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*CIL – Cell Image Library accession number. Please use this to reference an image.

CIL:23605*  Cite 
Description

Timelapse movie of GFP-labeled microtubules in kar9delta cells (arrested in S-phase) combined with a deletion of the CAP-Gly domain of Nip100. The number of cells in which the spindle moved from the mother cell compartment through the neck was decreased in the CAP-Gly mutant. However, in CAP-Gly mutant cells where the spindle did move through the neck, the frequency of transits of the spindle, back-and-forth through the neck was similar to wild-type cells (compare with Movies S1 and S3, CIL#s 23599, 23604). Movie plays at 50x real time and is Movie S4 in Proc Natl Acad Sci. 2009. 106: 5147-5152.

Technical Details

S. cerevisiae (MATa NIP100deltaCAP-Gly DYN1-3GFP::TRP1 kar9delta::hygB ura3-52 lys2-801 leu2-delta1::GFP-TUB1::LEU2 his3-delta200 trp1-delta63) expressing GFP-tubulin (GFP-TUB1) and dynein heavy chain tagged with 3GFP (DYN1-3GFP) were arrested in hydroxyurea and mounted on agarose pads for microscopy. Images were captured on an Olympus Bmax-60F microscope equipped with a 1.35NA 100× UPlanApo objective, spinning disc Confocal Scanner Unit (CSU10), Picarro Cyan laser (488 nm), and a Stanford Photonics XR-Mega10 ICCD camera, by using QED software (Media Cybernetics). Image analysis was performed by using ImageJ. Timelapse images were captured at 10-s intervals for 15 min and each image in the movie represents a composite of 9 planes separated by 500 nm.

Biological Sources
NCBI Organism Classification
Saccharomyces cerevisiae S288c
Cell Line
NIP100deltaCAP-Gly, kar9delta
Cellular Component
spindle pole body
cytoplasmic microtubule
microtubule
dynein complex
dynactin complex
spindle pole
cell cortex
Attribution
Names
Jeffrey K. Moore
David Sept
John A. Cooper
Published
Proc Natl Acad Sci. 2009. 106: 5147-5152.
Pubmed
19279216
Citation
Digital Object Identifier (DOI)
doi:10.7295/W9CIL23605
Archival Resource Key (ARK)
ark:/b7295/w9cil23605
Grouping This image is part of a group.
Imaging
Image Type
recorded image
Image Mode
spinning disk confocal microscopy
time lapse microscopy
Parameters Imaged
fluorescence emission
Source of Contrast
distribution of a specific protein
Visualization Methods
EGFP
Processing History
each frame is a composite of 9 planes
Data Qualifiers
processed data
Sample Preparation
Methods
living tissue
Relation To Intact Cell
whole mounted tissue
Dimensions
Spatial Axis Image Size Pixel Size
X 320px ——
Y 250px ——
Time 10 seconds 97