Correlative time-lapse imaging and electron microscopy to study abscission in HeLa cells.
HeLa cells are widely used as a model system to study cell division. The last step of cell division, abscission, occurs at an about 1 μm wide intercellular bridge that connects the post-mitotic sister cells. Abscission often occurs long after ingression of the cleavage furrow, and no efficient methods to synchronize cells to this stage are available. Here, we have developed a correlative fluorescence time-lapse imaging and electron microscopic approach using Aclar sheets with engraved grid patterns. This grid pattern, leaving a negative imprint on thin-layer embedded samples, allows identification of cells selected from the time-lapse imaging for serial-section electron microscopy. This method facilitates the ultrastructural analysis of specific stages of abscission.
Guizetti
Julien
J
Institute of Biochemistry, Swiss Federal Institute of Technology Zurich (ETHZ), CH-8093 Zurich, Switzerland.
Mäntler
Jana
J
Müller-Reichert
Thomas
T
Gerlich
Daniel W
DW
D002462:Cell Membrane / Q000648:ultrastructure; D048749:Cytokinesis; D003599:Cytoskeleton / Q000648:ultrastructure; D006367:HeLa Cells / Q000502:physiology / Q000648:ultrastructure*; D006651:Histocytochemistry / Q000379:methods; D006801:Humans; D008854:Microscopy, Electron / Q000295:instrumentation / Q000379:methods*; D059008:Time-Lapse Imaging / Q000295:instrumentation / Q000379:methods*