of RAS, Novosibirsk, 630090, Russia.
Elena
E
Kiseleva
A simple and effective method for ultrastructural analysis of mitosis in Drosophila S2 cells; Drosophila S2 cells; Mitosis; Transmission electron microscopy; Ultrastructural analysis; Unsynchronized cell cultures