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Mammalian cells change volume during mitosis.

Boucrot E, Kirchhausen T - PLoS ONE (2008)

Bottom Line: Using single cell-imaging methods we have found that the volume of adherent cells grown in culture decreases as the cells rounds when it enters mitosis.Rapid volume recovery initiates before abscission as cells make the transition from metaphase to cytokinesis.These volume changes are simultaneous with the rapid surface area decrease and recovery observed in mitotic cells [1].

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Immune Disease Institute, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT
Using single cell-imaging methods we have found that the volume of adherent cells grown in culture decreases as the cells rounds when it enters mitosis. A minimal volume is reached at metaphase. Rapid volume recovery initiates before abscission as cells make the transition from metaphase to cytokinesis. These volume changes are simultaneous with the rapid surface area decrease and recovery observed in mitotic cells [1].

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Single cell imaging during mitosis.(A) Representative images of the same BSC1 cell visualized by bright field phase contrast illumination during prophase, metaphase or cytokinesis (B) Ortogonal fluorescent views corresponding to the distribution of EGFP-CAAX in the same cell imaged using the confocal spinning head. The stacks along the z-axis correspond to sequential optical sections acquired 0.25 µm apart. The panels show the fluorescence signal along the z-axis (top) along a single confocal plane positioned approximately in the middle of the cell. Bar, 10 µm. (C) The mask (light blue) corresponds to the position of the EGFP-CAAX signal calculated for each confocal plane (see Methods). The outline of the outer boundary determined for any given plane defines the enclosed area for such plane (dark blue). (D) Three-dimensional rendition comparing the distribution of EGFP-CAAX (green) and the calculated cell volume (blue).
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pone-0001477-g001: Single cell imaging during mitosis.(A) Representative images of the same BSC1 cell visualized by bright field phase contrast illumination during prophase, metaphase or cytokinesis (B) Ortogonal fluorescent views corresponding to the distribution of EGFP-CAAX in the same cell imaged using the confocal spinning head. The stacks along the z-axis correspond to sequential optical sections acquired 0.25 µm apart. The panels show the fluorescence signal along the z-axis (top) along a single confocal plane positioned approximately in the middle of the cell. Bar, 10 µm. (C) The mask (light blue) corresponds to the position of the EGFP-CAAX signal calculated for each confocal plane (see Methods). The outline of the outer boundary determined for any given plane defines the enclosed area for such plane (dark blue). (D) Three-dimensional rendition comparing the distribution of EGFP-CAAX (green) and the calculated cell volume (blue).

Mentions: Cells entering prophase were readily identified by the appearance of their chromosomes observed using bright field phase contrast illumination. The spatial distribution of EGFP-CAAX of a selected cell was immediately determined by a first round of three-dimensional fluorescence imaging, followed by a second imaging series after the cell reached metaphase, and a final one during cytokinesis, ∼45 min after the onset of anaphase, at time at which telophase has ended but abscission has not yet occurred. Representative examples of images from a cell (out of 10 analyzed) at these three stages are shown in Figure 1 (bright field (A) and fluorescence (B)). We used the outline of the EGFP-CAAX signal (Figure 1B, green) defined by the most external pixels to generate a mask (Figure 1C, blue). Three-dimensional rendering of the data shows that the masking procedure faithfully follows the cell boundary (Figure 1D, green) used to define the enclosed volume (Figure 1D, blue).


Mammalian cells change volume during mitosis.

Boucrot E, Kirchhausen T - PLoS ONE (2008)

Single cell imaging during mitosis.(A) Representative images of the same BSC1 cell visualized by bright field phase contrast illumination during prophase, metaphase or cytokinesis (B) Ortogonal fluorescent views corresponding to the distribution of EGFP-CAAX in the same cell imaged using the confocal spinning head. The stacks along the z-axis correspond to sequential optical sections acquired 0.25 µm apart. The panels show the fluorescence signal along the z-axis (top) along a single confocal plane positioned approximately in the middle of the cell. Bar, 10 µm. (C) The mask (light blue) corresponds to the position of the EGFP-CAAX signal calculated for each confocal plane (see Methods). The outline of the outer boundary determined for any given plane defines the enclosed area for such plane (dark blue). (D) Three-dimensional rendition comparing the distribution of EGFP-CAAX (green) and the calculated cell volume (blue).
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Related In: Results  -  Collection

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pone-0001477-g001: Single cell imaging during mitosis.(A) Representative images of the same BSC1 cell visualized by bright field phase contrast illumination during prophase, metaphase or cytokinesis (B) Ortogonal fluorescent views corresponding to the distribution of EGFP-CAAX in the same cell imaged using the confocal spinning head. The stacks along the z-axis correspond to sequential optical sections acquired 0.25 µm apart. The panels show the fluorescence signal along the z-axis (top) along a single confocal plane positioned approximately in the middle of the cell. Bar, 10 µm. (C) The mask (light blue) corresponds to the position of the EGFP-CAAX signal calculated for each confocal plane (see Methods). The outline of the outer boundary determined for any given plane defines the enclosed area for such plane (dark blue). (D) Three-dimensional rendition comparing the distribution of EGFP-CAAX (green) and the calculated cell volume (blue).
Mentions: Cells entering prophase were readily identified by the appearance of their chromosomes observed using bright field phase contrast illumination. The spatial distribution of EGFP-CAAX of a selected cell was immediately determined by a first round of three-dimensional fluorescence imaging, followed by a second imaging series after the cell reached metaphase, and a final one during cytokinesis, ∼45 min after the onset of anaphase, at time at which telophase has ended but abscission has not yet occurred. Representative examples of images from a cell (out of 10 analyzed) at these three stages are shown in Figure 1 (bright field (A) and fluorescence (B)). We used the outline of the EGFP-CAAX signal (Figure 1B, green) defined by the most external pixels to generate a mask (Figure 1C, blue). Three-dimensional rendering of the data shows that the masking procedure faithfully follows the cell boundary (Figure 1D, green) used to define the enclosed volume (Figure 1D, blue).

Bottom Line: Using single cell-imaging methods we have found that the volume of adherent cells grown in culture decreases as the cells rounds when it enters mitosis.Rapid volume recovery initiates before abscission as cells make the transition from metaphase to cytokinesis.These volume changes are simultaneous with the rapid surface area decrease and recovery observed in mitotic cells [1].

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Immune Disease Institute, Harvard Medical School, Boston, Massachusetts, USA.

ABSTRACT
Using single cell-imaging methods we have found that the volume of adherent cells grown in culture decreases as the cells rounds when it enters mitosis. A minimal volume is reached at metaphase. Rapid volume recovery initiates before abscission as cells make the transition from metaphase to cytokinesis. These volume changes are simultaneous with the rapid surface area decrease and recovery observed in mitotic cells [1].

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