RRC ID 85540
Author Murata T, Tsutsumi M, Otomo K, Nemoto T.
Title Cell Division Imaging of Tobacco BY-2 Cells in a 3D Volume by Two-Photon Spinning-Disk Confocal Microscopy.
Journal Methods Mol Biol
Abstract Plant cells have a unique organization of microtubules during mitosis and cytokinesis. However, it is difficult to observe cell division in plant cells because of the distortion of images caused by thick samples. We addressed this issue by two-photon excitation coupled with spinning-disk confocal microscopy. Herein, we describe the method for observing microtubules and nuclei during cell division in tobacco BY-2 cells by two-photon spinning-disk confocal microscopy. The protocol includes the transformation of BY-2 cells with a plasmid for the expression of tubulin and histone markers, preparation of cells for microscopy, and operation of the two-photon spinning-disk microscope.
Volume 2872
Pages 37-50
Published 2025-1-1
DOI 10.1007/978-1-0716-4224-5_3
PMID 39616567
MeSH Cell Division* Cell Nucleus / metabolism Imaging, Three-Dimensional / methods Microscopy, Confocal* / methods Microtubules* / metabolism Nicotiana* / cytology Nicotiana* / genetics Nicotiana* / metabolism Plant Cells / metabolism Tubulin / genetics Tubulin / metabolism
Resource
Arabidopsis / Cultured plant cells, genes rpc00001