scholarly journals Microtubule organization within mitotic spindles revealed by serial block face scanning EM and image analysis

2016 ◽  
Author(s):  
Faye M. Nixon ◽  
Thomas R. Honnor ◽  
Georgina P. Starling ◽  
Alison J. Beckett ◽  
Adam M. Johansen ◽  
...  
Keyword(s):  
Mitotic Spindle ◽  
Spatial Arrangement ◽  
Mitotic Spindles ◽  
Resolution Limit ◽  
Microtubule Organization ◽  
Computational Tools ◽  
Face Scanning ◽  
Block Face ◽  
Bridging Fibers ◽  
Scanning Em

AbstractSerial block face scanning electron microscopy (SBF-SEM) is a powerful method to analyze cells in 3D. Here, working at the resolution limit of the method, we describe a correlative light-SBF-SEM workflow to resolve microtubules of the mitotic spindle in human cells. We present three examples of uses for this workflow which are not practical by light microscopy and/or TEM. First, distinguishing closely associated microtubules within K-fibers; second, resolving bridging fibers in the mitotic spindle; third, visualizing membranes in mitotic cells, relative to the spindle apparatus. Our workflow also includes new computational tools for exploring the spatial arrangement of MTs within the mitotic spindle. We use these tools to show that microtubule order in mitotic spindles is sensitive to the level of TACC3 on the spindle.

scholarly journals Microtubule organization within mitotic spindles revealed by serial block face scanning electron microscopy and image analysis

2017 ◽  
Vol 130 (10) ◽  
pp. 1845-1855 ◽  
Author(s):  
Faye M. Nixon ◽  
Thomas R. Honnor ◽  
Nicholas I. Clarke ◽  
Georgina P. Starling ◽  
Alison J. Beckett ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Image Analysis ◽  
Mitotic Spindles ◽  
Microtubule Organization ◽  
Face Scanning ◽  
Block Face ◽  
Scanning Electron

3-D reconstruction and analysis of mammalian mitotic spindle structure

1992 ◽  
Vol 50 (1) ◽  
pp. 578-579
Author(s):  
Kent McDonald ◽  
David Mastronarde ◽  
Rubai Ding ◽  
Eileen O'Toole ◽  
J. Richard McIntosh
Keyword(s):  
Cross Sections ◽  
Mitotic Spindle ◽  
Experimental Studies ◽  
Video Camera ◽  
Three Dimensions ◽  
Mitotic Spindles ◽  
Black And White ◽  
Reconstruction Methods ◽  
Spindle Structure ◽  
Tissue Culture Line

Mammalian spindles are generally large and may contain over a thousand microtubules (MTs). For this reason they are difficult to reconstruct in three dimensions and many researchers have chosen to study the smaller and simpler spindles of lower eukaryotes. Nevertheless, the mammalian spindle is used for many experimental studies and it would be useful to know its detailed structure.We have been using serial cross sections and computer reconstruction methods to analyze MT distributions in mitotic spindles of PtK cells, a mammalian tissue culture line. Images from EM negatives are digtized on a light box by a Dage MTI video camera containing a black and white Saticon tube. The signal is digitized by a Parallax 1280 graphics device in a MicroVax III computer. Microtubules are digitized at a magnification such that each is 10-12 pixels in diameter.

Laser Scanning Confocal Microscopy and Three-Dimesnsional Reconstruction of the Mitotic Spindles of Unequally Dividing Embryonic Cells

1990 ◽  
Vol 48 (3) ◽  
pp. 166-167
Author(s):  
J. Holy ◽  
G. Schatten
Keyword(s):  
Confocal Microscopy ◽  
Mitotic Spindle ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Confocal Microscopy ◽  
Two Dimensions ◽  
Embryonic Cells ◽  
Mitotic Spindles ◽  
Cleavage Division ◽  
Scanning Confocal Microscopy

One of the classic limitations of light microscopy has been the fact that three dimensional biological events could only be visualized in two dimensions. Recently, this shortcoming has been overcome by combining the technologies of laser scanning confocal microscopy (LSCM) and computer processing of microscopical data by volume rendering methods. We have employed these techniques to examine morphogenetic events characterizing early development of sea urchin embryos. Specifically, the fourth cleavage division was examined because it is at this point that the first morphological signs of cell differentiation appear, manifested in the production of macromeres and micromeres by unequally dividing vegetal blastomeres.The mitotic spindle within vegetal blastomeres undergoing unequal cleavage are highly polarized and develop specialized, flattened asters toward the micromere pole. In order to reconstruct the three-dimensional features of these spindles, both isolated spindles and intact, extracted embryos were fluorescently labeled with antibodies directed against either centrosomes or tubulin.

scholarly journals Compact-sized Cutting System for a Serial-block-face Scanning Electron Microscopy

2021 ◽  
Vol 27 (S1) ◽  
pp. 3176-3177
Author(s):  
Nanami Takagi ◽  
Norio Yamashita ◽  
Yuki Tsujimura ◽  
Hiroshi Takemura ◽  
Sze Keat Chee ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Face Scanning ◽  
Block Face ◽  
Cutting System ◽  
Scanning Electron

scholarly journals Plasma Cleaning Improves the Image Quality of Serial Block-face Scanning Electron Microscopy (SBFSEM) Volumetric Data Sets

2017 ◽  
Vol 23 (S1) ◽  
pp. 1266-1267 ◽  
Author(s):  
Barbara Armbruster ◽  
Christopher Booth ◽  
Stuart Searle ◽  
Michael Cable ◽  
Ronald Vane
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Image Quality ◽  
Data Sets ◽  
Plasma Cleaning ◽  
Volumetric Data ◽  
Face Scanning ◽  
Block Face ◽  
Scanning Electron
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